Atlas of Genetics and Cytogenetics

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Scope

The Atlas of Genetics and Cytogenetics in Oncology and Haematology is a peer reviewed on-line journal in open access, devoted to genes, cytogenetics, and clinical entities in cancer, and cancer-prone diseases. It presents structured review articles ("cards") on genes, leukaemias, solid tumours, cancer-prone diseases, more traditional review articles on these and also on surrounding topics ("deep insights"), case reports in hematology, and educational items in the various related topics for students in Medicine and in Sciences.

Editorial correspondance

Jean-Loup Huret Genetics, Department of Medical Information, University Hospital F-86021 Poitiers, France tel +33 5 49 44 45 46 or +33 5 49 45 47 67 [email protected] or [email protected]

Staff Mohammad Ahmad, Mélanie Arsaban, Houa Delabrousse, Marie-Christine Jacquemot-Perbal, Maureen Labarussias, Vanessa Le Berre, Anne Malo, Catherine Morel-Pair, Laurent Rassinoux, Sylvie Yau Chun Wan - Senon, Alain Zasadzinski. Philippe Dessen is the Database Director, and Alain Bernheim the Chairman of the on-line version (Gustave Roussy Institute – Villejuif – France).

The Atlas of Genetics and Cytogenetics in Oncology and Haematology (ISSN 1768-3262) is published 12 times a year by ARMGHM, a non profit organisation, and by the INstitute for Scientific and Technical Information of the French National Center for Scientific Research (INIST-CNRS) since 2008.

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The PDF version of the Atlas of Genetics and Cytogenetics in Oncology and Haematology is a reissue of the original articles published in collaboration with the Institute for Scientific and Technical Information (INstitut de l’Information Scientifique et Technique - INIST) of the French National Center for Scientific Research (CNRS) on its electronic publishing platform I-Revues. Online and PDF versions of the Atlas of Genetics and Cytogenetics in Oncology and Haematology are hosted by INIST-CNRS. Atlas of Genetics and Cytogenetics

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Editor

Jean-Loup Huret (Poitiers, France) Editorial Board

Sreeparna Banerjee (Ankara, Turkey) Solid Tumours Section Alessandro Beghini (Milan, Italy) Genes Section Anne von Bergh (Rotterdam, The Netherlands) Genes / Leukaemia Sections Judith Bovée (Leiden, The Netherlands) Solid Tumours Section Vasantha Brito-Babapulle (London, UK) Leukaemia Section Charles Buys (Groningen, The Netherlands) Deep Insights Section Anne Marie Capodano (Marseille, France) Solid Tumours Section Fei Chen (Morgantown, West Virginia) Genes / Deep Insights Sections Antonio Cuneo (Ferrara, Italy) Leukaemia Section Paola Dal Cin (Boston, Massachussetts) Genes / Solid Tumours Section Louis Dallaire (Montreal, Canada) Education Section Brigitte Debuire (Villejuif, France) Deep Insights Section François Desangles (Paris, France) Leukaemia / Solid Tumours Sections Enric Domingo-Villanueva (London, UK) Solid Tumours Section Ayse Erson (Ankara, Turkey) Solid Tumours Section Richard Gatti (Los Angeles, California) Cancer-Prone Diseases / Deep Insights Sections Ad Geurts van Kessel (Nijmegen, The Netherlands) Cancer-Prone Diseases Section Oskar Haas (Vienna, Austria) Genes / Leukaemia Sections Anne Hagemeijer (Leuven, Belgium) Deep Insights Section Nyla Heerema (Colombus, Ohio) Leukaemia Section Jim Heighway (Liverpool, UK) Genes / Deep Insights Sections Sakari Knuutila (Helsinki, Finland) Deep Insights Section Lidia Larizza (Milano, Italy) Solid Tumours Section Lisa Lee-Jones (Newcastle, UK) Solid Tumours Section Edmond Ma (Hong Kong, China) Leukaemia Section Roderick McLeod (Braunschweig, Germany) Deep Insights / Education Sections Cristina Mecucci (Perugia, Italy) Genes / Leukaemia Sections Yasmin Mehraein (Homburg, Germany) Cancer-Prone Diseases Section Fredrik Mertens (Lund, Sweden) Solid Tumours Section Konstantin Miller (Hannover, Germany) Education Section Felix Mitelman (Lund, Sweden) Deep Insights Section Hossain Mossafa (Cergy Pontoise, France) Leukaemia Section Stefan Nagel (Braunschweig, Germany) Deep Insights / Education Sections Florence Pedeutour (Nice, France) Genes / Solid Tumours Sections Elizabeth Petty (Ann Harbor, Michigan) Deep Insights Section Susana Raimondi (Memphis, Tennesse) Genes / Leukaemia Section Mariano Rocchi (Bari, Italy) Genes Section Alain Sarasin (Villejuif, France) Cancer-Prone Diseases Section Albert Schinzel (Schwerzenbach, Switzerland) Education Section Clelia Storlazzi (Bari, Italy) Genes Section Sabine Strehl (Vienna, Austria) Genes / Leukaemia Sections Nancy Uhrhammer (Clermont Ferrand, France) Genes / Cancer-Prone Diseases Sections Dan Van Dyke (Rochester, Minnesota) Education Section Roberta Vanni (Montserrato, Italy) Solid Tumours Section Franck Viguié (Paris, France) Leukaemia Section José Luis Vizmanos (Pamplona, Spain) Leukaemia Section Thomas Wan (Hong Kong, China) Genes / Leukaemia Sections

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) Atlas of Genetics and Cytogenetics

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Volume 14, Number 4, April 2010

Table of contents

Gene Section

ADIPOR1 (adiponectin receptor 1) 334 Virginia Kaklamani, Christos Mantzoros AKT1 (v-akt murine thymoma viral oncogene homolog 1) 336 Daniela Etro, Silvia Missiroli, Francesca Buontempo, Luca Maria Neri, Silvano Capitani B3GNT6 (UDP-GlcNAc:betaGal beta-1,3-N-acetylglucosaminyltransferase 6 (core 3 synthase)) 353 Neeru M Sharma, Prakash Radhakrishnan, Shuhua Tan, Pi-Wan Cheng BAX (BCL2-associated X protein) 356 Hellinida Thomadaki, Andreas Scorilas CEACAM1 (carcinoembryonic antigen-related cell adhesion molecule 1 (biliary glycoprotein)) 361 Yasunobu Matsuda DAB2 (disabled homolog 2, mitogen-responsive phosphoprotein (Drosophila)) 365 Maurizio Orlandini DLG1 (discs, large homolog 1 (Drosophila)) 368 Paola Massimi, Lawrence Banks EEF1A1 (eukaryotic translation elongation factor 1 alpha 1) 377 Bruna Scaggiante, Giorgio Manzini FHL2 (four and a half LIM domains 2) 383 Marie Lin, William Cheung ING2 (inhibitor of growth family, member 2) 386 Susanne Jennek, Aria Baniahmad KLK7 (kallikrein-related peptidase 7) 389 Ying Dong, John Lai, Judith A Clements LATS1 (LATS, large tumor suppressor, homolog 1 (Drosophila)) 397 Stacy Visser, Xiaolong Yang NOL3 (nucleolar protein 3 (apoptosis repressor with CARD domain)) 400 Gloria Kung, Wendy McKimpson, Richard N Kitsis NPY (neuropeptide Y) 404 Massimiliano Ruscica, Elena Dozio, Paolo Magni ORAOV1 (oral cancer overexpressed 1) 409 Lu Jiang, Jinsheng Yu, Qianming Chen PAX8 (paired box 8) 412 Dario de Biase, Luca Morandi, Giovanni Tallini PTGIS (prostaglandin I2 (prostacyclin) synthase) 418 Inês Cebola, Miguel A Peinado RASL11B (RAS-like, family 11, member B) 421 Stefan Lorkowski

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) Atlast(11;14)(q13;q32) of Genetics in multiple myeloma and Cytogenetics Huret JL, Laï JL in Oncology and Haematology OPEN ACCESS JOURNAL AT INIST-CNRS

TJP2 (tight junction protein 2 (zona occludens 2)) 423 Lorenza Gonzalez-Mariscal, Erika Garay, Miguel Quiros, Rocio Tapia

Leukaemia Section

+16 or trisomy 16 (solely) 429 Jean-Loup Huret t(3;3)(q27;q29) 431 Jean-Loup Huret t(8;9)(q24;q13) 433 Jean-Loup Huret

Solid Tumour Section t(2;22)(q31;q12) in a small round cell tumour 434 Jean-Loup Huret

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) Atlas of Genetics and Cytogenetics

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Gene Section Mini Review

ADIPOR1 (adiponectin receptor 1) Virginia Kaklamani, Christos Mantzoros Department of Medicine, Division of Hematology/Oncology, Northwestern University, Feinberg School of Medicine, Chicago, IL, 676 N St Clair St. Suite 850, Chicago, IL 60611, USA (VK); Department of Medicine, Division of Endocrinology, Harvard University Medical School, Beth Israel Deaconess Medical Center, E/St 816, 330 Brookline Ave, Boston, MA02215, USA (CM)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/ADIPOR1ID44512ch1q32.html DOI: 10.4267/2042/44724 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Protein Other names: ACDCR1; CGI-45; CGI45; FLJ25385 Description FLJ42464; PAQR1; TESBP1A Receptor for globular and full-length adiponectin HGNC (Hugo): ADIPOR1 (APM1), an essential hormone secreted by adipo-cytes Location: 1q32.1 that counteracts the effects of insulin. DNA/RNA Expression Widely expressed. Highly expressed in adipose tissue Description and skeletal muscle. Expressed at intermed-iate level in Eight exons spanning 17.5 kb. Transcription is from brain, heart, spleen, kidney, liver, placenta, lung and telomere to centromere. peripheral blood leukocytes. Weakly expressed in colon, thymus and small intestine. We have also shown Transcription expression in normal and cancerous breast tissue as Transcription produces 11 different mRNAs, 10 well as in other cancers such as prostate, colon, alternatively spliced variants and 1 unspliced form. endometrial. It is also present in several cell lines. There are 6 probable alternative promoters, 2 non overlapping alternative last exons and 8 validated Localisation alternative polyadenylation sites. The mRNAs appear ADIPOR1 localizes to the plasma membrane. to differ by truncation of the 5' end, truncation of the 3' end, presence or absence of 6 cassette exons, Function overlapping exons with different boundaries. 9 spliced Receptor for globular and full-length adiponectin and the unspliced mRNAs putatively encode good (APM1), an essential hormone secreted by adipo-cytes proteins, altogether 9 different isoforms (3 complete, 6 that acts as an antidiabetic. Probably involved in partial), some containing Hly-III related proteins metabolic pathways that regulate lipid meta-bolism domain (Pfam) some transmembrane domains (Psort2); such as fatty acid oxidation. Mediates increased 1 of the 3 complete proteins appears to be secreted. The AMPK, PPARa ligand activity, fatty acid oxidation and remaining mRNA variant (spliced) appears not to glucose uptake by adiponectin. Has some high-affinity encode a good protein. for globular adiponectin but low-affinity for full-length adiponectin.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 334 ADIPOR1 (adiponectin receptor 1) Kaklamani V, Mantzoros C

Genomic structure of ADIPOR1. Blue boxes indicate exons.

Homology Prostate Cancer The mouse, human, and rat Adipor1 protein Note (ADIPOR1) contains 375 amino acids. Human and Adiponectin levels have been shown to correlate with mouse ADIPOR1 share 96.8% identity. prostate cancer risk (Barb et al., 2007; Arisan et al., 2009). Mutations References Note One functional polymorphism has been described. Petridou E, Mantzoros C, Dessypris N, Koukoulomatis P, Addy Rs7539542 has been found to modulate expression of C, Voulgaris Z, Chrousos G, Trichopoulos D. Plasma adiponectin concentrations in relation to endometrial cancer: a ADIPOR1 mRNA (Soccio et al., 2006). case-control study in Greece. J Clin Endocrinol Metab. 2003 Mar;88(3):993-7 Implicated in Dal Maso L, Augustin LS, Karalis A, Talamini R, Franceschi S, Trichopoulos D, Mantzoros CS, La Vecchia C. Circulating Breast Cancer adiponectin and endometrial cancer risk. J Clin Endocrinol Note Metab. 2004 Mar;89(3):1160-3 Adiponectin has been implicated in breast cancer. The Dieudonne MN, Bussiere M, Dos Santos E, Leneveu MC, breast cancer cell lines MCF-7, MDB-MB-231 and Giudicelli Y, Pecquery R. Adiponectin mediates antiproliferative and apoptotic responses in human MCF7 breast cancer cells. T47D were found to express both adiponectin receptors Biochem Biophys Res Commun. 2006 Jun 23;345(1):271-9 ADIPOR1/ADIPOR2 (Dieudonne et al., 2006; Korner et al., 2007) and exposure of T47D cells to adiponectin, Soccio T, Zhang YY, Bacci S, Mlynarski W, Placha G, Raggio G, Di Paola R, Marucci A, Johnstone MT, Gervino EV, significantly inhibited their proliferation (Korner et al., Abumrad NA, Klein S, Trischitta V, Doria A. Common 2007). rs2232853 CT genotype (OR=1.67; 95% CI haplotypes at the adiponectin receptor 1 (ADIPOR1) locus are 1.23-2.26) and the combination of rs7539542 GC associated with increased risk of coronary artery disease in (OR=0.59; 95% CI 0.36-0.98) and CC genotypes type 2 diabetes. Diabetes. 2006 Oct;55(10):2763-70 (OR=0.57; 95% CI 0.35-0.94) were significantly Barb D, Williams CJ, Neuwirth AK, Mantzoros CS. Adiponectin associated with breast cancer risk. The high expressing in relation to malignancies: a review of existing basic research rs2241766 G allele (GG and GT genotypes) was and clinical evidence. Am J Clin Nutr. 2007 Sep;86(3):s858-66 associated with decreased breast cancer risk (OR=0.64; Körner A, Pazaitou-Panayiotou K, Kelesidis T, Kelesidis I, 95% CI 0.49-0.83). The low expressing rs1501299 G Williams CJ, Kaprara A, Bullen J, Neuwirth A, Tseleni S, Mitsiades N, Kiess W, Mantzoros CS. Total and high- allele was associated with increased breast cancer risk: molecular-weight adiponectin in breast cancer: in vitro and in for TG: OR=1.59; 95% CI 1.03-2.48, for GG: OR= vivo studies. J Clin Endocrinol Metab. 2007 Mar;92(3):1041-8 1.80; 95% CI 1.14-2.85. Kaklamani VG, Wisinski KB, Sadim M, Gulden C, Do A, Offit K, Colon Cancer Baron JA, Ahsan H, Mantzoros C, Pasche B. Variants of the adiponectin (ADIPOQ) and adiponectin receptor 1 (ADIPOR1) Note genes and colorectal cancer risk. JAMA. 2008 Oct Adiponectin has also been implicated in colon cancer 1;300(13):1523-31 risk. rs266729 polymorphism was signi-ficantly Arisan ED, Arisan S, Atis G, Palavan-Unsal N, Ergenekon E. associated with colon cancer risk: the RR for the Serum adipocytokine levels in prostate cancer patients. Urol GG/CG genotypes was rs266729 was associated with a Int. 2009;82(2):203-8 reduced colorectal cancer risk: O.R. 0.73, 95% CI This article should be referenced as such: (0.53-0.99) (Kaklamani et al., 2008). Kaklamani V, Mantzoros C. ADIPOR1 (adiponectin receptor 1). Endometrial Cancer Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4):334-335. Note Adiponectin levels have been shown to correlate with endometrial cancer risk (Petridou et al., 2003; Dal et al., 2004).

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 335 Atlas of Genetics and Cytogenetics

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Gene Section Review

AKT1 (v-akt murine thymoma viral oncogene homolog 1) Daniela Etro, Silvia Missiroli, Francesca Buontempo, Luca Maria Neri, Silvano Capitani Department of Morphology and Embryology, Human Anatomy Section, Ferrara University, 44100 Ferrara, Italy (DE, SM, FB, LMN, SC)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/AKT1ID355ch14q32.html DOI: 10.4267/2042/44725 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Transcription The human AKT1 coding sequence consists of 1443 bp Other names: AKT; C-AKT; EC 2.7.11.1; from the start codon to the stop codon. Multiple MGC99656; PKB; PKB-ALPHA; PRKBA; RAC; alternatively spliced transcript variants have been found RAC-ALPHA; RAC-PK-alpha for this gene (Entrez Gene). HGNC (Hugo): AKT1 Pseudogene Location: 14q32.33 No pseudogene of AKT1 known. Note Location in the mouse: chromosome 12, 57.0 cM, Protein 113892032 to 113912401 bp, complement strand. For a comparison of the gene location among Homo Note sapiens, mouse and rat see: NCBI Map Viewer. Although the AKT isoforms are activated in a similar manner and share the same downstream substrates, DNA/RNA indicating functional redundancy of the AKT isoforms, their biological function is likely to be different in Description AKT-knockout mouse models. AKT1 mutant mice display developmental defects, showing decreased size The human AKT1 gene is composed of 14 exons in all organs and impaired placental development spanning a genomic region of about 26.4 Kb. The open (Yang et al., 2004). reading frame of the coding region is 1443 bp.

a. Genomic organization of human AKT1. The line indicates untranslated regions and boxes indicate coding regions (exon 1-14) of the gene. Exon and intron lengths (in bp) are reported in the upper and lower part of the diagram, respectively. The ATG transcription start site is located in exon 2 and the TGA termination codon is located in exon 14. b. mRNA of human AKT1.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 336 AKT1 (v-akt murine thymoma viral oncogene homolog 1) Etro D, et al.

a. Diagram of the human AKT1 protein in scale. The protein domains and their length (indicated by number of limiting residues) are reported. AKT1 contains a pleckstrin homology domain (PH), an helical region (Helix), a kinase domain (Kinase), and a regulatory motif (Regulatory). The two phosphorylation sites essential for complete activation of AKT1 (threonine 308, serine 473) are indicated in the diagram. C: carboxyl-terminal; N: amino-terminal. b. Schematic representation of the AKT signaling activation and regulation.

AKT1 deficient mice exhibit perinatal morbidity with involved as cytoskeletal constituents or in intracellular partial lethality between E13.5 and 3 weeks after birth signaling; the structure of the PH domain consists of and growth retardation. Surviving adults are fertile, but two perpendicular anti-parallel beta-sheets followed by show 20% weight reduction accom-panied by reduced a C-terminal amphipathic helix; the common fold of sizes of multiple organs, and enhanced apoptosis in PH domains is electrostatically polarized. The PH some cell types. No effect seen on glucose metabolism. domain recruits AKT to the plasma membrane by Moreover, AKT1/ AKT2 double-knockout mice phosphoinositides binding and is required for display impeded adipogenesis, severe growth activation. deficiency including impaired skin development, severe The kinase domain has been evolutionarily conserved muscle atrophy, impaired bone development and die from Escherichia coli to Homo sapiens; conserved shortly after birth (Peng et al., 2003). regions are: i) a glycine-rich stretch of residues in close Description proximity of a lysine amino acid (179, by similarity), involved in ATP binding; ii) an highly conserved Structure. AKT1 protein consists of 480 amino acids, activation loop, called T-loop, located between DFG with a molecular weight of 55,686 Da. AKT1 is and APE motifs, with a threonine residue important for constituted by a PH domain, a short helical region, a enzyme activation; iii) a conserved aspartic acid (274, catalytic kinase domain and a regulatory hydrophobic by similarity) as proton acceptor residue, important for motif. the catalytic activity of the enzyme. The kinase domain PH domain is a conserved domain of about 100 catalyzes the transfer of the gamma-phosphoryl group residues that occurs in a wide range of proteins from ATP to serine/threonine residues on a consensus

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 337 AKT1 (v-akt murine thymoma viral oncogene homolog 1) Etro D, et al.

sequence on protein substrates, resulting in a Beside these essential activation sites, threonine 72 and conformational change affecting protein function, serine 246 residues undergo auto-phosphoryla-tion (Li cellular location or association with other proteins et al., 2006), serine 124 and threonine 450 residues are (Knighton et al., 1991). constitutively phosphorylated, while tyrosine 315 and The carboxyl-terminal hydrophobic regulatory domain 326 in the activation loop can be phosphorylated by Src contains several proline-rich regions that potentially kinase, maybe regulating AKT1 activity (Chen et al., serve as protein-protein interaction sites with important 2001). roles in regulation of AKT1 activity; this region Regulation. AKT activation is inversely regulated by contains the 473 residue important for the activation phosphatases: PH domain leucine-rich repeat protein process. This domain possesses the F-X-X-F/Y-S/T- phosphatase (PHLPP) dephosphorylates the serine 473 Y/F hydrophobic motif, where X is any amino acid, residue of AKT1 (Brognard et al., 2007), and protein that is characteristic of the AGC kinase family; in phosphatase 2 (PP2) dephosphorylates the threonine mammalian AKT isoforms, this motif is identical 308 residue (Gao et al., 2005). PI(3,4,5)P3 is (FPQFSY) and is thought to be very important for the hydrolyzed by phosphatase and tensin homolog deleted enzymatic activity. The conserved SH3-domain binding on chromosome 10 (PTEN) and Src homology domain- motif P-X-X-P in the regulatory region is involved in containing inositol phosphatases SHIP1/SHIP2. PTEN the interaction between AKT1 and its upstream antagonizes PI3K activity by removing the phosphate tyrosine kinase Src (Jiang et al., 2003). at the D3 position generating PI(4,5)P2 (Maehama et The crystallographic structure of AKT1 has been al., 1998), while SHIP1/2 dephosphorylates the D5 solved (PDB ID 3CQW, 3CQU). position to produce PI(3,4)P2 (Deleris et al., 2003; Damen et al., 1996). Activation. The serine-threonine protein kinase AKT1 is a catalytically inactive cytoplasmic protein. AKT Expression activation occurs by means of stimulation of the growth AKT1 is the predominant isoform in the major part of factor receptor-associated phosphatidylinositol 3-kinase tissues as determined by using quantitative RT-PCR (PI3K) and is a multi-step process that involves both (Yang et al., 2003) and is ubiquitously expressed in membrane translocation and phosphorylation. When most tissues at high levels and in all the human cell PI3K is activated by either growth factors, cytokines or types so far analyzed (Hanada et al., 2004; Zinda et al., hormones, PI3K generates 3'-phosphorylated 2001). A Northern blot analysis of AKT1 in rat tissues phosphoinositides, i.e. phosphatidylinositol-3,4,5- indicated lower expression levels in kidney, liver, and trisphosphate (PIP 3) and phosphatidylinositol-3,4- spleen (Coffer et al., 1991). bisphosphate (PIP 2) at the plasma membrane. Both phospholipids bind with high affinity to the PH Localisation domain, mediating membrane translocation of AKT. At AKT1 protein is predominantly cytoplasmic; it has the membrane, AKT1 is phosphorylated at threonine been found at the plasma membrane for its activation 308 by PDK1 (Andjelkovic et al., 1997; Walker et al., and activated AKT1 is able to translocate into the 1998) and at serine 473 by a second kinase identified nucleus. AKT1 translocation into nucleus has been with mTOR when bound to Rictor in the so called demonstrated in several cell lines in response to stimuli TORC2 complex (Santos et al., 2001; Sarbassov et al., as after IGF-I treatment of NIH3T3 cells (Meier, 1997), 2005); however, it is still controversial if this second NGF stimulation of PC12 cells (Xuan Nguyen et al., phosphorylation may occur by DNA-dependent protein 2006; Borgatti et al., 2003), EPO in K562 cells and kinase (Feng et al., 2004; Hill et al., 2002). Other IGF-I or PDGF mitogen factors in MC3T3 (Neri et al., kinases that have been reported to phosphorylate serine 2002). Also if AKT1 contains a sequence for nuclear 473 are PKC (Kawakami et al., 2004), integrin-linked export rich in leucine (Saji et al., 2005) and some kinase (ILK) (Troussard et al., 2003; Lynch et al., proteins may have a role of localization signal for its 1999; Delcommenne et al., 1998), MAP kinase- intranuclear migration, a nuclear localisation sequence activated protein kinase-2 (MK2) (Rane et al., 2001), on AKT1 inside motif has not yet been identified. PDK-1 (Balendran et al., 1999) or Akt itself (Toker et Function al., 2000). The full activation of AKT1 requires phosphorylation at both sites; threonine 308 AKT mediates many of the downstream events of the phosphorylation increases the enzymatic activity up to PI3K signal transduction pathway by its serine- 100-fold and serine 473 phosphorylation by a further threonine kinase activity. AKT exhibits tight control 10-fold, thus both phosphorylation events enhance over cell viability and proliferation, having main role in AKT1 activity by 1000-fold (Kumar et al., 2005; Alessi apoptosis inhibition and promotion of cell cycle et al., 1996). The activation is rapid and specific, and it progression. AKT is involved also in differentiation; in is abrogated by mutations in the AKT PH domain. nervous system development AKT is a critical mediator Once activated, AKT1 dissociates from the membrane of growth factor-induced neuronal survival. Further, and phosphorylates targets in the cytoplasm and the cell AKT mediates glucose metabolism, angiogenesis, nucleus. translation, transcript-tional events, pre-mRNA splicing

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 338 AKT1 (v-akt murine thymoma viral oncogene homolog 1) Etro D, et al.

and other important nuclear functions such as regulates cell proliferation. AKT phospho-rylates chromatin condensation and genes transactivation. WNK1 on threonine 60 within the AKT consensus AKT exerts its kinase activity toward proteins sequence (Vitari et al., 2004). The neuro-fibromatosis-2 containing the minimal consensus sequence R/K-X- (NF2) tumour-suppressor gene encodes an intracellular R/K-X-X-S/T, where S or T are the phosphorylable membrane-associated protein, called merlin, with residues. More subtle AKT preferences were also growth-suppressive function. AKT phosphorylates uncovered for other residues surrounding the merlin on threonine 230 and serine 315 residues, phosphorylation site, such as a preference for T at -2 or abolishing binding partners and leading to merlin a bulky hydrophobic residue at +1 (Manning et al., degradation by ubiquitination (Tang et al., 2007). 2007). More than 400 different proteins contain-ing the Metabolism. AKT phosphorylates the GSK3alpha and consensus sequence for AKT phosphoryla-tion have GSK3beta isoforms, which are involved in metabolism been identified, also if many of them still have to be regulation by decreasing glycogen synthesis and characterized (Nicholson et al., 2002; Obenauer et al., increasing glycolytic enzymes transcription (Jope et al., 2003). The heterogeneity of proteins potentially 2004; Kohn et al., 1996), thus relating AKT activation phosphorylated by AKT supports the key role of this with high glycolysis efficiency in cancer cells kinase. Over 100 non-redundant AKT substrates are (Warburg effect). AKT1 is also involved in tolerance of reported in the literature, of which 25% do not contain cells to nutrient depletion, allowing tumor progression the minimal requirements for an AKT site. Around 40 under hypovascular conditions (Izuishi et al., 2000). substrates which mediate the pleiotropic AKT functions The TBC1 domain family member 1 (TBC1D1), AKT have been characterized (see table below). substrate phosphorylated on threonine 590, may be Apoptosis inhibition. Survival factors can suppress involved in controlling GLUT1 glucose transporter apoptosis and enhance survival of cells by activating expression through the mTOR/p70S6K pathway (Zhou AKT, which inactivates components of the apoptotic et al., 2008). The Rab-GAP AS160 (also known as machinery. AKT directly regulates apoptosis by TBC1D4) has emerged as an important direct target of phosphorylating and inactivating pro-apoptotic proteins AKT involved in GLUT4 trans-location to the plasma such as bad, which controls release of cytochrome c membrane (Sano et al., 2003). In hepatocytes, AKT can from mitochondria, caspase-9, which after AKT also inhibit gluconeogenesis and fatty acid oxidation dependent phospho-rylation promotes cell survival through direct phosphorylation on serine 570 of PGC- (Donepudi et al., 2002; Downward et al., 1999; Franke 1alpha (Li et al., 2007), which is a gene coactivator et al., 2003) and apoptosis signal-regulating kinase-1 with FoxO1 and other transcription factors. (ASK1), a mitogen-activated protein kinase involved in Angiogenesis. AKT plays important roles in stress- and cytokine-induced cell death that, once angiogenesis through effects in both endothelial cells phosphorylated on serine 83, reduces apoptosis (Autret and cells producing angiogenic signals. AKT activates et al., 2008; Datta et al., 1997; Del Peso et al., 1997; endothelial nitric oxide synthase (eNOS) through direct Zha et al., 1996). The pro-survival proline-rich AKT phosphorylation on the serine 1179 site, resulting in substrate of 40kDa (PRAS40) can be phosphorylated increased production of nitric oxide (NO) in vascular on threonine 246, attenuating its ability to inhibit endothelium, which stimulates vasodilatation, vascular mTORC1 kinase activity (Van der Haar, 2007). remodelling and angiogenesis (Iantorno et al., 2007). PRAS40 appears to protect neuronal cells from Translation. A well known AKT substrate is the apoptosis after stroke (Kovacina et al., 2003) and has serine/threonine kinase mammalian target of rapamycin been proposed to promote cell survival in cancer cells (mTOR), which controls the translation of several (Huang et al., 2005). proteins important for cell cycle progression and Proliferation. AKT can stimulate cell cycle growth (Starkman et al., 2005; Varma et al., 2007). progression through the inhibitory phosphorylation of AKT can directly phospho-rylate and activate mTOR, the cyclin-dependent kinase inhibitors p21 and p27 as well as cause indirect activation of mTOR by (Viglietto et al., 2002; Liang et al., 2002; Shin et al., phosphorylating two sites on the tuberous sclerosis 2002; Zhou et al., 2001; Rossig et al., 2001). The AKT complex 2 (TSC2) tumour suppressor protein, also dependent inhibition of GSK3 stimulates cell cycle called tuberin (Manning et al., 2002). mTOR forms two progression by stabilizing cyclin D1 expression (Diehl complexes: TORC1, in which mTOR is bound to et al., 1998). AKT activation can promote progression Raptor, and TORC2, in which mTOR is bound to through mitosis, even in the presence of DNA damage Rictor. In the TORC1 complex, mTOR signals to its (Kandel et al., 2002); a mechanism explaining this downstream effectors S6 kinase/ribosomal protein and observation is that AKT directly phosphorylates the 4EBP-1/eIF-4E to control protein translation. In the DNA damage checkpoint kinase Chk1 on serine 280 TORC2 complex, mTOR can phosphorylate AKT itself (King et al., 2004), blocking checkpoint function by thus providing a positive feedback on the pathway stimulating Chk1 translocation to the cytosol. With no (Sarbassov et al., 2005). The mTOR effector S6 kinase- K protein kinase-1 (WNK1) seems to be a negative 1 (S6K1) can also regulate the pathway by inhibiting regulatory element in the insulin signaling pathway that the insulin receptor substrate (IRS), thus preventing

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 339 AKT1 (v-akt murine thymoma viral oncogene homolog 1) Etro D, et al.

IRS proteins from activating the PI3K/AKT signaling subnuclear residency, cell proliferation, and mRNA (Harrington et al., 2004; Shah et al., 2004). The Y box- export activities through nuclear AKT dependent binding protein 1 (YB-1) is a DNA/RNA-binding phosphorylation on threonine 219 and phosphoino- protein through the Y-box motif in target sequences. sitide association (Okada et al., 2008). AKT AKT phosphorylates YB-1 on serine 102, leading to an specifically phosphorylates serine 350 of the Nur77 enhancement of cap-dependent translation of multidrug protein within its DNA-binding domain, decreasing its resistance 1 (MDR1) gene (Bader et al., 2008). transcriptional activity by 50-85% and connecting the Nuclear functions. Among the AKT substrates AKT axis with a nuclear receptor pathway (Pekarsky et identified into cell nucleus, acinus is a nuclear factor al., 2001). The breast cancer susceptibility gene required for chromatin condensation which induces BRCA1 encodes a nuclear phosphoprotein that acts as a resistance to caspases proteolysis and to apoptosis tumor suppressor; heregulin induces AKT-dependent when phosphorylated by AKT on serine 422 and 573 phosphorylation of BRCA1, which has been implicated (Hu et al., 2005). Phosphorylation of the murine double in altering its function (Altiok et al., 1999). minute 2 (MDM2/HDM2 in humans) oncogene by AKT promotes its translocation to the nucleus, where it negatively regulates p53 function with subsequent modification of the cell cycle in relation to DNA repair mechanisms (Vousden et al., 2002; Mayo et al., 2005). Several Akt substrates are nuclear transcription factors: AKT blocks forkhead trans-cription factors (FKHR/FOXO1) and in particular the FoxO subfamily- mediated transcription of genes that promote apoptosis, cell cycle arrest and metabolic processes. When phosphorylated by AKT, FKHR are sequestrated in the cytoplasm thus inhibiting transcription (Nicholson et al., 2002; Datta et al., 1997). AKT can phosphorylate IKK, indirectly increasing the activity of nuclear factor kappa B (NF-kB), which stimulates the trans-cription of pro-survival genes and regulates the immunity response (Ozes et al., 1999; Romashkova et al., 1999; Verdu et al., 1999). The cAMP-response element binding protein (CREB) is a direct target for phosphorylation by AKT, occurring on a site that increases binding of CREB to proteins necessary for induction of genes containing cAMP responsive elements (CREs) in their promoter regions; CREB has been shown to mediate AKT-induced expression of antiapoptotic genes bcl-2 and mcl-1 (Du et al., 1998). AKT can regulate the telomerase activity necessary for DNA replication; recombinant AKT was found to enhance telomerase activity by phosphorylating the human telomerase reverse transcriptase (hTERT) subunit, which contains a consensus motif as AKT substrate. The helix-loop-helix transcription factor tal1, required for blood cell development, is specifically phosphorylated by AKT at threonine 90, causing its nuclear redistribution (Palamarchuk et al., 2005b). Insulin induces GATA2 phosphorylation on serine 401 by AKT. GATA2 transcription factor is an inhibitor of adipogenesis and activator of vascular cells. AHNAK is * substrates assessed independently by multiple reports a protein of exceptionally large size localized into (Manning et al., 2007). nuclei and able to shuttle between nucleus and cytoplasm; it is downregulated in several tumors Homology (Amagai et al., 2004). It has been reported that in Homologs. AKT belongs to the AGC protein kinase epithelial cells its extranuclear localization is regulated family, sharing a high similarity in the catalytic domain by AKT dependent phosphorylation (Sussman et al., with more than 80 kinases from the AGC family 2001). ALY is a nuclear speckle protein implicated in (PhosphositePlus). Three isoforms, AKT1, AKT2 and mRNA export. The PI3K/AKT signaling regulates its AKT3, plus a fourth isoform defined AKTgamma1,

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 340 AKT1 (v-akt murine thymoma viral oncogene homolog 1) Etro D, et al.

have been identified in humans. They are codified by Somatic different genes with 80% sequence homology. Amplification and LOH. Amplification of AKT1 has The AKT isoforms share 80% homology in amino acid been described in human gastric adenocar-cinoma, in sequence. lung and other cancers (Staal, 1987; Lockwood et al., In particular, the identity between each domain of the 2008). AKT isoforms ranges from 76% to 84% in the PH High level amplification in breast tissues and LOH in domain, from 87% to 90% in the catalytic domain, and several tissues have been reported: CONAN: Copy from 66% to 76% in the C-terminal domain (Masure et Number Analysis. al., 1999; Kumar et al., 2005). The AKT isoforms are SNPs. 17 esonic variations (missense, synonymous and identical in the ATP binding region, except for one frameshift SNPs) have been described. residue: AKT1 A230 is conserved in AKT2 (A232), but switches in AKT3 (V228). Moreover, statistical significance for single markers and multilocus haplotypes has been reported for the Orthologs. AKT is evolutionarily conserved in association between the AKT1 gene variants in samples eukaryotes ranging from Caenorhabditis elegans to of families with schizophrenia using single-nucleotide man. The amino acid identity between C. elegans and polymorphisms (Schwab et al., 2005; Emamian et al., human AKT1 is around 60%; the mouse AKT1 is 90% 2004). homologous to human AKT1 at the nucleic acid level and 98% homologous at the amino acid level (Hanada Point mutation. The E17K mutation occurs in the et al., 2004; Bellacosa et al., 1993). lipid-binding pocket of AKT1 PH domain. Lysine 17 alters the electrostatic interactions of the pocket and For details see: HomoloGene. forms new hydrogen bonds with a phosphoinositide Also the phosphorylation sites on the AKT sub-strates ligand. This mutation activates AKT1 by means of are conserved amongst the orthologs from all pathological localization to the plasma membrane, mammals; this evolutionary conservation can be stimulates downstream signaling, transforms cells and indicative of the relevance of the substrate toward the induces leukemia in mice. The E17K mutation occurs AKT cellular functions. in a small percentage of human breast, ovarian, and colorectal cancers (Carpten et al., 2007). It has been found also in squamous cell carcinoma of the lung and in prostate cancer (Malanga et al., 2008; Boormans et al., 2008). Some authors suggested that this mutation may not play a crucial role in the development of the most types of human cancers (Kim et al., 2008). Implicated in Various cancers Mutations Prognosis Immunohistochemical analysis has been used to Note demonstrate prognostic significance of AKT1 Although mutation of AKT1 is rare, different types of activation. Phosphorylation of AKT1 at serine 473 has AKT1 alterations are involved in several human been associated with poor prognosis in cancer of the diseases, especially in cancer. skin (Dai et al., 2005), pancreas (Yamamoto et al., No AKT1 mutations have been collected in the 2004), liver (Nakanishi et al., 2005), prostate COSMIC database. (Kreisberg et al., 2004), breast (Perez-Tenorio et al., 2002), endometrium (Terakawa et al., 2003), stomach (Nam et al., 2003), brain (Ermoian et al., 2002) and blood (Min et al., 2004). It has been reported that AKT phosphorylation on both serine 473 and threonine 308 sites is a better predictor of poor prognosis in tumors versus normal tissues than serine 473 alone (Tsurutani Schematic representation of SNPs and point mutation in the AKT1 gene. Missense (red), synonymous (green) and et al., 2006; Kornblau et al., 2006). frameshift (blue) SNPs are indicated in the upper part; point Oncogenesis mutation is reported in the lower part of the figure. For details see: Single Nucleotide Polymorphism. The PI3K/AKT pathway is a prototypic survival signaling that is constitutively activated in many types Germinal of cancer, due to AKT gene amplification or as a result No germline mutations of AKT1 have been described. of mutations in components of the signaling that

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 341 AKT1 (v-akt murine thymoma viral oncogene homolog 1) Etro D, et al.

activates AKT. Once activated, signaling through AKT (Campbell et al., 2001). It has been shown that can be propagated to a diverse array of substrates. This activation of AKT/mTOR promotes angiogenesis via pathway is an attractive therapeutic target in cancer HIF1alpha stabilization in breast cancer cells because it serves as a convergence point for many (Laughner et al., 2001). Recent studies have shown that growth stimuli, and through its downstream substrates, AKT1 can attenuate breast cancer cell motility, controls cellular processes that contribute to cancer whereas AKT2 enhances this phenotype. AKT1 blocks progression. Moreover, activation of the PI3K/AKT the migration of breast cancer cells through GSK3beta pathway confers resistance to many types of cancer inactivation and transcription factor NFAT inhibition therapy, and is poor prognostic factor for several (Yoeli-Lerner et al., 2009). tumors. Thus, combining conventional therapy with Lung cancer PI3K/AKT pathway inhibitors can overcome this Note resistance. Although AKT1 mutations are apparently rare in Hyper-activation of AKT1 has been found associated to lung cancer (1.9%), the oncogenic properties of E17K- several human cancers: AKT1 may contribute to the development of a fraction -Thyroid carcinoma of lung carcinoma with squamous histotype (Malanga -Breast carcinoma et al., 2008). Adenocarcinomas of the lung commonly -Non-small cell lung carcinoma show an increase in the activity of PI3K/AKT signaling pathway. The simultaneous inhibition AKT1 siRNA -Gastric carcinoma and Bcl-xL function greatly enhanced the apoptotic -Gastro-intestinal stromal tumors response, suggesting that AKT1 and Bcl-xL control cell -Pancreatic carcinoma death in lung adenocarcinoma cells in a synergistic -Bile duct carcinoma manner (Qian et al., 2009). AKT1 is overexpressed as a direct result of gene amplification in lung cancer, -Ovarian carcinoma suggesting that amplification of this genome hotspot is -Prostate carcinoma a common mechanism of oncogene activation -Renal cell carcinoma (Lockwood et al., 2008). -Acute and chronic leukemia Gastric carcinoma -Multiple myeloma Note -Lymphoma AKT1 gene amplification has been observed in gastric Thyroid cancer carcinoma. Most gastric adenocarcinomas arise as a longterm complication of Helicobacter pylori infection Note of the stomach; phosphorylation of AKT and its Genetic alterations in the AKT pathway have been substrates is inducible by epithelial mitogens such as observed in anaplastic and follicular thyroid cancers, in EGF, which is implicated in the pathogenesis of H. particular AKT has been shown highly phosphorylated pylori gastritis (Ang et al., 2005). NF-KB activation in thyroid cancer cell lines and human thyroid cancer was frequently observed in early-stage gastric specimens (Liu et al., 2008; Mandal et al., 2005). carcinoma and was significantly correlated with better Activated AKT is common to both human and mouse prognosis and Akt activation (Lee et al., 2005). AKT follicular thyroid cancer and is correlated with activation and LOH of PTEN play an important role in increased cell motility in vitro and metastasis in vivo conferring a broad-spectrum chemoresistance in gastric (Kim et al., 2005). cancer patients (Oki et al., 2005). Breast cancer Colorectal cancer Note Note Somatic mutation E17K occurs in the PH domain of The transforming E17K point mutation in the PH AKT1 in 8% of human breast cancers (Carpten et al., domain of AKT1 in human colorectal cancer (6%) has 2007). Overexpression of cyclin D1 has been found in been identified (Carpten et al., 2007). The breast cancer; elevated cyclin D1 levels result in Src/PI3K/FAK/AKT pathway has been described as shortened cell cycle times and thereby contribute to responsible of colon cancer cells metastatic adhesion tumor progression. AKT is involved in this mechanism (Thamilselvan et al., 2007). Cytoplasmic by regulating cyclin D1 expression at transcription, mislocalization of p27, caused by activated AKT1, and translation and protein stability level (Nicholson et al., functional losses of p27 and p53 have been associated 2002). Anti-estrogens such as tamoxifen inhibit the with poor prognosis and are involved in the growth of (estrogens receptors) ER-positive breast development of various subtypes of colorectal cancer cancers by reducing the expression of estrogen- (Ogino et al., 2007). The inhibitor of the apoptosis regulated genes. AKT, by activating ER, protects breast protein (IAP) family member XIAP is essential for cell cancer cells from tamoxifen-induced apoptosis survival in colorectal cancer cells and is activated

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 342 AKT1 (v-akt murine thymoma viral oncogene homolog 1) Etro D, et al.

through the AKT pathway. The AKT-XIAP up- domain of AKT1 in human ovarian cancer (2%) has regulation was shown to be correlated to colorectal been identified (Carpten et al., 2007). The AKT cancer progression and may be a potential molecular pathway plays an important role in cell prolifera-tion, target for therapy (Takeuchi et al., 2005). migration, and invasion in ovarian cancer cells; Glioblastoma and gliosarcoma particular importance has the signaling specificity of AKT1, as the inhibition of AKT1 is sufficient to affect Note these events (Meng et al., 2006; Kim et al., 2008; Gu et AKT1 amplification and overexpression have been al., 2008). observed in human glioblastoma and gliosarcoma, a variant of glioblastoma multiforme characterized by Prostate cancer two components displaying gliomatous or sarcoma-tous Note differentiation (Actor et al., 2002; Staal et al., 1987). Increased AKT1 kinase activity was reported in more Glioblastomas frequently carry mutations in PTEN than 50% of prostate carcinomas. The androgen gene, which tumor suppressor properties are closely receptor (AR) factors phosphorylated by AKT lead to related to its inhibitory effect on the AKT signaling inhibition of their activity and blockade of androgen- (Knobbe et al., 2003). induced apoptosis in a prostate cancer cell line (Lin et al., 2001). A study of prostate cancer indicates that Pancreatic cancer AKT is involved more in cancer progression than Note initiation. The E17K mutation was identified in clinical It was reported that constitutively active AKT1 in prostate cancer samples. The mutation was mutually mouse pancreas requires S6 kinase 1 for insulinoma exclusive with respect to PTEN inactivation and PI3K formation (Alliouachene et al., 2008). AKT1 serine 473 activation; it was suggested that tumors carrying the may undergo both phosphorylation and O-GlcNAc AKT1 mutation may follow a more favourable clinical modification, and the balance between these events course (Boormans et al., 2008). may regulate murine beta-pancreatic cell apoptosis (Kang et al., 2008). All the AKT isoforms may have Renal cancer protective effects within the cell depending on the type Note of apoptotic stimuli in human pancreatic MiaPaCa-2 Phospho-AKT expression is significantly increased in cells (Han et al., 2008). Overexpression of bcl-2 is renal carcinoma cells. A decreased expression of PTEN common in pancreatic cancer, confers resistance to the may be an underlying mechanism for AKT activation apoptotic effect of chemo- and radiotherapy and is and thus an AKT inhibitor may be a therapeutic option accompanied to increased activity of AKT as well as its for the subset of renal cell carcinoma patients with downstream target IKK (Mortenson et al., 2007). elevated AKT activity (Hara et al., 2005). Hepatocellular carcinoma (HCC) Melanoma Note Note Hyper-activation of the AKT pathway frequently Common mutations and/or deregulated expression of occurs in HCC (Roberts et al., 2005). It was reported proteins of the AKT signaling, as B-RAF, PTEN, that Bortezomib induces apoptosis in HCC cell lines by MDM2 and AKT itself, were identified in melanoma down-regulating phospho-AKT. Down-regulation of (Ch'ng et al., 2009). AKT-dependent phosphorylation phospho-AKT may thus represent a biomarker for of hTERT increases telomerase activity in melanoma predicting clinical response to HCC treatment (Chen et cells, indicating that AKT promotes the al., 2008). Moreover, it was observed that a cancer immortalization of cancer cells by preventing stem cell population in HCC contributes to replicative senescence (Kang et al., 1999). chemoresistance through preferential activation of Acute leukemia AKT and bcl-2 cell survival response (Ma et al., 2008). Note Knockdown of insulin receptor substrate in primary The AKT signaling is important for governing cell human HCC HepG2 cell line resulted in reduction of survival and proliferation in acute myeloid leukemia insulin stimulated AKT1 phosphorylation at serine 473 (AML). The level of AKT phosphorylation on and 50% reduction in the basal level of phosphorylated threonine 308 but not on serine 473 is associated with mTOR (Ser 2448), indicating a pivotal role of the AKT high-risk cytogenetics and predicts poor overall signaling in HCC (Varma et al., 2008). It was also survival in AML (Gallay et al., 2009). AKT activation presented that AKT1 was upregulated in HCC cells, critically mediates survival during the early phase of and its active phosphorylated form was mainly located drug (i.e. imatinib) resistance development (Burchert et in the nucleus (Zhu et al., 2007). al., 2005). PTEN phosphorylation, associated with Ovarian cancer increased AKT phosphorylation, is found in 75% of AML (Cheong et al., 2003a). Also SHIP1 alteration is Note shown to result in AKT activation in AML cells (Luo et The transforming E17K point mutation in the PH

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 343 AKT1 (v-akt murine thymoma viral oncogene homolog 1) Etro D, et al.

al., 2003). AKT constitutive activation is observed in responsiveness to IGF-1 stimulation that leads to AKT more than 50% of AML cases and correlates with activation (Descamps et al., 2004). Furthermore, IL-6 chemotherapy resistance and poor prognosis (Min et and IGF-1 both upregulate telomerase activity, which is al., 2003; Grandage et al., 2005; Martelli et al., 2007). usually coupled with cell division, mediated by AKT In acute leukemias, AKT signaling (Akiyama et al., 2002). Constitutive activation gives rise to the upregulation of several phosphorylation of AKT has been reported in primary downstream targets as FoxO transcription factors in samples from patients with myeloma (Pene et al., AML patients with poor prognosis (Tamburini et al., 2002). In addition, inhibition of mTOR induces 2007; Cheong et al., 2003b), Bad, p27, GSK3beta, prevention in tumor proliferation and angiogenesis in IKK, p70S6K and 4E-BP1 in AML blasts (Zhao et al., myeloma cells associated with high levels of AKT 2004; Guzman et al., 2001; Xu et al., 2003). activation (Frost et al., 2004). AKT signaling plays an important role in cell survival Lymphomas mechanisms in acute promyelocytic leukemia (APL) Note (Billottet et al., 2009); recent advances have defined a AKT activation has been demonstrated in a variety of novel PML/PTEN/ AKT/mTOR/FoxO signaling B-cell non-Hodgkin's lymphomas (NHL) including network (Ito et al., 2009). The promyelocytic leukemia Follicular Lymphoma (FL), diffuse large B-cell protein (PML) has established activities as a potent lymphoma (DLBCL), marginal zone B-cell lymphoma repressor of proliferation and oncogenic and Mantle Cell Lymphoma (MCL) (Rudelius et al., transformation, a promoter of apoptosis, an inducer of 2006; Dal Col et al., 2008). Constitutive senescence, and may act as angiogenesis inhibitor. phosphorylation of AKT on serine 473 has been found PML tumour suppressor prevents cancer by also in peripheral leukemia cells of T-cell large inactivating phospho-AKT inside the nucleus and granular lymphocytic leukemia (T-LGL) (Schade et al., suppressing apoptotic rescue (Culjkovic et al., 2008). 2006). Constitutive phosphorylation of AKT, In acute lymphoblastic leukemia (ALL) cell lines such GSK3beta, and mTOR substrates such as S6K and 4E- as Jurkat T cells, PTEN is deleted thus activating the BP1 was demonstrated in Hodgkin's lymphoma (HL) AKT pathway and promoting survival (Xu et al., 2002; cell lines, suggesting that the AKT pathway plays a Uddin et al., 2004). In addition, an activating mutation crucial role in survival of HL cells (Dutton et al., of Notch1 receptor in ALL cells is found to inhibit 2005). Moreover, proteomic analysis of FL tissues PTEN expression with subsequent AKT activation showed overexpression of phospho-AKT on serine 473 (Palomero et al., 2007). (Gulman et al., 2005). In primary DLBCL samples, Chronic leukemia there is a correlation between poor prognosis and constitutive activation of AKT (Uddin et al., 2006; Note Ogasawara et al., 2003). In primary samples from Chronic myelogenous leukemia (CML) is caused by anaplastic large cell lymphoma (ALCL) patients, BCR-ABL fusion gene product, that has constitutive around half of ALCLs exhibit constitutive tyrosine kinase activity and evokes the PI3K/AKT phosphorylation of AKT on serine 473 and the AKT signaling pathway (Steelman et al., 2004). AKT is target p27 is downregulated in ALCL cell lines constitutively active in primary CML cells of both the (Rassidakis et al., 2005). Moreover, mTOR, S6K and chronic phase and blast crisis as well as in CML cell 4E-BP1 are constitutively phosphorylated in cell lines lines (Kawauchi et al., 2003). Introduction of a and in tissue samples from ALCL patients (Vega et al., dominant-negative kinase-deficient AKT mutant 2006), indicating that the AKT pathway may be (K179M) inhibits leukemo-genesis in murine cells, implicated in cell proliferation and survival of ALCL indicating an important role of AKT in transformation tumors. AKT and its downstream targets, including with BCR-ABL through the possible effectors FoxO, GSK3, FoxO3A, p27, MDM2, Bad, p70S6K and 4E- MDM2, GSK3beta, S6K and 4EBP-1 (Skorski et al, BP1, have been shown to be constitutively 1997; Kharas et al., 2005). Furthermore, AKT- phosphorylated in both primary MCL cells and MCL dependent phosphoryla-tion of FoxO3A is required for cell lines (Rudelius et al., 2006). AKT is likely to be maintaining the leukemic phenotype (Birkenkamp et more active in blastoid MCL variants than in typical al., 2007). MCL, suggesting that the AKT pathway plays a critical Myeloma role in pathogenesis in aggressive MCL cases. Constitutive AKT activation has been demonstrated in Note adult T-cell leukemic (ATL) cells as well as in ATL In both myeloma cell lines and primary cells IL-6 and cell lines. HSP90, a chaperone protein for AKT, and IGF-1 activate the PI3K/AKT pathway accompanied by the mTOR pathway are required for cell proliferation enhanced phosphorylation of downstream targets such and survival in primary ATL samples, suggesting a as Bad, GSK3beta, and FoxO (Hideshima et al., 2001; crucial role for the AKT/mTOR axis in ATL expansion Tu et al., 2000; Hsu et al., 2002). The expression of (Kawakami et al., 2007). B-cell antigen receptor (BCR) CD45 in myeloma cells negatively regulates the

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 344 AKT1 (v-akt murine thymoma viral oncogene homolog 1) Etro D, et al.

stimulation has been shown to induce AKT in AKT-mediated anti-apoptotic signaling (Kyoung phosphorylation on serine 473 (Poggi et al., 2008; Pyo et al., 2004). Longo et al., 2007). In addition, CpG- Schizophrenia oligodeoxynucleotide (CpG-ODN) stimulates leukemia cell proliferation accompanied by upregulation of AKT Note phosphorylation on 473 residue in B-CLL patients with Association between schizophrenia and an AKT1 poor prognosis (Longo et al., 2008). Therefore, AKT haplotype associated with lower AKT1 levels and a activation seems to be involved in CLL B-cell greater sensitivity to the sensorimotor gating-disruptive expansion. effect of amphetamine, conferred by AKT1 deficiency, Various diseases has been described. Alterations in AKT1/GSK3beta signaling contribute to schizophrenia pathogenesis and Note AKT1 gene may confer potential schizophrenia Alteration of AKT activity is associated with several susceptibility. Consistent with this proposal, it has been human diseases, including atherosclerosis, shown that haloperidol induces a stepwise increase in cardiovascular disease, Alzheimer disease, regulatory phosphorylation of AKT1 in the brains of schizophrenia and diabetes. treated mice, that could compensate for an impaired Atherosclerosis function of this signaling pathway in schizophrenia (Emamian et al., 2004). Note Oxidized low-density lipoproteins LDLs activate the Diabetes type 2 PI3K/AKT network in macrophages/foam cells (Biwa Note et al., 2000). The amount of phosphorylated AKT and AKT is involved in the pathomechanism of diabetes as other phosphorylated effector proteins as S6K, S6, it determines beta-cell apoptosis of Langerhans islets GSK3beta and FKHR was found to be reduced in and insulin sensitivity of the cells (Cseh et al., 2009; atherosclerotic lesions. Schulthess et al., 2009). It has been reported that Cardiovascular disease alterations of the AKT/mTOR or the AKT/PRAS40 axis contributes to a diabetic phenotype (Marshall et Note al., 2006; Nascimento et al., 2006). AKT is required for The first report on a role of the PI3K/AKT pathway in the metabolic actions of insulin; muscle cells from type the control of cell and organ size was published more 2 diabetic patients displayed defective insulin action than 10 years ago (Leevers et al., 1996). AKT signaling and a drastic reduction of insulin-stimulated activity of is relayed via mTOR to control the heart size. The all AKT isoforms, in particular with altered AKT1 cardiomyocyte-specific inactivation of the lipid phosphorylation on threonine 308 residue (Cozzone et phosphatase PTEN and subsequent AKT hyper- al., 2008). Insulin resistance can be induced by activation also triggers heart hypertrophy and stimulating the degradation of important molecules in culminates in reduced cardiac contractility (Crackower the insulin signaling pathway as AKT1 (Wing et al., et al., 2002). AKT is involved in the therapy for 2008). ischemic limb or heart (Huang et al., 2009; Kruger et al., 2009). Moreover, long-term activation of AKT/mTOR signaling links diet-induced obesity with References vascular senescence and cardiovascular disease (Wang Staal SP. Molecular cloning of the akt oncogene and its human et al., 2009). homologues AKT1 and AKT2: amplification of AKT1 in a primary human gastric adenocarcinoma. Proc Natl Acad Sci U Alzheimer disease S A. 1987 Jul;84(14):5034-7 Note Coffer PJ, Woodgett JR. Molecular cloning and Microtubule-associated protein tau contains a characterisation of a novel putative protein-serine kinase related to the cAMP-dependent and protein kinase C families. consensus motif for AKT encompassing the double Eur J Biochem. 1991 Oct 15;201(2):475-81 phospho-epitope (T212/S214). AKT dependent Knighton DR, Zheng JH, Ten Eyck LF, Ashford VA, Xuong NH, phosphorylation of tau occurs in vitro at both threonine Taylor SS, Sowadski JM. Crystal structure of the catalytic 212 and serine 214 and may play specific roles relevant subunit of cyclic adenosine monophosphate-dependent protein to Alzheimer disease and other neurodegenerations kinase. Science. 1991 Jul 26;253(5018):407-14 (Ksiezak-Reding et al., 2003). Modulators of the PI3K Bellacosa A, Franke TF, Gonzalez-Portal ME, Datta K, Taguchi pathway might be reduced during aging leading to a T, Gardner J, Cheng JQ, Testa JR, Tsichlis PN. Structure, sustained activation of GSK3beta, which in turn would expression and chromosomal mapping of c-akt: relationship to increase the risk of tau hyper-phosphorylation v-akt and its implications. Oncogene. 1993 Mar;8(3):745-54 (Mercado-Gomez et al., 2008). In primary cultures, Alessi DR, Andjelkovic M, Caudwell B, Cron P, Morrice N, AKT selectively phosphorylates tau at serine 214, Cohen P, Hemmings BA. Mechanism of activation of protein raising the possibility that 214 residue may participate kinase B by insulin and IGF-1. EMBO J. 1996 Dec 2;15(23):6541-51

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Diabetes. 2006 Jan 15;103(2):307-12 Dec;55(12):3221-8 Oki E, Baba H, Tokunaga E, Nakamura T, Ueda N, Futatsugi Rudelius M, Pittaluga S, Nishizuka S, Pham TH, Fend F, Jaffe M, Mashino K, Yamamoto M, Ikebe M, Kakeji Y, Maehara Y. ES, Quintanilla-Martinez L, Raffeld M. Constitutive activation of Akt phosphorylation associates with LOH of PTEN and leads to Akt contributes to the pathogenesis and survival of mantle cell chemoresistance for gastric cancer. Int J Cancer. 2005 Nov lymphoma. Blood. 2006 Sep 1;108(5):1668-76 10;117(3):376-80 Schade AE, Powers JJ, Wlodarski MW, Maciejewski JP. Palamarchuk A, Efanov A, Maximov V, Aqeilan RI, Croce CM, Phosphatidylinositol-3-phosphate kinase pathway activation Pekarsky Y. Akt phosphorylates and regulates Pdcd4 tumor protects leukemic large granular lymphocytes from undergoing suppressor protein. Cancer Res. 2005 Dec 15;65(24):11282-6 homeostatic apoptosis. 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Hepatocellular carcinoma: molecular Blood. 2006 Dec 15;108(13):4178-86 pathways and new therapeutic targets. Semin Liver Dis. 2005;25(2):212-25 Vega F, Medeiros LJ, Leventaki V, Atwell C, Cho-Vega JH, Tian L, Claret FX, Rassidakis GZ. Activation of mammalian Saji M, Vasko V, Kada F, Allbritton EH, Burman KD, Ringel target of rapamycin signaling pathway contributes to tumor cell MD. Akt1 contains a functional leucine-rich nuclear export survival in anaplastic lymphoma kinase-positive anaplastic sequence. Biochem Biophys Res Commun. 2005 Jun large cell lymphoma. Cancer Res. 2006 Jul 1;66(13):6589-97 24;332(1):167-73 Xuan Nguyen TL, Choi JW, Lee SB, Ye K, Woo SD, Lee KH, Sarbassov DD, Guertin DA, Ali SM, Sabatini DM. Ahn JY. Akt phosphorylation is essential for nuclear Phosphorylation and regulation of Akt/PKB by the rictor-mTOR translocation and retention in NGF-stimulated PC12 cells. complex. Science. 2005 Feb 18;307(5712):1098-101 Biochem Biophys Res Commun. 2006 Oct 20;349(2):789-98 Schwab SG, Hoefgen B, Hanses C, Hassenbach MB, Albus M, Birkenkamp KU, Essafi A, van der Vos KE, da Costa M, Hui Lerer B, Trixler M, Maier W, Wildenauer DB. Further evidence RC, Holstege F, Koenderman L, Lam EW, Coffer PJ. FOXO3a for association of variants in the AKT1 gene with schizophrenia induces differentiation of Bcr-Abl-transformed cells through in a sample of European sib-pair families. Biol Psychiatry. 2005 transcriptional down-regulation of Id1. J Biol Chem. 2007 Jan Sep 15;58(6):446-50 26;282(4):2211-20 Starkman BG, Cravero JD, Delcarlo M, Loeser RF. IGF-I Brognard J, Sierecki E, Gao T, Newton AC. PHLPP and a stimulation of proteoglycan synthesis by chondrocytes requires second isoform, PHLPP2, differentially attenuate the amplitude activation of the PI 3-kinase pathway but not ERK MAPK. of Akt signaling by regulating distinct Akt isoforms. Mol Cell. Biochem J. 2005 Aug 1;389(Pt 3):723-9 2007 Mar 23;25(6):917-31 Takeuchi H, Kim J, Fujimoto A, Umetani N, Mori T, Bilchik A, Carpten JD, Faber AL, Horn C, Donoho GP, Briggs SL, Turner R, Tran A, Kuo C, Hoon DS. X-Linked inhibitor of Robbins CM, Hostetter G, Boguslawski S, Moses TY, Savage apoptosis protein expression level in colorectal cancer is S, Uhlik M, Lin A, Du J, Qian YW, Zeckner DJ, Tucker-Kellogg regulated by hepatocyte growth factor/C-met pathway via Akt G, Touchman J, Patel K, Mousses S, Bittner M, Schevitz R, Lai signaling. Clin Cancer Res. 2005 Nov 1;11(21):7621-8 MH, Blanchard KL, Thomas JE. A transforming mutation in the Kornblau SM, Womble M, Qiu YH, Jackson CE, Chen W, pleckstrin homology domain of AKT1 in cancer. Nature. 2007 Konopleva M, Estey EH, Andreeff M. Simultaneous activation Jul 26;448(7152):439-44 of multiple signal transduction pathways confers poor Chan CB, Liu X, Tang X, Fu H, Ye K. Akt phosphorylation of prognosis in acute myelogenous leukemia. Blood. 2006 Oct zyxin mediates its interaction with acinus-S and prevents 1;108(7):2358-65 acinus-triggered chromatin condensation. Cell Death Differ. Li X, Lu Y, Jin W, Liang K, Mills GB, Fan Z. 2007 Sep;14(9):1688-99 Autophosphorylation of Akt at threonine 72 and serine 246. A Iantorno M, Chen H, Kim JA, Tesauro M, Lauro D, Cardillo C, potential mechanism of regulation of Akt kinase activity. J Biol Quon MJ. Ghrelin has novel vascular actions that mimic PI 3- Chem. 2006 May 12;281(19):13837-43 kinase-dependent actions of insulin to stimulate production of Marshall S. Role of insulin, adipocyte hormones, and nutrient- NO from endothelial cells. Am J Physiol Endocrinol Metab. sensing pathways in regulating fuel metabolism and energy 2007 Mar;292(3):E756-64 homeostasis: a nutritional perspective of diabetes, obesity, and Kawakami H, Tomita M, Okudaira T, Ishikawa C, Matsuda T, cancer. Sci STKE. 2006 Aug 1;2006(346):re7 Tanaka Y, Nakazato T, Taira N, Ohshiro K, Mori N. Inhibition of

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heat shock protein-90 modulates multiple functions required for Constitutively active Akt1 expression in mouse pancreas survival of human T-cell leukemia virus type I-infected T-cell requires S6 kinase 1 for insulinoma formation. J Clin Invest. lines and adult T-cell leukemia cells. Int J Cancer. 2007 Apr 2008 Nov;118(11):3629-38 15;120(8):1811-20 Autret A, Martin-Latil S, Brisac C, Mousson L, Colbère-Garapin Li X, Monks B, Ge Q, Birnbaum MJ. Akt/PKB regulates hepatic F, Blondel B. Early phosphatidylinositol 3-kinase/Akt pathway metabolism by directly inhibiting PGC-1alpha transcription activation limits poliovirus-induced JNK-mediated cell death. J coactivator. Nature. 2007 Jun 21;447(7147):1012-6 Virol. 2008 Apr;82(7):3796-802 Longo PG, Laurenti L, Gobessi S, Petlickovski A, Pelosi M, Bader AG, Vogt PK. Phosphorylation by Akt disables the anti- Chiusolo P, Sica S, Leone G, Efremov DG. The Akt signaling oncogenic activity of YB-1. Oncogene. 2008 Feb pathway determines the different proliferative capacity of 14;27(8):1179-82 chronic lymphocytic leukemia B-cells from patients with progressive and stable disease. Leukemia. 2007 Boormans JL, Hermans KG, van Leenders GJ, Trapman J, Jan;21(1):110-20 Verhagen PC. An activating mutation in AKT1 in human prostate cancer. Int J Cancer. 2008 Dec 1;123(11):2725-6 Manning BD, Cantley LC. AKT/PKB signaling: navigating downstream. Cell. 2007 Jun 29;129(7):1261-74 Chen KF, Yeh PY, Yeh KH, Lu YS, Huang SY, Cheng AL. Down-regulation of phospho-Akt is a major molecular Martelli AM, Tazzari PL, Evangelisti C, Chiarini F, Blalock WL, determinant of bortezomib-induced apoptosis in hepatocellular Billi AM, Manzoli L, McCubrey JA, Cocco L. Targeting the carcinoma cells. Cancer Res. 2008 Aug 15;68(16):6698-707 phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin module for acute myelogenous leukemia therapy: Cozzone D, Fröjdö S, Disse E, Debard C, Laville M, Pirola L, from bench to bedside. Curr Med Chem. 2007;14(19):2009-23 Vidal H. Isoform-specific defects of insulin stimulation of Akt/protein kinase B (PKB) in skeletal muscle cells from type 2 Mortenson MM, Galante JG, Gilad O, Schlieman MG, diabetic patients. Diabetologia. 2008 Mar;51(3):512-21 Virudachalam S, Kung HJ, Bold RJ. BCL-2 functions as an activator of the AKT signaling pathway in pancreatic cancer. J Culjkovic B, Tan K, Orolicki S, Amri A, Meloche S, Borden KL. Cell Biochem. 2007 Dec 1;102(5):1171-9 The eIF4E RNA regulon promotes the Akt signaling pathway. J Cell Biol. 2008 Apr 7;181(1):51-63 Ogino S, Kawasaki T, Ogawa A, Kirkner GJ, Loda M, Fuchs CS. 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(CAOV-3) viability, migration and proliferation in association Mutational loss of PTEN induces resistance to NOTCH1 with PI3-k/Akt activation. Cancer Invest. 2008 Oct;26(8):822-9 inhibition in T-cell leukemia. Nat Med. 2007 Oct;13(10):1203- Han EK, Mcgonigal T, Butler C, Giranda VL, Luo Y. 10 Characterization of Akt overexpression in MiaPaCa-2 cells: Tamburini J, Elie C, Bardet V, Chapuis N, Park S, Broët P, prohibitin is an Akt substrate both in vitro and in cells. Cornillet-Lefebvre P, Lioure B, Ugo V, Blanchet O, Ifrah N, Anticancer Res. 2008 Mar-Apr;28(2A):957-63 Witz F, Dreyfus F, Mayeux P, Lacombe C, Bouscary D. Kang ES, Han D, Park J, Kwak TK, Oh MA, Lee SA, Choi S, Constitutive phosphoinositide 3-kinase/Akt activation Park ZY, Kim Y, Lee JW. O-GlcNAc modulation at Akt1 Ser473 represents a favorable prognostic factor in de novo acute correlates with apoptosis of murine pancreatic beta cells. Exp myelogenous leukemia patients. Blood. 2007 Aug Cell Res. 2008 Jul 1;314(11-12):2238-48 1;110(3):1025-8 Kim EK, Yun SJ, Do KH, Kim MS, Cho M, Suh DS, Kim CD, Tang X, Jang SW, Wang X, Liu Z, Bahr SM, Sun SY, Brat D, Kim JH, Birnbaum MJ, Bae SS. Lysophosphatidic acid induces Gutmann DH, Ye K. Akt phosphorylation regulates the tumour- cell migration through the selective activation of Akt1. Exp Mol suppressor merlin through ubiquitination and degradation. Nat Med. 2008 Aug 31;40(4):445-52 Cell Biol. 2007 Oct;9(10):1199-207 Kim MS, Jeong EG, Yoo NJ, Lee SH. Mutational analysis of Thamilselvan V, Craig DH, Basson MD. FAK association with oncogenic AKT E17K mutation in common solid cancers and multiple signal proteins mediates pressure-induced colon acute leukaemias. Br J Cancer. 2008 May 6;98(9):1533-5 cancer cell adhesion via a Src-dependent PI3K/Akt pathway. FASEB J. 2007 Jun;21(8):1730-41 Liu Z, Hou P, Ji M, Guan H, Studeman K, Jensen K, Vasko V, El-Naggar AK, Xing M. Highly prevalent genetic alterations in Van Der Haar F. Goiter and other iodine deficiency disorders: a receptor tyrosine kinases and phosphatidylinositol 3-kinase/akt systematic review of epidemiological studies to deconstruct the and mitogen-activated protein kinase pathways in anaplastic complex web. Arch Med Res. 2007 Jul;38(5):586-7; author and follicular thyroid cancers. J Clin Endocrinol Metab. 2008 reply 588-9 Aug;93(8):3106-16 Varma S, Khandelwal RL. Effects of rapamycin on cell Lockwood WW, Chari R, Coe BP, Girard L, Macaulay C, Lam proliferation and phosphorylation of mTOR and p70(S6K) in S, Gazdar AF, Minna JD, Lam WL. DNA amplification is a HepG2 and HepG2 cells overexpressing constitutively active ubiquitous mechanism of oncogene activation in lung and other Akt/PKB. Biochim Biophys Acta. 2007 Jan;1770(1):71-8 cancers. Oncogene. 2008 Jul 31;27(33):4615-24 Zhu L, Hu C, Li J, Xue P, He X, Ge C, Qin W, Yao G, Gu J. Longo PG, Laurenti L, Gobessi S, Sica S, Leone G, Efremov Real-time imaging nuclear translocation of Akt1 in HCC cells. DG. The Akt/Mcl-1 pathway plays a prominent role in Biochem Biophys Res Commun. 2007 May 18;356(4):1038-43 mediating antiapoptotic signals downstream of the B-cell Alliouachene S, Tuttle RL, Boumard S, Lapointe T, Berissi S, receptor in chronic lymphocytic leukemia B cells. Blood. 2008 Germain S, Jaubert F, Tosh D, Birnbaum MJ, Pende M. Jan 15;111(2):846-55

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Ma S, Lee TK, Zheng BJ, Chan KW, Guan XY. CD133+ HCC Cseh A, Szebeni B, Szalay B, Vásárhelyi B. [Akt enzyme: new cancer stem cells confer chemoresistance by preferential therapeutic target in cancer and diabetes?]. Orv Hetil. 2009 expression of the Akt/PKB survival pathway. Oncogene. 2008 Feb 22;150(8):373-8 Mar 13;27(12):1749-58 Gallay N, Dos Santos C, Cuzin L, Bousquet M, Simmonet Malanga D, Scrima M, De Marco C, Fabiani F, De Rosa N, De Gouy V, Chaussade C, Attal M, Payrastre B, Demur C, Récher Gisi S, Malara N, Savino R, Rocco G, Chiappetta G, Franco R, C. The level of AKT phosphorylation on threonine 308 but not Tirino V, Pirozzi G, Viglietto G. Activating E17K mutation in the on serine 473 is associated with high-risk cytogenetics and gene encoding the protein kinase AKT1 in a subset of predicts poor overall survival in acute myeloid leukaemia. squamous cell carcinoma of the lung. Cell Cycle. 2008 Mar Leukemia. 2009 Jun;23(6):1029-38 1;7(5):665-9 Huang J, Inoue M, Hasegawa M, Tomihara K, Tanaka T, Chen Mercado-Gómez O, Hernández-Fonseca K, Villavicencio- J, Hamada H. Sendai viral vector mediated angiopoietin-1 Queijeiro A, Massieu L, Chimal-Monroy J, Arias C. Inhibition of gene transfer for experimental ischemic limb disease. Wnt and PI3K signaling modulates GSK-3beta activity and Angiogenesis. 2009;12(3):243-9 induces morphological changes in cortical neurons: role of tau phosphorylation. Neurochem Res. 2008 Aug;33(8):1599-609 Ito K, Bernardi R, Pandolfi PP. A novel signaling network as a critical rheostat for the biology and maintenance of the normal Okada M, Jang SW, Ye K. Akt phosphorylation and nuclear stem cell and the cancer-initiating cell. Curr Opin Genet Dev. phosphoinositide association mediate mRNA export and cell 2009 Feb;19(1):51-9 proliferation activities by ALY. Proc Natl Acad Sci U S A. 2008 Jun 24;105(25):8649-54 Krüger M, Linke WA.. Titin-based mechanical signalling in normal and failing myocardium. J Mol Cell Cardiol. 2009 Poggi A, Catellani S, Bruzzone A, Caligaris-Cappio F, Gobbi Apr;46(4):490-8. M, Zocchi MR. Lack of the leukocyte-associated Ig-like receptor-1 expression in high-risk chronic lymphocytic Qian J, Zou Y, Rahman JS, Lu B, Massion PP. Synergy leukaemia results in the absence of a negative signal between phosphatidylinositol 3-kinase/Akt pathway and Bcl-xL regulating kinase activation and cell division. Leukemia. 2008 in the control of apoptosis in adenocarcinoma cells of the lung. May;22(5):980-8 Mol Cancer Ther. 2009 Jan;8(1):101-9 Varma S, Khandelwal RL. Overexpression of Akt1 upregulates Schulthess FT, Paroni F, Sauter NS, Shu L, Ribaux P, Haataja glycogen synthase activity and phosphorylation of mTOR in L, Strieter RM, Oberholzer J, King CC, Maedler K. CXCL10 IRS-1 knockdown HepG2 cells. J Cell Biochem. 2008 Apr impairs beta cell function and viability in diabetes through 1;103(5):1424-37 TLR4 signaling. Cell Metab. 2009 Feb;9(2):125-39 Wing SS. The UPS in diabetes and obesity. BMC Biochem. Wang CY, Kim HH, Hiroi Y, Sawada N, Salomone S, Benjamin 2008 Oct 21;9 Suppl 1:S6 LE, Walsh K, Moskowitz MA, Liao JK. Obesity increases vascular senescence and susceptibility to ischemic injury Zhou QL, Jiang ZY, Holik J, Chawla A, Hagan GN, Leszyk through chronic activation of Akt and mTOR. Sci Signal. 2009 Mar 17;2(62):ra11 J, Czech MP. Akt substrate TBC1D1 regulates GLUT1 expression through the mTOR pathway in 3T3-L1 adipocytes. Yoeli-Lerner M, Chin YR, Hansen CK, Toker A. Akt/protein Biochem J. 2008 May 1;411(3):647-55 kinase b and glycogen synthase kinase-3beta signaling pathway regulates cell migration through the NFAT1 Billottet C, Banerjee L, Vanhaesebroeck B, Khwaja A. transcription factor. Mol Cancer Res. 2009 Mar;7(3):425-32 Inhibition of class I phosphoinositide 3-kinase activity impairs proliferation and triggers apoptosis in acute promyelocytic This article should be referenced as such: leukemia without affecting atra-induced differentiation. Cancer Res. 2009 Feb 1;69(3):1027-36 Etro D, Missiroli S, Buontempo F, Neri LM, Capitani S. AKT1 (v-akt murine thymoma viral oncogene homolog 1). Atlas Ch'ng S, Tan ST. Genetics, cellular biology and tumor Genet Cytogenet Oncol Haematol. 2010; 14(4):336-352. microenvironment of melanoma. Front Biosci. 2009 Jan 1;14:918-28

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Gene Section Mini Review

B3GNT6 (UDP-GlcNAc:betaGal beta-1,3-N- acetylglucosaminyltransferase 6 (core 3 synthase)) Neeru M Sharma, Prakash Radhakrishnan, Shuhua Tan, Pi-Wan Cheng Department of Biochemistry and Molecular Biology, College of Medicine, University of Nebraska Medical Center, 985870 Nebraska Medical Center, Omaha, NE 68198-5870, USA (NMS, PR, ST, PWC)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/B3GNT6ID44427ch11q13.html DOI: 10.4267/2042/44726 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Description Human B3GNT6 gene is 7,111 bp in length, composed Other names: B3Gn-T6; beta3GNT6; BGnT-6; Beta- of 2 exons and 1 intron, and located at chromosome 1,3-N-acetylglucosaminyltransferase-6; Beta3Gn-T6; 11q13.4. C3S; Core 3 synthase; MGC119334; MGC119336; MGC119337 Transcription HGNC (Hugo): B3GNT6 B3GNT6 transcript contains two exons. Exon 1 is 121 bp and exon 2 is 1,917 bp. The exon 2 contains 1,155 Location: 11q13.5 bp ORF and 762 bp 3'UTR. Local order: -NA- Pseudogene Note B3GNT6 is a single-pass type II membrane protein -NA- belonging to the glycosyltransferase 31 family. Protein DNA/RNA Note Note Human UDP-GlcNAc:GalNAca1-Ser/Thr beta-1,3-N- Human B3GNT6 is located on chromosome 11 in the acetylglucosaminyltransferase 6 (core 3 synthase) has region of q13.4. 384 amino acids and 43 KDa molecular weight.

The predicted B3GNT6 structure contains a short N-terminal cytoplasmic tail (CT) (12 aa), a transmembrane domain (TM) (19 aa), a long stem region and catalytic domain (353 aa) at the C-terminal region. Description

B3GNT6/C3S is single-pass type II membrane protein The schematic representation of human B3GNT6 gene and its transcript (ATG, translation start codon; TGA, translation stop belonging to the glycosyltransferase 31 family. codon; UTR, Untranslated region; ORF, Open reading frame).

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 353 B3GNT6 (UDP-GlcNAc:betaGal beta-1,3-N-acetylglucosaminyltransferase 6 (core 3 synthase)) Sharma NM, et al.

Expression mucus layer is one major cause of this cancer. Globally, cancer of the colon and rectum is the third B3GNT6/C3S gene expression is restricted to mucus- leading cause of cancer in males and the fourth leading secretory tissues. The level of beta3GnT6 transcript cause of cancer in females. The frequency of colorectal expressed in various human tissues as measured by the cancer varies around the world. It is common in the real time PCR revealed that the expression level was Western world and is rare in Asia and Africa. In highest in the stomach, followed by the colon and small countries where the people have adopted western diets, intestine. Skeletal muscle and testis expressed the the incidence of colorectal cancer is increasing. beta3GnT6 transcript at a moderate level. The Colorectal cancer can take many years to develop and expression levels in the remaining tissues were very early detection of colorectal cancer greatly improves low or not detectable. Its expression was markedly the chances of a cure. Therefore, screening for the down-regulated in gastric and colorectal carcinomas, disease is recommended in individuals who are at which include both tumor tissues and cell lines-derived increased risks. Prevention and early detection are key from these carcinomas. factors in controlling and curing colorectal cancer. Localisation Indeed, colorectal cancer is the second most Golgi Membrane. preventable cancer, after lung cancer. Function Prognosis B3GNT6/C3S is down-regulated in gastric and B3GNT6/C3S enzyme catalyzes the transfer of colorectal carcinomas, suggesting that it may be used GlcNAc from UDP-GlcNAc to GalNAcalpha1-Ser/Thr as a marker for distinguishing between benign (Tn antigen) to form the core 3 structure adenomas and premalignant lesions. (GlcNAcbeta1-3GalNAcalpha1-Ser/Thr). Core 3 can be extended by the addition of galactose and then other Oncogenesis sugars to generate biologically important epitopes or O-linked oligosaccharides (O-glycans) are the primary serves as the precursor for the formation of core 4, components of the intestinal mucus layer that covers which in turn can be further elaborated to form more the gastrointestinal epithelium. This layer is a dense, complex structure. Core 3-containing O-glycans are carbohydrate-rich matrix that consists primarily of found in the secreted mucins produced in the mucus- mucins containing multiple serine and threonine secretory tissues. Loss of core 3 synthase results in the residues, which have been modified by O-glycans and loss of not only core 3 glycans but also core 4 glycans. account for 80-90% of the mucin mass. The mucus Loss of core 3 could lead to the production of secreted layer and epithelial cells comprise an intestinal barrier mucins with compromised mucus protection function. that protects epithelial and intestinal mucosal immune As a result, mucus would be more dehydrated, bacteria cells from potentially harmful luminal microflora and would be inefficiently cleared from the system, and food components. Among all mucin glycan core chronic inflammation would be developed, which structures, Core 3 and core 4 are unique to secreted eventually would result in development of cancer. A mucins, which may play important roles in protecting mouse model devoid of core 3 synthase gene has been the molecular integrity of these mucins and enable shown to develop colon cancer. Because the loss of this them to perform their functions under extreme harsh gene leads to development of colon cancer, conditions, such as gastric and colonic environment. B3GNT6/C3S gene is a tumor suppressor gene. Loss of these functions resulted from the loss of core 3 synthase is thought to initiate oncogenesis in the Homology gastrointestinal tract. An alignment of the amino acid sequences of five B3GnTs made using ClustalW showed 41, 54, 42, and References 35% sequence identity between B3GnT6 and B3GnT2, Brockhausen I. Biosynthesis and functions of O-glycans and B3GnT3, B3GnT4, and B3GnT5, respectively, and this regulation of mucin antigen expression in cancer. Biochem Soc sequence similarity was limited to the putative catalytic Trans. 1997 Aug;25(3):871-4 domains. Five cysteine residues were conserved among Iwai T, Inaba N, Naundorf A, Zhang Y, Gotoh M, Iwasaki H, these five B3GnTs. However, only B3GNT6/C3S Kudo T, Togayachi A, Ishizuka Y, Nakanishi H, Narimatsu H. exhibits significant core 3 synthase activity. Molecular cloning and characterization of a novel UDP- GlcNAc:GalNAc-peptide beta1,3-N- acetylglucosaminyltransferase (beta 3Gn-T6), an enzyme Implicated in synthesizing the core 3 structure of O-glycans. J Biol Chem. Gastric and Colorectal carcinomas 2002 Apr 12;277(15):12802-9 Iwai T, Kudo T, Kawamoto R, Kubota T, Togayachi A, Hiruma Note T, Okada T, Kawamoto T, Morozumi K, Narimatsu H. Core 3 Colorectal cancer, which is also called colon cancer synthase is down-regulated in colon carcinoma and profoundly or large bowel cancer, includes cancerous growths in suppresses the metastatic potential of carcinoma cells. Proc the colon, rectum and appendix. Loss of function of the Natl Acad Sci U S A. 2005 Mar 22;102(12):4572-7

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 354 B3GNT6 (UDP-GlcNAc:betaGal beta-1,3-N-acetylglucosaminyltransferase 6 (core 3 synthase)) Sharma NM, et al.

Brockhausen I. Mucin-type O-glycans in human colon and This article should be referenced as such: breast cancer: glycodynamics and functions. EMBO Rep. 2006 Jun;7(6):599-604 Sharma NM, Radhakrishnan P, Tan S, Cheng PW. B3GNT6 (UDP-GlcNAc:betaGal beta-1,3-N- An G, Wei B, Xia B, McDaniel JM, Ju T, Cummings RD, Braun acetylglucosaminyltransferase 6 (core 3 synthase)). Atlas J, Xia L. Increased susceptibility to colitis and colorectal tumors Genet Cytogenet Oncol Haematol. 2010; 14(4):353-355. in mice lacking core 3-derived O-glycans. J Exp Med. 2007 Jun 11;204(6):1417-29

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Gene Section Review

BAX (BCL2-associated X protein) Hellinida Thomadaki, Andreas Scorilas Department of Biochemistry and Molecular Biology, University of Athens, 157 01, Panepistimiopolis, Athens, Greece (HT, AS)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/BAXID128ch19q13.html DOI: 10.4267/2042/44727 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Pseudogene Not identified so far. Other names: BCL2L4 HGNC (Hugo): BAX Protein Location: 19q13.33 Note Local order: Orientation: Plus Strand. The BAX gene encodes for a 21 kDa protein, named BAX-alpha. It was the first death-inducing member of DNA/RNA the BCL2 family to be identified, and it was detected as Description a protein co-purified with BCL2 in immunoprecipitation studies. The BH3 domain of BAX The BAX gene, with 6.939 bases in length, consists of is essential for its homodimerization and its 6 exons and 5 intervening introns. heterodimerization with BCL2 and BCL-XL. Transcription Furthermore, the protein contains a hydrophobic C- terminal region essential for membrane targeting, while The BAX gene is characterized by 5 protein coding BH1 and BH2 domains show homology to pore- transcripts (alpha/psi, beta, delta, epsilon, sigma). Bax- forming proteins that contribute to apoptosis. In beta encodes the longest isoform (891 bp) of the gene. addition to BAX-alpha, which is the major protein BAX-alpha/Bax-psi variant is 888 bp in length and product of the whole gene, BAX undergoes alternative codes for a protein isoform that possesses a shorter and splicing, resulting in the production of distinct protein different C terminus, as compared with the isoform isoforms. The tumour suppressor p53 is a BAX-beta. The third variant (BAX-delta), which is 741 transcriptional regulator of BAX, since the promoter of bp in length, lacks exon 3, whereas it retains the the BAX gene possesses four regions with high functionally critical C-terminal membrane anchorage homology to the consensus p53 binding sites. region, as well as the BCL2 homology 1 and 2 (BH1 and BH2) domains, although it has a shorter and Description different C terminus, in comparison with BAX-beta. The BAX belongs to the BCL2 family of proteins. It is The fourth identified variant of BAX, which is composed of 192 amino acids (21184 kDa), with a designated as BAX-epsilon, is 986 bp in length because calculated molecular mass of 21.184 kDa. The BAX it contains an extra fragment within the coding region, protein exists as a monomer, a homodimer, or as a as well as a distinct 3' coding region and 3' UTR, heterodimer with BCL2, E1B 19K protein, BCL2L1 resulting in a distinct BAX isoform with a shorter and isoform Bcl-X(L), MCL1 and BCL2A1/A1. It also distinct C terminus, as compared with BAX-beta. The interacts with SH3GLB1 and HN. It contains one BH3 fifth identified variant of BAX, BAX-sigma, is 849 bp homology domain. in length and has also a shorter and different C terminus, when compared with the isoform beta. Localisation BAX protein has been reported to be localized in the

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 356 BAX (BCL2-associated X protein) Thomadaki H, Scorilas A

mitochondria, mitochondrial permeability transition pore complex, mitochondrial outer membrane, Implicated in endoplasmic reticulum membrane and cytoplasm. Various cancers and diseases Function Disease BAX protein heterodimerizes either with members of Colorectal cancer, T-cell acute lymphoblastic leu- the BCL2 family of proteins or with tyrosine kinases kemia, chronic lymphocytic leukemia (CLL), B cell enabling them it to display its proapoptotic function chronic lymphocytic leukemia, osteomyelitis. within the cell. It is also implicated in the loss of Prognosis mitochondrial membrane potential and the release of BAX mutations have been found to be associated with cytochrome c. positive prognosis in Dukes B2 patients, concerning Homology survival. Human BAX shares 99.5% amino acid identity with The G(-248)A polymorphism in the promoter region of Pan troglodytes, 97.4% identity with Canis lupus the BAX gene has been associated with reduced BAX familiaris, 96.4% identity with Bos Taurus, 92.2% expression, advanced disease stage, reduced treatment identity with Mus musculus, 91.2% identity with Rattus response and short overall survival in B-cell chronic norvegicus and 52.7% identity with Danio rerio. In lymphocytic leukemia (CLL). addition, BAX protein presents high homology to the Polymorphisms were found for BAX, caused by BCL2 protein, containing the conserved regions BH1, variation in nucleotide A repeat number at position 360 BH2 and BH3. in the 5'-region of BAX gene. These allelic frequencies of BAX polymorphism were significantly different Mutations between males and females and therefore associated with gender-based heamatocrite (HCT) differences Note One regulatory type of mutation has been identified Substitution of the nucleotide G-->A at position -248 in according to which a guanine substituting adenosine the BAX gene was more frequent in patients with substitution at position 125 (G125A) in the BAX osteomyelitis and was associated with a longer lifespan promoter is associated with higher stage of chronic of their peripheral blood neutrophils, probably lymphocytic leukemia (CLL) and failure to respond to possessing a significant role in the pathogenesis of treatment in CLL patients. Additionally, 110 SNPs, osteomyelitis. with uknown clinical association and the following IDs, In cases of malignancies, the concentration of BAX have been reported in Entrez SNP database: protein in cancer cells is reduced. In addition, p53- rs62125987, rs62125961, rs61473366, rs61415800, deficient mice show reduced BAX levels, ultimately rs60900019, rs59878749, rs59152877, rs57453473, developing T-cell lymphoma. rs57028628, rs56251427, rs56251427, rs55692456, Reduction of BAX expression levels is negatively rs55692456, rs36101119, rs36096807, rs36017265, associated with many cancers outcome. It is associated rs35946201, rs35630245, rs35475300, rs35258702, with a variety of adverse prognostic factors such as rs34873472, rs34124134, rs34043541, rs28624947, poor response to radio- and chemotherapy, advanced rs28450536, rs12983717, rs12976339, rs12976283, stage, lymph node metastasis, and reduced disease-free rs12975003, rs11671610, rs11669164, rs11669162, and overall survival in variety cancer types, such as rs11668424, rs11668008, rs11667351, rs11400412, colorectal, pancreatic, breast, head and neck, prostate, rs11358529, rs11302449, rs10644606, rs7508566, small cell lung cancer and gynecological (ovarian) rs7259013, rs7255991, rs7255559, rs4645904, malignancies. More specifically, the enhanced rs4645903, rs4645902, rs4645903, rs4645902, expression of BAX protein is a positive prognostic rs4645901, rs4645900, rs4645899, rs4645898, factor for pancreatic cancer and sensitizes human rs4645897, rs4645896, rs4645895, rs4645894, pancreatic cancer cells to apoptosis induced by rs4645893, rs4645891, rs4645890, rs4645889, chemotherapeutic agents. In the case of stage II colon rs4645888, rs4645887, rs4645886, rs4645885, cancer, treated only with surgery, BAX protein rs4645884, rs4645883, rs4645882, rs4645881, expression may be a predictor for prognosis. In ovarian rs4645880, rs4645879, rs4645878, rs4309503, cancer, BAX protein may have a predictive potential in rs3817074, rs3817073, rs2387583, rs1985882, taxane-platinum-treated patients. Moreover, in resected rs1974820, rs1805419, rs1805418, rs1805417, non-small cell lung cancer, low expression of BAX rs1805416, rs1075531, rs1057369, rs1010104, implies poor prognosis. In addition, in patients with rs1010103, rs1009316, rs1009315, rs905238, rs704243. advanced esophageal cancer, treated with

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 357 BAX (BCL2-associated X protein) Thomadaki H, Scorilas A

chemoradiotherapy, reduced expression levels of BAX Rampino N, Yamamoto H, Ionov Y, Li Y, Sawai H, Reed JC, predict poor prognosis. Low expression of BAX was Perucho M. Somatic frameshift mutations in the BAX gene in colon cancers of the microsatellite mutator phenotype. also Science. 1997 Feb 14;275(5302):967-9 significantly associated with poor PFS and OS in Friess H, Lu Z, Graber HU, Zimmermann A, Adler G, Korc M, nasopharyngeal cancer patients. Schmid RM, Büchler MW. bax, but not bcl-2, influences the In lung cancer, BAX is translocated to the nucleus, prognosis of human pancreatic cancer. Gut. 1998 Sep;43(3):414-21 enhancing tumour development. Furthermore, mutational analysis of the gene in cases of lung cancer Meijerink JP, Mensink EJ, Wang K, Sedlak TW, Slöetjes AW, de Witte T, Waksman G, Korsmeyer SJ. Hematopoietic patients revealed the presence of a silent point mutation malignancies demonstrate loss-of-function mutations of BAX. in codon 184 (TCG>TCA), as well as intronic Blood. 1998 Apr 15;91(8):2991-7 mutations. Narita M, Shimizu S, Ito T, Chittenden T, Lutz RJ, Matsuda H, In T cells and endometrium of patients with acute Tsujimoto Y. Bax interacts with the permeability transition pore lymphoblastic leukaemia, frameshift mutations have to induce permeability transition and cytochrome c release in been detected in the BAX gene. isolated mitochondria. Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):14681-6 It is a common observation in cases of gastrointestinal Oliveira C, Seruca R, Seixas M, Sobrinho-Simões M. The cancer, the detection of two specific missense clinicopathological features of gastric carcinomas with mutations of the BAX gene in codon 169 (Thr > Ala or microsatellite instability may be mediated by mutations of Thr > Met), which cause inhibition of the proapoptotic different "target genes": a study of the TGFbeta RII, IGFII R, activity of the protein and enhance the development of and BAX genes. Am J Pathol. 1998 Oct;153(4):1211-9 cancer. Perez GI, Robles R, Knudson CM, Flaws JA, Korsmeyer SJ, Tilly JL. Prolongation of ovarian lifespan into advanced Various chemotherapeutic treatments act via up- chronological age by Bax-deficiency. Nat Genet. 1999 regulation of the BAX gene to block tumour Feb;21(2):200-3 progression. Shimizu S, Narita M, Tsujimoto Y. Bcl-2 family proteins BAX is highly expressed in HL-60 but it was found to regulate the release of apoptogenic cytochrome c by the be hardly expressed in HL-CR cells, a C2-ceramide- mitochondrial channel VDAC. Nature. 1999 Jun resistant HL-60 subline, which has been recently 3;399(6735):483-7 established. These cells showed reduced response to a Soini Y, Kinnula V, Kaarteenaho-Wiik R, Kurttila E, Linnainmaa variety of anticancer drugs including ceramide, K, Pääkkö P. Apoptosis and expression of apoptosis regulating proteins bcl-2, mcl-1, bcl-X, and bax in malignant doxorubicin, etoposide and cytosine arabinoside. mesothelioma. Clin Cancer Res. 1999 Nov;5(11):3508-15 Hybrid/Mutated gene Caligo MA, Ghimenti C, Sensi E, Marchetti A, Bertacca G, Not identified so far. Giulianotti PG, Fornaciari G, Bevilacqua G. Microsatellite Abnormal protein alterations and K-ras, TGFbetaRII, IGFRII and bax mutations in sporadic cancers of the gastrointestinal tract. Oncol Rep. Not identified so far. 2000 Nov-Dec;7(6):1371-5 Lindsten T, Ross AJ, King A, Zong WX, Rathmell JC, Shiels References HA, Ulrich E, Waymire KG, Mahar P, Frauwirth K, Chen Y, Wei M, Eng VM, Adelman DM, Simon MC, Ma A, Golden JA, Evan Korsmeyer SJ, Shutter JR, Veis DJ, Merry DE, Oltvai ZN. Bcl- G, Korsmeyer SJ, MacGregor GR, Thompson CB. The 2/Bax: a rheostat that regulates an anti-oxidant pathway and combined functions of proapoptotic Bcl-2 family members bak cell death. Semin Cancer Biol. 1993 Dec;4(6):327-32 and bax are essential for normal development of multiple Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes tissues. Mol Cell. 2000 Dec;6(6):1389-99 in vivo with a conserved homolog, Bax, that accelerates Suzuki M, Youle RJ, Tjandra N. Structure of Bax: coregulation programmed cell death. Cell. 1993 Aug 27;74(4):609-19 of dimer formation and intracellular localization. Cell. 2000 Nov Yin XM, Oltvai ZN, Korsmeyer SJ. BH1 and BH2 domains of 10;103(4):645-54 Bcl-2 are required for inhibition of apoptosis and Zhang L, Yu J, Park BH, Kinzler KW, Vogelstein B. Role of heterodimerization with Bax. Nature. 1994 May BAX in the apoptotic response to anticancer agents. Science. 26;369(6478):321-3 2000 Nov 3;290(5493):989-92 Apte SS, Mattei MG, Olsen BR. Mapping of the human BAX Antonsson B, Montessuit S, Sanchez B, Martinou JC. Bax is gene to chromosome 19q13.3-q13.4 and isolation of a novel present as a high molecular weight oligomer/complex in the alternatively spliced transcript, BAX delta. Genomics. 1995 Apr mitochondrial membrane of apoptotic cells. J Biol Chem. 2001 10;26(3):592-4 Apr 13;276(15):11615-23 Miyashita T, Reed JC. Tumor suppressor p53 is a direct Baxter RC. Signalling pathways involved in antiproliferative transcriptional activator of the human bax gene. Cell. 1995 Jan effects of IGFBP-3: a review. Mol Pathol. 2001 Jun;54(3):145-8 27;80(2):293-9 Matikainen T, Perez GI, Jurisicova A, Pru JK, Schlezinger JJ, Chou D, Miyashita T, Mohrenweiser HW, Ueki K, Kastury K, Ryu HY, Laine J, Sakai T, Korsmeyer SJ, Casper RF, Sherr Druck T, von Deimling A, Huebner K, Reed JC, Louis DN. The DH, Tilly JL. Aromatic hydrocarbon receptor-driven Bax gene BAX gene maps to the glioma candidate region at 19q13.3, but expression is required for premature ovarian failure caused by is not altered in human gliomas. Cancer Genet Cytogenet. biohazardous environmental chemicals. Nat Genet. 2001 1996 Jun;88(2):136-40 Aug;28(4):355-60

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Feb;157(1):18-24 Hum Mol Genet. 2002 Mar 15;11(6):675-87 Moshynska O, Moshynskyy I, Misra V, Saxena A. G125A Khaled AR, Li WQ, Huang J, Fry TJ, Khaled AS, Mackall CL, single-nucleotide polymorphism in the human BAX promoter Muegge K, Young HA, Durum SK. Bax deficiency partially affects gene expression. Oncogene. 2005 Mar 17;24(12):2042- corrects interleukin-7 receptor alpha deficiency. Immunity. 9 2002 Nov;17(5):561-73 Starczynski J, Pepper C, Pratt G, Hooper L, Thomas A, Kuhlmann T, Glas M, zum Bruch C, Mueller W, Weber A, Zipp Milligan D, Bentley P, Fegan C. Common polymorphism G(- F, Brück W. Investigation of bax, bcl-2, bcl-x and p53 gene 248)A in the promoter region of the bax gene results in polymorphisms in multiple sclerosis. J Neuroimmunol. 2002 significantly shorter survival in patients with chronic Aug;129(1-2):154-60 lymphocytic Leukemia once treatment is initiated. 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Ocaña MG, Valle-Garay E, Montes AH, Meana A, Cartón JA, Thomadaki H, Scorilas A. Molecular profile of breast versus Fierer J, Celada A, Asensi V. Bax gene G(-248)A promoter ovarian cancer cells in response to treatment with the polymorphism is associated with increased lifespan of the anticancer drugs cisplatin, carboplatin, doxorubicin, etoposide neutrophils of patients with osteomyelitis. Genet Med. 2007 and taxol. Biol Chem. 2008 Nov;389(11):1427-34 Apr;9(4):249-55 Fu NY, Sukumaran SK, Kerk SY, Yu VC. Baxbeta: a Thomadaki H, Scorilas A. Breast cancer cells response to the constitutively active human Bax isoform that is under tight antineoplastic agents cisplatin, carboplatin, and doxorubicin at regulatory control by the proteasomal degradation mechanism. the mRNA expression levels of distinct apoptosis-related Mol Cell. 2009 Jan 16;33(1):15-29 genes, including the new member, BCL2L12. Ann N Y Acad Sci. 2007 Jan;1095:35-44 Kim DH, Xu W, Ma C, Liu X, Siminovitch K, Messner HA, Lipton JH. Genetic variants in the candidate genes of the Willis SN, Fletcher JI, Kaufmann T, van Delft MF, Chen L, apoptosis pathway and susceptibility to chronic myeloid Czabotar PE, Ierino H, Lee EF, Fairlie WD, Bouillet P, Strasser leukemia. Blood. 2009 Mar 12;113(11):2517-25 A, Kluck RM, Adams JM, Huang DC. Apoptosis initiated when BH3 ligands engage multiple Bcl-2 homologs, not Bax or Bak. Li K, Lu Y, Liang J, Luo G, Ren G, Wang X, Fan D. RhoE Science. 2007 Feb 9;315(5813):856-9 enhances multidrug resistance of gastric cancer cells by suppressing Bax. Biochem Biophys Res Commun. 2009 Feb Wincewicz A, Sulkowska M, Koda M, Kanczuga-Koda L, 6;379(2):212-6 Witkowska E, Sulkowski S. Significant coexpression of GLUT- 1, Bcl-xL, and Bax in colorectal cancer. Ann N Y Acad Sci. Nehls O, Hass HG, Okech T, Zenner S, Hsieh CJ, Sarbia M, 2007 Jan;1095:53-61 Borchard F, Gruenagel HH, Gaco V, Porschen R, Gregor M, Klump B. Prognostic implications of BAX protein expression Garrofé-Ochoa X, Melero-Fernández de Mera RM, Fernández- and microsatellite instability in all non-metastatic stages of Gómez FJ, Ribas J, Jordán J, Boix J. BAX and BAK proteins primary colon cancer treated by surgery alone. Int J Colorectal are required for cyclin-dependent kinase inhibitory drugs to Dis. 2009 Jun;24(6):655-63 cause apoptosis. Mol Cancer Ther. 2008 Dec;7(12):3800-6 Schlauder SM, Calder KB, Khalil FK, Passmore L, Mathew RA, Gavathiotis E, Suzuki M, Davis ML, Pitter K, Bird GH, Katz SG, Morgan MB. Bif-1 and Bax expression in cutaneous Merkel cell Tu HC, Kim H, Cheng EH, Tjandra N, Walensky LD. BAX carcinoma. J Cutan Pathol. 2009 Jan;36(1):21-5 activation is initiated at a novel interaction site. Nature. 2008 Oct 23;455(7216):1076-81 Ziółkowska-Seta I, Madry R, Kraszewska E, Szyma ńska T, Timorek A, Rembiszewska A, Kupryja ńczyk J. TP53, BCL-2 Jeong SH, Lee HW, Han JH, Kang SY, Choi JH, Jung YM, and BAX analysis in 199 ovarian cancer patients treated with Choi H, Oh YT, Park KJ, Hwang SC, Sheen SS, Oh YJ, Kim taxane-platinum regimens. Gynecol Oncol. 2009 JH, Lim HY. Low expression of Bax predicts poor prognosis in Jan;112(1):179-84 resected non-small cell lung cancer patients with non- squamous histology. Jpn J Clin Oncol. 2008 Oct;38(10):661-9 This article should be referenced as such: Thomadaki H, Scorilas A. BAX (BCL2-associated X protein). Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4):356-360.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 360 Atlas of Genetics and Cytogenetics

in Oncology and Haematology OPEN ACCESS JOURNAL AT INIST-CNRS

Gene Section Review

CEACAM1 (carcinoembryonic antigen-related cell adhesion molecule 1 (biliary glycoprotein)) Yasunobu Matsuda Department of Medical Technology, Niigata University Graduate School of Health Sciences, Asahimachi- dori 2-746, Niigata 9518518, Japan (YM)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/CEACAM1ID40044ch19q13.html DOI: 10.4267/2042/44728 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Transcription Human CEACAM1 consists of 9 exons that can be Other names: BGP; BGPI; BGP1; CD66a; BGP alternatively spliced to generate 11 different isoforms. HGNC (Hugo): CEACAM1 The gene contains 19 different introns, and the Location: 19q13.2 sequence is supported by 286 sequences from 262 cDNA clones. The transcripts differ by truncation of Local order: centromere - CGM7(CEACAM4) - the N-terminus, truncation of the C-terminus, presence CGM2(CEACAM7) - CEA(CEACAM5) - or absence of a cassette exon, common exons with NCA(CEACAM6) - CGM1(CEACAM3) - CEACAM1 different boundaries. Among various types of isoforms, - CGM9(CEACAMP2) - CGM6(CEACAM8) - the full-length nature of two alternative transcripts CGM8(CEACAMP1) - telomere. (NM_001024912.1, NM_001712.3) has been DNA/RNA determined. Pseudogene Note Eleven pseudogenes of the CEA cell adhesion molecule The locus is complex, and it produces several proteins subgroup are found in the cluster of the two subgroups via alternative splicing. of the CEA family (the CEA cell adhesion molecules Description and the pregnancy-specific glycoproteins which locate The gene is composed of 9 exons spanning in a region within a 1.2 Mb cluster on the long arm of chromosome of 23,140 bp (Localized from 47,724,479 to 47,703,298 19). from pter).

Genomic structure of human CEACAM1. Open and closed boxes indicate untranslated regions and coding regions (exons) of CEACAM1 gene, respectively. The ATG initiation codon is located in the first exon, and the TGA termination codon is located in exon 9. N, A1, B1 and A2; Ig domains, TM; transmembrane domain, C; cytoplasmic domain.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 361 CEACAM1 (carcinoembryonic antigen-related cell adhesion molecule 1 (biliary glycoprotein)) Matsuda Y

In human, 11 different CEACAM1 splice variants have been reported. The domain structure of each protein isoform is illustrated as open box. N; N-terminal immunoglobulin variable-region-like (IgV-like) domain, A or B; subsets of constant-region-type-2-like (IgC2-like) domains, TM; transmembrane domain, C; cytoplasmic domain. Nomenclature number after CEACAM1 indicates the number of extracellular immunoglobulin-like domains, whereas the letter that follows this indicates the presence of either a long (L) or a short (S) cytoplasmic tail, a unique termini (C), or an Alu family repeat sequence present within the open reading frame (A). For example, CEACAM1 with four extracellular immunoglobulin-like domains and with a long cytoplasmic domain is called as CEACAM1-4L. The most common isoforms expressed by human immune cells are CEACAM1-4L, CEACAM1-3L, CEACAM1-4S and CEACAM1-3S.

has been implicated in various types of intercellular- Protein adhesion and intracellular-signaling of cell survival, Description differentia-tion and growth in both normal and cancer cells as follows: CEACAM1 is a single-pass type-1 glycoprotein which belongs within the Ig gene superfamily. It contains a 34 (1) CEACAM1 inhibits proliferation of both epithelial amino acid (aa) leader sequence, an extracellular cells and T lymphocytes by a contact-mediated domain which contains a N-terminal IgV-like domain mechanism. In monocytes, CEACAM1 regulates and three of IgC2-like domains (382 aa), a phosphatidylinositol 3-kinase (PI3K) and Akt- transmembrane domain (43 aa), a cytoplasmic domain dependent survival signal and inhibits apoptosis. (67 aa) and two of the potential tyrosine (2) CEACAM1 induces apoptosis of mature phosphorylation sites. CEACAM1 is heavily N- colonocytes and mammary epithelial cells, which is in glycosylated with more than 60% of the mass part of a morphogenetic process in the formation of contributed by N-linked glycans. glandular lumen. Localisation (3) CEACAM1 has been demonstrated to stimulate cellular migration of blood vessel endothelial cells. CEACAM1 isoforms with transmembrane domain are mostly cell-membrane anchored, whereas those without (4) In granulocytes, CEACAM1 mediates the binding transmembrane domain are secreted. They are mostly to endothelial E-selectin. expressed in cells of epithelial and endothelial, (5) The involvement of CEACAM1 in carcinogenesis lymphoid and myeloid cells. is complex. In some types of cancer cells such as colon Function cancer and hepatoma loss or reduced expression of CEACAM1 is frequently observed, thereby it has been CEACAM1 is capable of homophilic with CEACAM1 regarded as a tumor suppressor. However, in other or heterophilic adhesion with CEA and CEACAM6. types, for example, thyroid cancer and gastric cancer, Isoforms of CEACAM1 with a long cytoplasmic CEACAM1 is up-regulated. domain generally transmit inhibitory signals, whereas (6) CEACAM1 exerts angiogenic properties and is a isoforms with a short cytoplasmic domain indicate a major effecter of VEGF in the early vascular formation. regulated interaction with the cytoskeleton. CEACAM1

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 362 CEACAM1 (carcinoembryonic antigen-related cell adhesion molecule 1 (biliary glycoprotein)) Matsuda Y

Since CEACAM1 is mainly expressed in tumor Breast cancer microvessels but not in large blood vessels, it may be a Note target for the therapy of tumor angio-genesis. CEACAM1 is down-regulated in around 30% of breast (7) Neisseria gonorrhea triggers expression of cancers. CEACAM1 is reported to suppress the CEACAM1 on primary endothelial cells by activating tumorigenicity of breast cancer cells via its cytoplasmic the immediate early response trans-cription factor NF- domain but not the extracellular domain (Luo et al., kappaB. 1997). CEACAM1 is demonstrated to be consistently Neisseria gonorrhoeae colony opacity-associated (Opa) expressed in normal tissue and benign lesions of proteins bind to human CEACAM on host cells mammary gland, whereas it disappears with the including T lymphocytes, and Opa binding to development of the malignant phenotype in both CEACAM1 suppresses the activation of CD4+ T cells noninvasive and invasive carcinoma (Riethdorf et al., in response to a variety of stimuli.(8) CEACAM1 1997). CEACAM1 is reported to be expressed at high regulates insulin clearance in the liver. levels in cribiform ductal carcinoma in situ (DCIS). Homology Although most invasive ductal carcinomas express CEACAM1 weakly, tumours with minimal lumena CEACAM1 is the human homolog of a cell adhesion formation show cytoplasmic CEACAM1 expression molecule (CAM) of the rat designated Cell-CAM. (Kirshner et al., 2004). Rattus norvegicus: 70.74 (n), 56.12(a) (Percentage Prostate cancer Identity). Note Mus musculus: 72.3 (n), 58.25 (a). CEACAM1 is reported to be regulated by androgen and Implicated in might act as a growth repressor during differentiation of the prostatic epithelium. Forced expression of Colon cancer CEACAM1 results in the significan-tly lower growth rates of human prostatic cancer cell line PC-3 (Hsieh et Note al., 1995). CEACAM1 mRNA was demonstrated to be down- regulated to < or = 0.4 in 80% of 21 colorectal Bladder cancer carcinoma tissue specimens as compared with the Note respective adjacent normal mucosa (Neumaier et al., CEACAM1 is expressed in umbrella cells of bladder 1993). CEACAM1 was reported to be associa-ted with urothelium, but is down-regulated in superficial bladder pp60c-src, suggesting that down-regulation of cancer. CEACAM1 silencing in bladder cancer cell CEACAM1 in about 80% of colorectal carcino-mas lines 486p and RT4 using the small interfering RNA may contribute to a dysfunction of pp60c-src in technique leads to a significant up-regulation of colorectal cancer (Brummer et al., 1995). vascular endothelial growth factor (VEGF)-C and Hepatocellular carcinoma VEGF-D, suggesting that the epithelial down- regulation of CEACAM1 induces angiogenesis via Note increased expression of VEGF-C and VEGF-D CEACAM1 was reported to be diffusely expressed in (Oliveira-Ferrer et al., 2004). 113 of 139 hepatomas. Loss of CEACAM1 expression was closely associated with large tumor size, Melanoma multiplicity of the tumor, portal vein invasion, and Note satellite nodules and affected survival adversely, A total of 28 of 40 patients with CEACAM1-positive according to univariate (P < 0.0001) and multivariate primary melanomas developed metastatic disease, analyses (relative risk, 5.737; P < 0.001) (Cruz et al., compared with only six of 60 patients with 2005). In rat hepatoma cells 1682A, restoration of CEACAM1-negative melanomas. the strongest CEACAM1-4L expression was demonstrated to CEACAM1 expression was observed at the invading completely suppress the tumor formation (Laurie et al., front. Kaplan-Meier analysis revealed a highly 2005). In human hepatoma cell lines (HLF, significant association between CEACAM1 expression PLC/PRF/5, HepG2 and KYN-2), CEACAM1 protein and metastasis (P <.0001) (Thies et al., 2002). was demonstrated to only express in HepG2, which shows a strong property for enhanced anchorage- Renal cell carcinoma independent growth. It was reported that CEACAM1 Note acts as a tumor suppressor when the cells are cultured In normal kidney, CEACAM1 was found in epithelial in anchorage-dependent growth conditions. In contrast, cells of proximal tubules and in endothel-ial cells. In in anchorage-independent growth conditions, CEACAM1 augments cell proliferation by potentiating contrast, tumour cells of 30 clear cell, three the cell-cell attachment (Hokari et al., 2007). chromophobic, and two chromophilic RCCs were

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 363 CEACAM1 (carcinoembryonic antigen-related cell adhesion molecule 1 (biliary glycoprotein)) Matsuda Y

completely devoid of CEACAM1. Renal adenomas Wagener C, Ergün S. Angiogenic properties of the also lacked CEACAM1 expression. Similarly, RCC carcinoembryonic antigen-related cell adhesion molecule 1. Exp Cell Res. 2000 Nov 25;261(1):19-24 cell lines CaKi1, CaKi2, A498, and RCC26 exhibited no or low-level CEACAM1 expression. However, Poy MN, Yang Y, Rezaei K, Fernström MA, Lee AD, Kido Y, Erickson SK, Najjar SM. CEACAM1 regulates insulin clearance CEACAM1 expression was transiently induced in in liver. Nat Genet. 2002 Mar;30(3):270-6 A498 cells upon contact with allogeneic CD8+ T cells, mediated at least in part by interferon-gamma. Thies A, Moll I, Berger J, Wagener C, Brümmer J, Schulze HJ, Brunner G, Schumacher U. CEACAM1 expression in Furthermore, the majority of tumour-infiltrating T and cutaneous malignant melanoma predicts the development of NK cells expressed CEACAM1 upon stimulation. metastatic disease. J Clin Oncol. 2002 May 15;20(10):2530-6 Thus, transient expression of the tumour suppressor Abou-Rjaily GA, Lee SJ, May D, Al-Share QY, Deangelis AM, CEACAM1 by tumour cells and subsequent Ruch RJ, Neumaier M, Kalthoff H, Lin SH, Najjar SM. homophilic interaction with CEACAM1 on tumour- CEACAM1 modulates epidermal growth factor receptor-- infiltrating lympho-cytes could represent a novel mediated cell proliferation. J Clin Invest. 2004 Oct;114(7):944- immune escape mechanism in RCC (Kammerer et al., 52 2004). Kammerer R, Riesenberg R, Weiler C, Lohrmann J, Schleypen J, Zimmermann W. The tumour suppressor gene CEACAM1 is completely but reversibly downregulated in renal cell To be noted carcinoma. J Pathol. 2004 Nov;204(3):258-67 Note Kirshner J, Hardy J, Wilczynski S, Shively JE. Cell-cell The role of CEACAM1 in cancer may be different adhesion molecule CEACAM1 is expressed in normal breast and milk and associates with beta1 integrin in a 3D model of among different types of cancer. CEACAM1 is down- morphogenesis. J Mol Histol. 2004 Mar;35(3):287-99 regulated in colon carcinomas, hepatocellular carcinomas, a proportion of breast cancers. In contrast, Oliveira-Ferrer L, Tilki D, Ziegeler G, Hauschild J, Loges S, Irmak S, Kilic E, Huland H, Friedrich M, Ergün S. Dual role of in prostate cancer and bladder cancer, CEACAM1 is carcinoembryonic antigen-related cell adhesion molecule 1 in down-regulated in the cancer cells, whereas angiogenesis and invasion of human urinary bladder cancer. overexpressed in blood microvessels and lymphatic Cancer Res. 2004 Dec 15;64(24):8932-8 capillaries and in the vicinity of the tumor. In thyroid Cruz PV, Wakai T, Shirai Y, Yokoyama N, Hatakeyama K. cancer, gastric cancer and metastasing malignant Loss of carcinoembryonic antigen-related cell adhesion melanomas, CEACAM1 is increased. molecule 1 expression is an adverse prognostic factor in hepatocellular carcinoma. Cancer. 2005 Jul 15;104(2):354-60 References Laurie NA, Comegys MM, Carreiro MP, Brown JF, Flanagan DL, Brilliant KE, Hixson DC. Carcinoembryonic antigen-related Koelma IA, Nap M, Huitema S, Krom RA, Houthoff HJ. cell adhesion molecule 1a-4L suppression of rat hepatocellular Hepatocellular carcinoma, adenoma, and focal nodular carcinomas. Cancer Res. 2005 Dec 1;65(23):11010-7 hyperplasia. Comparative histopathologic study with immunohistochemical parameters. Arch Pathol Lab Med. 1986 Gray-Owen SD, Blumberg RS. CEACAM1: contact-dependent Nov;110(11):1035-40 control of immunity. Nat Rev Immunol. 2006 Jun;6(6):433-46 Neumaier M, Paululat S, Chan A, Matthaes P, Wagener C. Horst AK, Ito WD, Dabelstein J, Schumacher U, Sander H, Biliary glycoprotein, a potential human cell adhesion molecule, Turbide C, Brümmer J, Meinertz T, Beauchemin N, Wagener is down-regulated in colorectal carcinomas. Proc Natl Acad Sci C. Carcinoembryonic antigen-related cell adhesion molecule 1 U S A. 1993 Nov 15;90(22):10744-8 modulates vascular remodeling in vitro and in vivo. J Clin Invest. 2006 Jun;116(6):1596-605 Brümmer J, Neumaier M, Göpfert C, Wagener C. Association of pp60c-src with biliary glycoprotein (CD66a), an adhesion Kuespert K, Pils S, Hauck CR. CEACAMs: their role in molecule of the carcinoembryonic antigen family physiology and pathophysiology. Curr Opin Cell Biol. 2006 downregulated in colorectal carcinomas. Oncogene. 1995 Oct Oct;18(5):565-71 19;11(8):1649-55 Hokari M, Matsuda Y, Wakai T, Shirai Y, Sato M, Tsuchiya A, Hsieh JT, Luo W, Song W, Wang Y, Kleinerman DI, Van NT, Takamura M, Yamagiwa S, Suzuki K, Ohkoshi S, Ichida T, Lin SH. Tumor suppressive role of an androgen-regulated Kawachi H, Aoyagi Y. Tumor suppressor carcinoembryonic epithelial cell adhesion molecule (C-CAM) in prostate antigen-related cell adhesion molecule 1 potentates the carcinoma cell revealed by sense and antisense approaches. anchorage-independent growth of human hepatoma HepG2 Cancer Res. 1995 Jan 1;55(1):190-7 cells. Life Sci. 2007 Jul 4;81(4):336-45 Luo W, Wood CG, Earley K, Hung MC, Lin SH. Suppression of Slevogt H, Zabel S, Opitz B, Hocke A, Eitel J, N'guessan PD, tumorigenicity of breast cancer cells by an epithelial cell Lucka L, Riesbeck K, Zimmermann W, Zweigner J, adhesion molecule (C-CAM1): the adhesion and growth Temmesfeld-Wollbrueck B, Suttorp N, Singer BB. CEACAM1 suppression are mediated by different domains. Oncogene. inhibits Toll-like receptor 2-triggered antibacterial responses of 1997 Apr 10;14(14):1697-704 human pulmonary epithelial cells. Nat Immunol. 2008 Nov;9(11):1270-8 Riethdorf L, Lisboa BW, Henkel U, Naumann M, Wagener C, Löning T. Differential expression of CD66a (BGP), a cell This article should be referenced as such: adhesion molecule of the carcinoembryonic antigen family, in benign, premalignant, and malignant lesions of the human Matsuda Y. CEACAM1 (carcinoembryonic antigen-related cell mammary gland. J Histochem Cytochem. 1997 Jul;45(7):957- adhesion molecule 1 (biliary glycoprotein)). Atlas Genet 63 Cytogenet Oncol Haematol. 2010; 14(4):361-364.

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Gene Section Review

DAB2 (disabled homolog 2, mitogen-responsive phosphoprotein (Drosophila)) Maurizio Orlandini Department of Molecular Biology, Via Fiorentina 1, 53100 - Siena, Italy (MO)

Published in Atlas Database: May 2009 Online updated version : http://AtlasGeneticsOncology.org/Genes/DAB2ID40258ch5p13.html DOI: 10.4267/2042/44729 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Schematic representation of DAB2 domains. DAB2 possess a Other names: DAB-2; DOC-2; DOC2; FLJ26626 highly conserved N-terminal phosphotyrosine- HGNC (Hugo): DAB2 interacting/phosphotyrosine binding domain (PID/PTB), renamed the DAB homology domain, and a C-terminal proline- Location: 5p13.1 rich domain (PRD). Local order: The complement factor 9 (C9) gene is Description located at the 3'-end of the DAB2 gene. 770 amino acids, molecular weight 82.5 kDa. DAB2 Note contains an N-terminal PID/PTB domain (amino acid DAB2 was first identified as DOC-2, for differentially 42-180) and three C-terminal proline-rich domains expressed in ovarian carcinoma, and subsequently as a (amino acid 619-627, 663-671, and 714-722). A protein whose phosphorylation is stimulated by CSF-1. potential actin-binding motif, KKEK is present in the DNA/RNA N-terminal domain. Expression Description Widely expressed. Cytoplasmic. DAB2 is expres-sed in The DAB2 gene consists of 15 exons and 14 introns many epithelial cell types and was suggested to have a spanning in a region of 35 kb in size. role in epithelial organization. dab2 knock-out mice are Transcription embryonic lethal for defective visceral endoderm cell organization. In fact, in dab2 (-/-) mice, the epithelial The putative DAB2 promoter was identified within a cells of the early embryos (visceral endoderm) mix 420-bp sequence upstream of the exon1/intron1 within the interior rather then align as a layer covering junction. DAB2 is alternatively spliced to generate the inner cell mass. The role of Dab2 in mediating several transcripts and proteins. The transcript has been directional trafficking of endocytic proteins to establish detected in spleen, thymus, prostate, testis, small apical polarity is suggested as a mechanism for surface intestine, and abundant in ovary. positioning of endoderm cells. Protein Function Note The PID/PTB and PRD domains of DAB2 associate DAB2 plays a pivotal role in the control of cellular with several proteins, and these interactions have been homeostasis. The adaptor protein DAB2 is implicated shown to modulate protein trafficking, in several receptor-mediated signaling pathways, cytoskeleton organization, cell adhesion and migra- endocytosis, cell adhesive function, hematopoietic cell tion, and cell signaling of various receptor protein- differentiation, and angiogenesis. tyrosine kinases.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 365 DAB2 (disabled homolog 2, mitogen-responsive phosphoprotein (Drosophila)) Orlandini M

Cell cycle : DAB2 was identified as a protein integrin. In these conditions, Dab2 silencing leads to a phosphorylated in response to mitogenic stimula-tion decrease in cell adherence, inhibition of EMT, and by CSF-1. In cells, protein phosphorylation of DAB2 apoptosis. modulates its functional activity. Protein kinase C Angiogenesis : DAB2 can bind to Shc3 domain of Src (PKC) and Cdc2 are the two known DAB2 kinases. and this interaction results in Src inactivation. DAB2 is The major PKC phosphorylation site has been mapped 24 expressed in human umbilical vein endothelial cells to Ser and it is essential for the inhibitory function of (HUVEC). By modulating the activation of Src-FAK DAB2 in TPA-induced AP-1 gene transcription. DAB2 signaling and MAPK phosphorylation, DAB2 controls is differentially phosphorylated during the cell cycle by endothelial cell migration and differentiation. cdc2, and its phosphorylation promotes the association of DAB2 with Pin1, that regulates the rate of DAB2 Homology dephosphorylation. The Disabled proteins are a family of adapters involved Vesicle traffic : DAB2 plays a role in linking specific in cellular signaling, oncogenesis, and development. extracellular receptors to the endocytic machinery. DAB2 is related to Drosophila Disabled and DAB2 associates with AP-2-positive clathrin-coated mammalian Dab1, which regulate neuronal structures, together with endocy-tosed trans-membrane development. DAB2 shares 81% identity with the proteins such as low-density lipoprotein (LDL) mouse p96/Dab2 protein. receptors and integrins. DAB2 also binds to the actin- based myosin VI, mediating the attachment of cargos to Implicated in motor proteins and regulating protein trafficking. Epithelial ovarian cancer Signaling pathways: Note - TGFbeta - Dab2 associates with Smad2 and Smad3, DAB2 was identified due to the loss of its expression in by a direct interaction with the PID/PTB domain of ovarian cancer cells. Ovarian carcinoma cells Dab2, and with TGFbeta receptor I and TGFbeta transfected with DAB2 showed a reduced growth rate receptor II. Thus Dab2 may be an essential component and ability to form tumors in nude mice. Loss of DAB2 of the TGFbeta signaling pathway allowing the expression is not correlated with tumor grade, transmission of signals from the TGFbeta receptors to suggesting that loss of DAB2 expression is an early the Smad family of transcriptional activators. event in ovarian malignancies and DAB2 behaves as a - WNT - Dab2 associates with Axin and stabilizes its tumor suppressor. expression by preventing Axin interaction with the LRP5 co-receptor. Thus the interaction of Axin with Prostate cancer beta-catenin results stabilized with an increase in beta- Note catenin degradation and attenuation of Wnt signaling. DAB2 is a potent growth inhibitor for prostate cancer - RAS/RAF/MAPK - In cell culture experiments, a cells by suppressing several protein kinase pathways. Dab2 over-expression leads to suppression of c-Fos The PRD of DAB2 is the key functional domain expression and cell growth inhibition without affecting responsible for this activity. It was shown that in MAPK activity. In vivo studies confirmed a Dab2 role prostate cancer cells without endogenous DAB2 in regulating c-Fos expression. A possible molecular expression, a functional motif derived from the PRD of mechanism of action is that Dab2 limits the entry of the DAB2 conjugated with a delivery system is a potent activated MAPK into the nucleus. DAB2 can also growth inhibitor. interact with Grb2 through its PRD. Receptor tyrosine Breast cancer kinase activation by growth factors increases the binding of DAB2 to Grb2, which interrupts the binding Note of SOS to Grb2 and leads to suppression of ERK DAB2 sensitizes breast cancer cells to cell death upon activation. the loss of cell-matrix attachment by targeting the oncogenic activity of ILK. Cell adhesion : DAB2 is an adhesion-responsive phosphoprotein and plays a role in cell adhesion and Various cancer spreading. Ser 24 phosphorylation promotes membrane Note translocation of DAB2 and its interaction with beta3 Urothelial carcinoma of the bladder, esophageal integrin. DAB2 negatively regulates integrin squamous carcinoma, metastatic pancreatic cancer, alphaIIbbeta3 activation, leading to the inhibition of colorectal cancer, gestational choriocarcinoma. alphaIIbbeta3-mediated fibrino-gen adhesion. In cell experiments during TGFbeta-induced epithelial to Disease mesenchymal transdifferen-tiation (EMT), Dab2 DAB2 expression is down-regulated. expression is increased and Dab2 binds to beta1

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 366 DAB2 (disabled homolog 2, mitogen-responsive phosphoprotein (Drosophila)) Orlandini M

Malignant peripheral nerve sheath structure formation during mouse embryogenesis. Dev Biol. 2002 Nov 1;251(1):27-44 tumor, invasive cervical carcinoma He J, Xu J, Xu XX, Hall RA. Cell cycle-dependent Note phosphorylation of Disabled-2 by cdc2. Oncogene. 2003 Jul Comparative genomic hybridization (CGH) revealed 17;22(29):4524-30 frequent gains of DAB2. Zhou J, Scholes J, Hsieh JT. Characterization of a novel negative regulator (DOC-2/DAB2) of c-Src in normal prostatic References epithelium and cancer. J Biol Chem. 2003 Feb 28;278(9):6936- 41 Xu XX, Yang W, Jackowski S, Rock CO. Cloning of a novel Huang CL, Cheng JC, Liao CH, Stern A, Hsieh JT, et al. phosphoprotein regulated by colony-stimulating factor 1 shares Disabled-2 is a negative regulator of integrin alpha(IIb)beta(3)- a domain with the Drosophila disabled gene product. J Biol mediated fibrinogen adhesion and cell signaling. J Biol Chem. Chem. 1995 Jun 9;270(23):14184-91 2004 Oct 1;279(40):42279-89 Albertsen HM, Smith SA, Melis R, Williams B, Holik P, Stevens Hidalgo A, Baudis M, Petersen I, Arreola H, Piña P, et al. J, White R. Sequence, genomic structure, and chromosomal Microarray comparative genomic hybridization detection of assignment of human DOC-2. Genomics. 1996 Apr chromosomal imbalances in uterine cervix carcinoma. BMC 15;33(2):207-13 Cancer. 2005 Jul 9;5:77 Mok SC, Chan WY, Wong KK, Cheung KK, Lau CC, Ng SW, Prunier C, Howe PH. Disabled-2 (Dab2) is required for Baldini A, Colitti CV, Rock CO, Berkowitz RS. DOC-2, a transforming growth factor beta-induced epithelial to candidate tumor suppressor gene in human epithelial ovarian mesenchymal transition (EMT). J Biol Chem. 2005 Apr cancer. Oncogene. 1998 May 7;16(18):2381-7 29;280(17):17540-8 Fazili Z, Sun W, Mittelstaedt S, Cohen C, Xu XX. Disabled-2 Zhoul J, Hernandez G, Tu SW, Huang CL, Tseng CP, Hsieh inactivation is an early step in ovarian tumorigenicity. JT. The role of DOC-2/DAB2 in modulating androgen receptor- Oncogene. 1999 May 20;18(20):3104-13 mediated cell growth via the nongenomic c-Src-mediated Tseng CP, Ely BD, Pong RC, Wang Z, Zhou J, Hsieh JT. The pathway in normal prostatic epithelium and cancer. Cancer role of DOC-2/DAB2 protein phosphorylation in the inhibition of Res. 2005 Nov 1;65(21):9906-13 AP-1 activity. An underlying mechanism of its tumor- Nakagawa Y, Yoshida A, Numoto K, Kunisada T, Wai D, Ohata suppressive function in prostate cancer. J Biol Chem. 1999 N, Takeda K, Kawai A, Ozaki T. Chromosomal imbalances in Nov 5;274(45):31981-6 malignant peripheral nerve sheath tumor detected by He J, Smith ER, Xu XX. Disabled-2 exerts its tumor suppressor metaphase and microarray comparative genomic hybridization. activity by uncoupling c-Fos expression and MAP kinase Oncol Rep. 2006 Feb;15(2):297-303 activation. J Biol Chem. 2001 Jul 20;276(29):26814-8 Zhou J, Fan J, Hsieh JT. Inhibition of mitogen-elicited signal Hocevar BA, Smine A, Xu XX, Howe PH. The adaptor transduction and growth in prostate cancer with a small peptide molecule Disabled-2 links the transforming growth factor beta derived from the functional domain of DOC-2/DAB2 delivered receptors to the Smad pathway. EMBO J. 2001 Jun by a unique vehicle. Cancer Res. 2006 Sep 15;66(18):8954-8 1;20(11):2789-801 Karam JA, Shariat SF, Huang HY, Pong RC, Ashfaq R, et al. Morris SM, Cooper JA. Disabled-2 colocalizes with the LDLR in Decreased DOC-2/DAB2 expression in urothelial carcinoma of clathrin-coated pits and interacts with AP-2. Traffic. 2001 the bladder. Clin Cancer Res. 2007 Aug 1;13(15 Pt 1):4400-6 Feb;2(2):111-23 Spudich G, Chibalina MV, Au JS, Arden SD, Buss F, Kendrick- Smith ER, Capo-chichi CD, He J, Smedberg JL, et al. Jones J. Myosin VI targeting to clathrin-coated structures and Disabled-2 mediates c-Fos suppression and the cell growth dimerization is mediated by binding to Disabled-2 and regulatory activity of retinoic acid in embryonic carcinoma cells. PtdIns(4,5)P2. Nat Cell Biol. 2007 Feb;9(2):176-83 J Biol Chem. 2001 Dec 14;276(50):47303-10 Yang DH, Cai KQ, Roland IH, Smith ER, Xu XX. Disabled-2 is Wang SC, Makino K, Xia W, Kim JS, Im SA, Peng H, Mok SC, an epithelial surface positioning gene. J Biol Chem. 2007 Apr Singletary SE, Hung MC. DOC-2/hDab-2 inhibits ILK activity 27;282(17):13114-22 and induces anoikis in breast cancer cells through an Akt- Jiang Y, Prunier C, Howe PH. The inhibitory effects of independent pathway. Oncogene. 2001 Oct 18;20(47):6960-4 Disabled-2 (Dab2) on Wnt signaling are mediated through Zhou J, Hsieh JT. The inhibitory role of DOC-2/DAB2 in growth Axin. Oncogene. 2008 Mar 20;27(13):1865-75 factor receptor-mediated signal cascade. DOC-2/DAB2- Orlandini M, Nucciotti S, Galvagni F, Bardelli M, et al. mediated inhibition of ERK phosphorylation via binding to Morphogenesis of human endothelial cells is inhibited by DAB2 Grb2. J Biol Chem. 2001 Jul 27;276(30):27793-8 via Src. FEBS Lett. 2008 Jul 23;582(17):2542-8 Mishra SK, Keyel PA, Hawryluk MJ, Agostinelli NR, Watkins Yang DH, Smith ER, Cai KQ, Xu XX. C-Fos elimination SC, Traub LM. Disabled-2 exhibits the properties of a cargo- compensates for disabled-2 requirement in mouse selective endocytic clathrin adaptor. EMBO J. 2002 Sep extraembryonic endoderm development. Dev Dyn. 2009 16;21(18):4915-26 Mar;238(3):514-23 Rosenbauer F, Kallies A, Scheller M, Knobeloch KP, et al Disabled-2 is transcriptionally regulated by ICSBP and This article should be referenced as such: augments macrophage spreading and adhesion. EMBO J. Orlandini M. DAB2 (disabled homolog 2, mitogen-responsive 2002 Feb 1;21(3):211-20 phosphoprotein (Drosophila)). Atlas Genet Cytogenet Oncol Yang DH, Smith ER, Roland IH, Sheng Z, He J, et al. Haematol. 2010; 14(4):365-367. Disabled-2 is essential for endodermal cell positioning and

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Gene Section Review

DLG1 (discs, large homolog 1 (Drosophila)) Paola Massimi, Lawrence Banks International Centre for Genetic Engeneering and Biotechnology (ICGEB), Trieste, Italy (PM, LB)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/DLG1ID40333ch3q29.html DOI: 10.4267/2042/44730 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

possess a class I PDZ binding motif (Morais Cabral et Identity al., 1996). Other names: DKFZp761P0818; DKFZp781B0426; There are two major transcripts of DLG1 gene. One is DLGH1; SAP-97; SAP97; dJ1061C18.1.1; Hdlg Discs large homolog 1 isoform 1, which contains an HGNC (Hugo): DLG1 additional exon (99 nucleotides) in the 5' part of the Location: 3q29 Dlg homology repeats (DHR) domain and lacks an exon in the 3' coding region, resulting in a shorter DNA/RNA protein (isoform 1), compared to isoform 2. The second is Discs large homolog 1 isoform 2, which represents Description the longer transcript and encodes the longer isoform. This second transcript is alternatively spliced with an The DLG1 gene consists of 250,017 bases on the 3q29 insertion of 34 nucleo-tides in the region between the locus of chromosome 3 (Azim et al., 1995). SH3 and GUK (isoform 2). Another alternative splice Transcription has an inser-tion of 100 nucleotides and the resulting The DLG1 gene encodes a 960 amino-acid protein of transcript is called Discs large homolog 1 isoform 3. 100355 Da with several distinct domains. A 1310-bp In conclusion, the protein is regulated by a several fragment of the 5' flanking region of the DLG1 gene, different alternative splicing events (Mori et al., 1998) corresponding to nucleotide (nt) - 1217/+ 93 contains resulting in a number of different combination of the promoter sequence plus the consensus-binding sites spliced variants (which give raise to at least 7 isoforms for the Snail family of transcription factors that repress such as I1I2, I1I3, etc.) (see table), some of which are the expression of some epithelial markers and are up- transcribed in a tissue-specific manner (Lue et al., regulated in a variety of tumours. Snail transcription 1996; McLaughlin et al., 2002). factors repress the transcriptional activity of the DLG1 Pseudogene promoter (Cavatorta et al., 2008). The carboxyl- terminal 179 aa show strong homology (35.5%) to None. yeast guanylate kinase (GUK) an enzyme that transfers a phosphate group from ATP to GMP, converting it to Protein GDP, although DLG1 has no enzymatic activity. Description DLG1 contains also a 59 aa SH3 (Src homolgy 3) domain, which mainly mediates binding to other The 'discs large' protein, Dlg1, is part of a family of proteins. The N terminal half of the molecule contains proteins termed MAGUKs (membrane-associated three copies of the 80-90 aa motif called guanylate kinase homologs). MAGUKs are loca-lized DHR/GLGF/PDZ (PSD-95, Dlg, ZO-1), which mediate at the membrane-cytoskeleton interface, usually at cell- the binding of the protein to the plasma membrane and cell junctions, where they appear to have both confers binding to proteins that structural and signaling roles. DLG1 probably exists as an homotetramer.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 368 DLG1 (discs, large homolog 1 (Drosophila)) Massimi P, Banks L

Diagram of DLG1 gene organization and of the two major encoded transcript variants.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 369 DLG1 (discs, large homolog 1 (Drosophila)) Massimi P, Banks L

Diagram of the DLG1 protein with its characteristic domains and the main protein-protein interactions at the cell-cell junctions.

Ultrastructural analysis of hDlg by low angle rotary al., 1996; Bonilha and Rodriguez-Boulan, 2001). shadow electron microscopy revealed that the full- Indeed SAP97 also interacts with ezrin, an actin- length hDlg protein as well as its amino-terminal binding protein crucial for morphogenesis of apical domain exhibits a highly flexible irregular shape. microvilli and basolateral infoldings in retinal pigment Further evaluation of the self-association state of hDlg epithelial (RPE) cells. using sedimentation equilibrium centrifugation, matrix- Through the PDZ2 domain the protein also interacts assisted laser desorption/ionization mass spectrometry with the carboxyl-terminal S/TXV motif of the APC and chemical cross-linking techniques confirmed that (Adenomatous polyposis coli) tumour suppressor the oligomerization site of hDlg is contained within its protein and plays an important role in transducing the amino-terminal domain. This is mediated by a unique APC cell cycle blocking signal (Makino et al., 1997; L27 domain which regulates multimerization of hDlg Ishidate et al., 2000; Mimori-Kiyosue et al., 2007). In into dimeric and tetrameric species in solution, and addition, APC appears to mediate the interaction sedimentation velocity experiments demonstrated that between DLG1, beta-catenin and the actin the oligomerization domain exists as an elongated cytoskeleton. Beta-catenin is complexed with gamma- tetramer in solution (Marfatia et al., 2000). Thus, the catenin and alpha-catenin, through which DLG1 binds L27 domain regulate DLG1 self-association. The N- to E-cadherin (Reuver et al., 1998). Moreover, the Src terminal alternatively spliced region is capable of homology domain 2 of the p85/PI3K and hDlg are binding several SH3 domains and also moderates the associated with E-cadherin in a common level of protein oligomerization. macromolecular complex in differentiating intestinal Specific binding partners are known for each domain of cells, and in this way hDlg may be a determinant in E- DLG1, and different modes of intramolecular cadherin-mediated adhesion and signaling in interactions have been proposed that particularly mammalian epithelial cells (Laprise et al., 2004). involve the SH3 and GUK domains and the so-called DLG1 was demonstrated also to bind with voltage- HOOK region located between these two domains. gated or Kv K(+) channels through its PDZ domains DLG1 binds to the membrane cytoskeletal 4.1 protein (Hanada et al., 1997; Tiffany et al., 2000; Eldstrom et through its C-terminal region (Hanada et al., 2003), via al., 2003). The complex formation involves the a motif encoded by the alternatively spliced exon association of Cav-3 with a segment of SAP97 located between the SH3 and the C-terminal guanylate localized between its PDZ2 and PDZ3 domains. This kinase-like domains (Isoform I3). The PDZ1-2 modules scaffolding complex can recruit Kv1.5 to form a and the I3 domain associate with the 30-kD NH2- tripartite complex in which each of the three terminal domain of protein 4.1 that is conserved in components interacts with the other two. These ezrin/radixin/moesin (ERM) proteins module (Lue et

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interactions between Kv1.5, Cav3 and SAP97 may (Hanada et al., 2000; Yamada et al., 2007; Unno et al., constitute the nucleation site for the assembly of 2008). macromolecular containing potassium channels and Using the yeast two-hybrid screening a novel protein thereby regulates cellular potential currents (Folco et from a human cDNA library was isolated as a binding al., 2004). partner of DLG1. This protein is a component of TJs Hanada showed by immunoblot analysis that rather than AJs (where DLG1 is normally found), even immunoprecipitates of DLG1 in T lymphocytes contain if it is incorporated into TJs after TJ strands are formed, the Src family tyrosine kinase p56 (lck). Binding and therefore it is named Pilt (protein incorporated later analysis demonstrated that LCK interacts with the into TJs) (Kawabe et al., 2001). proline-rich N-terminal domain of DLG1, suggesting DLG1 is known to interact also with several human that DLG1 may function as a coupler of tyrosine kinase virus oncoproteins : HPV E6 (Lee et al., 1997; Kiyono and a voltage-gated potassium channel in T et al.,1997, Gardiol et al., 1999) through its C-terminus lymphocytes. and DLG1 PDZ2 domain and as result is subjected to The HOOK region of DLG1 is also a specific site for proteasome mediated degradation; HTLV-1 TAX calmodulin binding and interaction of SAP97 to (Suzuki et al., 1999), via the C-terminus of Tax and the immobilized calmodulin is strictly calcium-depen-dent PDZ domain of hDLG. Tax prevents the binding of (Paarmann et al., 2002). The calmodulin seems to hDLG to APC tumor suppressor gene product, regulate the intramolecular interaction between the suggesting the mechanism for inhibition of hDLG SH3, HOOK, and GK domains of the protein. function; Adenovirus type 9 E4-ORF1 specifically DLG1 also forms multiprotein complexes with CASK, requires endogenous DLG1 to provoke oncogenic LIN7A, LIN7B, LIN7C, APBA1, and KCNJ12 (Nix et activation of phosphatidyl-inositol 3-kinase (PI3K) in al., 2000; Lee et al., 2002; Leonoudakis et al., 2004) cells. E4-ORF1 binding to Dlg1 on ts PDZ domain and exists as a tripartite complex composed of DLG1, triggers the resulting complex to translocate to the MPP7 and LIN7 (LIN7A or LIN7C) (Bohl et al., 2007; plasma membrane and, at this site, to promote Ras- Stucke et al., 2007). MPP7 dimerized with the LIN7 mediated PI3K activation, suggesting a surprising proteins through its L27C domain. The LIN7/MPP7 oncogenic function for DLG1 in virus-mediated dimer then linked to DLG1 though the L27N domain of cellular transformation (Frese et al., 2006; Chung et al., MPP7. This complex localizes to epithelial adherens 2007). junctions in transfected Madin-Darby Canine Kidney hDlg also binds the tumor endothelial marker 5 cells (MDCK). MPP7 constructs lacking either the PDZ (TEM5), a seven-pass transmembrane protein that is or SH3 domain redistributed MPP7, DLG1, and LIN7 homologous to the B family of G-protein-coupled into the soluble cytoplasmic fraction. MPP7 and DLG1 receptors (GPCRs). The PDZ domains of hDlg bound colocalized at the lateral surface of epithelial cells, and the C-terminal PDZ-binding motif of TEM5. DLG1 is they overlapped with markers of adherens junctions furthermore able to interact with a novel seven-pass and tight junctions. Loss of either DLG1 or MPP7 from transmembrane protein, which was homologous to epithelial cells resulted in a significant defect in TEM5, and was named here a TEM5-like protein assembly and maintenance of functional tight junctions. (TEM5-like) (Yamamoto et al., 2004). The formation of the DLG1-MPP7 complex promotes SAP97/hDlg as a scaffold protein is also targeted to the also epithelial cell polarity. cytoskeleton by its association with the protein SAP97 binds two other mLIN-7 binding MAGUK guanylate kinase-associated protein (GKAP), which is proteins. One of these MAGUK proteins, DLG3, part of the postsynaptic scaffold in neuronal cells coimmunoprecipitates with SAP97 in lysates from rat (Sabio et al., 2005). Moreover, hDlg is believed to brain and transfected MDCK cells. This interaction associate with AMPA receptors (AMPARs) containing requires the MRE (MAGUK recruitment) domain of the GluR1 subunit, but the functional significance of SAP97 and surprisingly, both the L27N and L27 these interactions is partially unclear, even if this carboxyl-terminal (L27C) domains of DLG3. SAP97 interaction seems to be occur early in the secretory can interact with the MAGUK protein, DLG2, but not pathway, while the receptors are in the endoplasmic the highly related protein, PALS2. The ability of reticulum or cis-Golgi (Sans et al., 2001). In light SAP97 to interact with multiple MAGUK proteins is membrane fractions prepared from rat brain, myosin VI likely to be important for the targeting of specific and SAP97 form a trimeric complex with the alpha- protein complexes in polarized cells (Karnak et al., amino-3-hydroxy-5-methylisoxazole-4-propionic acid 2002). (AMPA) receptor subunit, GluR1. It is possible that The kinesin-3 motor protein, GAKIN, is regulated by SAP97 may serve as a molecular link between GluR1 the direct binding of its protein cargo hDlg. Direct and the actin-dependent motor protein myosin VI binding of the SH3-I3-GUK module of hDlg to the during the dynamic translocation of AMPA receptors to MAGUK Binding Stalk domain of GAKIN activates and from the postsynaptic plasma membrane (Wu et al., the microtubule-stimulated ATPase activity of GAKIN 2002).

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DLG1 is also able to translocate to the immune synapse et al., 2007). It is also found at the immunological and lipid rafts in response to T cell receptor synapse, endoplasmic reticulum, endoplasmic (TCR)/CD28 engagement and LckSH3-mediated reticulum membrane, postsynaptic density, lateral interactions with DLG1 control its membrane targeting. plasma membrane, neuromuscular junction membrane, TCR/CD28 engagement induces the formation of raft synapse and the post-synaptic membrane. There is endogenous Lck-DLG1-Zap70-Wiskott-Aldrich equal expression of the two spliced variants in most syndrome protein (WASp) complexes in which DLG1 human tissues; however, in skeletal muscle the acts to facilitate interactions of Lck with Zap70 and transcript with the 99-bp insertion is predominant, WASp (Round et al., 2005). whilst in the brain, the isoform lacking the 99-bp Delta 1 acts as a membrane-bound ligand that interacts insertion is predominant. In brain there are six with the Notch receptor and plays a critical role in cell different, alternatively spliced transcripts, two of which fate specification. DLG1 binds the Delta 1 C-terminal included a novel, 36-bp, brain-specific exon encoding a region, in a PDZ dependent manner. Delta 4 also peptide bearing significant homology to a portion of rat interacts with DLG1, whereas Jagged1, another Notch synapse-associated proteins, SAP97 and PSD95. ligand, does not (Six et al., 2004). Again, the different isoforms of the protein seem to MARCH 2, which is part of the MARCH family have diverse localisation in the cell. I2 and I3 variants ubiquitin ligases and is implicated in the endosomal have distinct distributions in epidermal and cervical trafficking interacts with full-length DLG1 in a PDZ epithelia. In skin and cervix, I3 variants are found in domain dependent manner. Furthermore, MARCH2 co- the cytoplasm. Cytoplasmic localization of I3 variants localized with DLG1 at sites of cell-cell contact (Cao et decreases as cervical keratinocytes differentiate, al., 2008). concomitant with relocalization to the cell periphery. I2 variants are found at the cell periphery of differentiated SAP97 is a binding partner of the cytoplasmic domain epidermal and cervical keratinocytes. Nuclear of TACE, which is the Tumour necrosis factor alpha localization of I2 variants is evident in both tissues, converting enzyme and is the metalloprotease- with a concentration of nuclear I2 variants in basal and disintegrin responsible for the ectodomain shedding of parabasal cervical keratinocytes (Roberts et al., 2007), several proteins, including tumour necrosis factor underlining that different hDlg isoforms play distinct alpha. The interaction involved the PDZ3 domain of roles at various stages of epithelial differentiation. SAP97 and the extreme C-terminal amino-acid More-over, upon transient transfection into sequence of TACE (Peiretti et al., 2003). subconfluent (MDCK) epithelial cells, hDlg-I3 DLG1 is able to interact also with Net 1 which is a accumulated predominantly at the plasma membrane of nuclear RhoA-specific guanine nucleotide exchange cell-cell contact sites, whereas hDlg-I2 distributed in factor. The binding is through the PDZ-binding motif. the cytoplasm. The hDlg-I3 but not the hDlg-I2 isoform The ability of oncogenic Net1 to transform cells may binds to the FERM (Four.1-Ezrin-Radixin-Moesin) be in part related to its ability to sequester tumour domain of protein 4.1, playing a critical role in suppressor proteins like DLG1 in the cytosol, thereby recruiting DLG1 to the lateral membrane in epithelial interfering with their normal cellular function (Garcia- cells (Bonhila et al., 2001; Hanada et al., 2003; Mata et al., 2007). Massimi et al., 2003; Wu et al., 2002). DLG1 interacts with the tSNARE syntaxin 4 which is Several different domains of DLG1 contribute to its involved in vesicle transport, and this binding may localisation. Mutation of the SH3 or GUK domain, but contribute to the correct colocaliation of the other not the PDZ domain, results in a re-localization of proteins of the Scrib complex: hScribble and Hugl-1 hDLG to the nucleus and, moreover, DLG1 possess a (Massimi et al., 2008). potential nuclear localization signal in the HOOK Expression domain (Kohu et al., 2002). It has been reported that the localisation of DLG1 is DLG1 is widely expressed, with different isoforms also dependent on the post-translational modifica-tion displaying different expression profiles (McLaughlin et of the protein, by phosphorylation occurring post- al., 2002). DLG1 is expressed mainly in epithelial cells osmotic shock (Massimi et al., 2006) and also during and in the nervous system, but is also fond in thymus, the cell cycle following CDK phosphoryla-tion bone marrow, T cells, spleen, brain, spinal cord, heart, (Narayan et al., 2009). Moreover, DLG1 localises kidney, lung, liver, dependently from the other proteins involved in the pancreas, prostate (at the protein level). complex at the adherens junctions: Localisation hScribble and Hugl-1 (Massimi et al., 2008). DLG1 is localised at the plasma membrane (Hanada et In addition, CaMKII (calcium/calmodulin-depen-dent al., 2000), cell-cell junctions (Lue et al., 1994), at the protein kinase II) activation led to increased targeting basolateral plasma membrane (Lue et al., 1996; Mimori of SAP97 into dendritic spines, whereas CaMKII

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inhibition was responsible for SAP97 colocalization in the cell soma with the endoplasm-mic reticulum protein Mutations disulfide-isomerase (Mauceri et al., 2004). Note Regarding the localisation of the different isoforms, the See paragraphs below. two main cardiac SAP97 isoforms contains both I3 and Germinal I1B inserts and differs by the I1A insert. Both isoforms co-precipitate with hKv1.5 channels, and have different None. effects on the hKv1.5 current, depending on their Somatic capacity to form clusters (Godreau et al., 2003). There is one report in breast cancer (Fuja et al., 2004). In the case of endothelial cells of embryonic liver the expression of TEM5 colocalises with DLG1. This Implicated in suggest that hDlg localizes at the plasma membrane through TEM5 and TEM5-like proteins and Epithelial-derived cancers furthermore scaffolds these GPCRs in endothelial cells Note during tumour angiogenesis and neoangiogenesis The mis-localisation of DLG1 is linked to the (Yamamoto et al., 2004). development of epithelial-derived cancers (Gardiol et Function al., 2006). In uterine cervical squamous epithelia, DLG1 is an essential multidomain scaffolding protein prominent localization of hDlg at sites of inter-cellular required for normal development. contact occurs in cells that have left the proliferating basal cell layers and begun maturation. The presence of It is able to recruits channels (Hanada et al., 1997; hDlg at sites of cell-cell contact diminishes, whilst Tiffany et al., 2000; Abi-Char et al., 2008), receptors intracellular cytoplasmic levels increase significantly in and signaling molecules (Sans et al., 2001; Wuh et al., high-grade, but not low-grade, cervical neoplasias. In 2002; Six et al., 2004) to discrete plasma membrane invasive squamous cell carcinomas, total cellular hDlg domains in polarized cells. Its main role is played in levels are greatly reduced (Watson et al., 2002). adherens junctions assembly (Laprise et al., 2004; Bohl et al., 2007; Stucke et al., 2007; Massimi et al., 2008). Mammary ductal carcinoma However DLG1 with the establishment of a Note multiprotein complexes at cell-cell contacts is also In humans there is only one report of mutations involved in signal transduction (Massimi et al., 2006), occuring in Dlg in cancer. In this study somatic cell prolifera-tion (Suzuki et al., 1999; Ishidate et al., mutations were found in three genes (CSNK1 epsilon, 2000; Massimi et al., 2003; Thomas et al., 2005; Frese encoding the Ser/Thr kinase casein kinase I epsilon, et al., 2006; Garcia-Mata et al., 2007; Unno et al., DLG1, and EDD/hHYD, encoding a pro-gestin induced 2008), synaptogenesis (Mori et al., 1998; Sans et al., putative ubiquitin-protein ligase) in mammary ductal 2001; Mauceri et al., 2004), lymphocyte activation carcinoma. For CSNK1 epsilon and DLG1, most of the (Hanada et al., 1997; Hanada et al., 2000; Round et al., mutations affected highly conserved residues, some 2005), cell differentiation (Laprise et al., 2004; Roberts were found repetitively in different patients, and no et al., 2007), cell migration (Six et al., 2004) and synonymous mutations were found, indicating that the cellular apical-basal polarity control (Bonilha et al., observed mutations were selected in tumours and may 2001). be functionally significant (Fuja et al., 2004). Homology 3q29 microdeletion syndrome The four best-characterised mammalian Dlg family Note members are Dlg1 (hDlg/SAP97), Dlg2 (PSD- Moreover, another report (Willatt et al., 2005) pointed 93/Chapsyn-110), Dlg3 (NE-Dlg/SAP102) and Dlg4 out that the DLG1 and PAK2 genes are deleted in the (PSD-95/SAP90) (Lue et al., 1994; Makno et al., 1997; 3q29 microdeletion syndrome and raised the possibility Brenman et al., 1996; Cho et al., 1992; Humbert et al., that loss of one of these genes may contribute to the 2003). Mammalian Dlg family members display the phenotype since PAK2 and DLG1 are autosomal characteristic MAGUK structural domains found in homologs of 2 X-linked mental retardation genes, Drosophila Dlg including the three PDZ domains, a Src PAK3 and DLG3. homology domain-3 (SH3) and a guanylate kinase-like Schizophrenia (GUK) domain. Although most mammalian Dlg homologues were first identified in neuronal tissues, all Note of these proteins are expressed in a variety of non- In addition, DLG1 gene may be a susceptibility factor neuronal tissues including epithelial and lymphoid in male schizophrenics and the modification of the cells. Strikingly, localisation studies in all of these glutamate receptor signalling pathway could be tissues are suggestive of a role for mammalian Dlg involved in the disease pathophysiology. DLG1 protein homologues in polarisation. levels were decreased to less than half that of the

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control levels specifically in the prefrontal cortex of Mori K, Iwao K, Miyoshi Y, Nakagawara A, Kofu K, Akiyama T, schizophrenic patients. In parallel, its binding partner, Arita N, Hayakawa T, Nakamura Y. Identification of brain- specific splicing variants of the hDLG1 gene and altered GluR1, similarly decreased in the same brain region splicing in neuroblastoma cell lines. J Hum Genet. (Toyooka et al., 2002; Sato et al., 2008). 1998;43(2):123-7 Various cancer Reuver SM, Garner CC. E-cadherin mediated cell adhesion recruits SAP97 into the cortical cytoskeleton. J Cell Sci. 1998 Note Apr;111 ( Pt 8):1071-80 Generally, loss of expression (through diverse Wu H, Reuver SM, Kuhlendahl S, Chung WJ, Garner CC. mechanisms) is a common feature in many late stages Subcellular targeting and cytoskeletal attachment of SAP97 to of cancers. the epithelial lateral membrane. J Cell Sci. 1998 Aug;111 ( Pt 16):2365-76 References Gardiol D, Kühne C, Glaunsinger B, Lee SS, Javier R, Banks L. 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Lee S, Fan S, Makarova O, Straight S, Margolis B. A novel and Mauceri D, Cattabeni F, Di Luca M, Gardoni F. conserved protein-protein interaction domain of mammalian Calcium/calmodulin-dependent protein kinase II Lin-2/CASK binds and recruits SAP97 to the lateral surface of phosphorylation drives synapse-associated protein 97 into epithelia. Mol Cell Biol. 2002 Mar;22(6):1778-91 spines. J Biol Chem. 2004 May 28;279(22):23813-21 McLaughlin M, Hale R, Ellston D, Gaudet S, Lue RA, Viel A. Six EM, Ndiaye D, Sauer G, Laâbi Y, Athman R, Cumano A, The distribution and function of alternatively spliced insertions Brou C, Israël A, Logeat F. The notch ligand Delta1 recruits in hDlg. J Biol Chem. 2002 Feb 22;277(8):6406-12 Dlg1 at cell-cell contacts and regulates cell migration. J Biol Chem. 2004 Dec 31;279(53):55818-26 Paarmann I, Spangenberg O, Lavie A, Konrad M. Formation of complexes between Ca2+.calmodulin and the synapse- Yamamoto Y, Irie K, Asada M, Mino A, Mandai K, Takai Y. associated protein SAP97 requires the SH3 domain-guanylate Direct binding of the human homologue of the Drosophila disc kinase domain-connecting HOOK region. J Biol Chem. 2002 large tumor suppressor gene to seven-pass transmembrane Oct 25;277(43):40832-8 proteins, tumor endothelial marker 5 (TEM5), and a novel TEM5-like protein. Oncogene. 2004 May 13;23(22):3889-97 Toyooka K, Iritani S, Makifuchi T, Shirakawa O, Kitamura N, Maeda K, Nakamura R, Niizato K, Watanabe M, Kakita A, Round JL, Tomassian T, Zhang M, Patel V, Schoenberger SP, Takahashi H, Someya T, Nawa H. Selective reduction of a Miceli MC. Dlgh1 coordinates actin polymerization, synaptic T PDZ protein, SAP-97, in the prefrontal cortex of patients with cell receptor and lipid raft aggregation, and effector function in chronic schizophrenia. J Neurochem. 2002 Nov;83(4):797-806 T cells. J Exp Med. 2005 Feb 7;201(3):419-30 Watson RA, Rollason TP, Reynolds GM, Murray PG, Banks L, Sabio G, Arthur JS, Kuma Y, Peggie M, Carr J, Murray-Tait V, Roberts S. Changes in expression of the human homologue of Centeno F, Goedert M, Morrice NA, Cuenda A. p38gamma the Drosophila discs large tumour suppressor protein in high- regulates the localisation of SAP97 in the cytoskeleton by grade premalignant cervical neoplasias. Carcinogenesis. 2002 modulating its interaction with GKAP. EMBO J. 2005 Mar Nov;23(11):1791-6 23;24(6):1134-45 Wu H, Nash JE, Zamorano P, Garner CC. Interaction of Thomas M, Massimi P, Navarro C, Borg JP, Banks L. The SAP97 with minus-end-directed actin motor myosin VI. hScrib/Dlg apico-basal control complex is differentially targeted Implications for AMPA receptor trafficking. J Biol Chem. 2002 by HPV-16 and HPV-18 E6 proteins. Oncogene. 2005 Sep Aug 23;277(34):30928-34 15;24(41):6222-30 Eldstrom J, Choi WS, Steele DF, Fedida D. SAP97 increases Willatt L, Cox J, Barber J, Cabanas ED, Collins A, Donnai D, Kv1.5 currents through an indirect N-terminal mechanism. FitzPatrick DR, Maher E, Martin H, Parnau J, Pindar L, FEBS Lett. 2003 Jul 17;547(1-3):205-11 Ramsay J, Shaw-Smith C, Sistermans EA, Tettenborn M, Trump D, de Vries BB, Walker K, Raymond FL. 3q29 Godreau D, Vranckx R, Maguy A, Goyenvalle C, Hatem SN. microdeletion syndrome: clinical and molecular Different isoforms of synapse-associated protein, SAP97, are characterization of a new syndrome. Am J Hum Genet. 2005 expressed in the heart and have distinct effects on the voltage- Jul;77(1):154-60 gated K+ channel Kv1.5. J Biol Chem. 2003 Nov 21;278(47):47046-52 Frese KK, Latorre IJ, Chung SH, Caruana G, Bernstein A, Jones SN, Donehower LA, Justice MJ, Garner CC, Javier RT. Hanada T, Takeuchi A, Sondarva G, Chishti AH. Protein 4.1- Oncogenic function for the Dlg1 mammalian homolog of the mediated membrane targeting of human discs large in Drosophila discs-large tumor suppressor. EMBO J. 2006 Mar epithelial cells. J Biol Chem. 2003 Sep 5;278(36):34445-50 22;25(6):1406-17 Humbert P, Russell S, Richardson H. Dlg, Scribble and Lgl in Gardiol D, Zacchi A, Petrera F, Stanta G, Banks L. Human cell polarity, cell proliferation and cancer. Bioessays. 2003 discs large and scrib are localized at the same regions in colon Jun;25(6):542-53 mucosa and changes in their expression patterns are Massimi P, Gardiol D, Roberts S, Banks L. Redistribution of correlated with loss of tissue architecture during malignant the discs large tumor suppressor protein during mitosis. Exp progression. Int J Cancer. 2006 Sep 15;119(6):1285-90 Cell Res. 2003 Nov 1;290(2):265-74 Massimi P, Narayan N, Cuenda A, Banks L. Phosphorylation of Peiretti F, Deprez-Beauclair P, Bonardo B, Aubert H, Juhan- the discs large tumour suppressor protein controls its Vague I, Nalbone G. Identification of SAP97 as an intracellular membrane localisation and enhances its susceptibility to HPV binding partner of TACE. J Cell Sci. 2003 May 15;116(Pt E6-induced degradation. Oncogene. 2006 Jul 20;25(31):4276- 10):1949-57 85 Folco EJ, Liu GX, Koren G. Caveolin-3 and SAP97 form a Bohl J, Brimer N, Lyons C, Vande Pol SB. The stardust family scaffolding protein complex that regulates the voltage-gated protein MPP7 forms a tripartite complex with LIN7 and DLG1 potassium channel Kv1.5. Am J Physiol Heart Circ Physiol. that regulates the stability and localization of DLG1 to cell 2004 Aug;287(2):H681-90 junctions. J Biol Chem. 2007 Mar 30;282(13):9392-400 Fuja TJ, Lin F, Osann KE, Bryant PJ. Somatic mutations and Chung SH, Frese KK, Weiss RS, Prasad BV, Javier RT. A new altered expression of the candidate tumor suppressors CSNK1 crucial protein interaction element that targets the adenovirus epsilon, DLG1, and EDD/hHYD in mammary ductal carcinoma. E4-ORF1 oncoprotein to membrane vesicles. J Virol. 2007 Cancer Res. 2004 Feb 1;64(3):942-51 May;81(9):4787-97 Laprise P, Viel A, Rivard N. Human homolog of disc-large is García-Mata R, Dubash AD, Sharek L, Carr HS, Frost JA, required for adherens junction assembly and differentiation of Burridge K. The nuclear RhoA exchange factor Net1 interacts human intestinal epithelial cells. J Biol Chem. 2004 Mar with proteins of the Dlg family, affects their localization, and 12;279(11):10157-66 influences their tumor suppressor activity. Mol Cell Biol. 2007 Dec;27(24):8683-97 Leonoudakis D, Conti LR, Radeke CM, McGuire LM, Vandenberg CA. A multiprotein trafficking complex composed Mimori-Kiyosue Y, Matsui C, Sasaki H, Tsukita S. of SAP97, CASK, Veli, and Mint1 is associated with inward Adenomatous polyposis coli (APC) protein regulates epithelial rectifier Kir2 potassium channels. J Biol Chem. 2004 Apr cell migration and morphogenesis via PDZ domain-based 30;279(18):19051-63

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 375 DLG1 (discs, large homolog 1 (Drosophila)) Massimi P, Banks L

interactions with plasma membranes. Genes Cells. 2007 Cavatorta AL, Giri AA, Banks L, Gardiol D. Cloning and Feb;12(2):219-33 functional analysis of the promoter region of the human Disc large gene. Gene. 2008 Nov 15;424(1-2):87-95 Roberts S, Calautti E, Vanderweil S, Nguyen HO, Foley A, Baden HP, Viel A. Changes in localization of human discs Massimi P, Narayan N, Thomas M, Gammoh N, Strand S, large (hDlg) during keratinocyte differentiation are [corrected] Strand D, Banks L. Regulation of the hDlg/hScrib/Hugl-1 associated with expression of alternatively spliced hDlg tumour suppressor complex. Exp Cell Res. 2008 Nov variants. Exp Cell Res. 2007 Jul 15;313(12):2521-30 1;314(18):3306-17 Stucke VM, Timmerman E, Vandekerckhove J, Gevaert K, Hall Sato J, Shimazu D, Yamamoto N, Nishikawa T. An association A. The MAGUK protein MPP7 binds to the polarity protein analysis of synapse-associated protein 97 (SAP97) gene in hDlg1 and facilitates epithelial tight junction formation. Mol Biol schizophrenia. J Neural Transm. 2008 Sep;115(9):1355-65 Cell. 2007 May;18(5):1744-55 Unno K, Hanada T, Chishti AH. Functional involvement of Yamada KH, Hanada T, Chishti AH. The effector domain of human discs large tumor suppressor in cytokinesis. Exp Cell human Dlg tumor suppressor acts as a switch that relieves Res. 2008 Oct 15;314(17):3118-29 autoinhibition of kinesin-3 motor GAKIN/KIF13B. Biochemistry. 2007 Sep 4;46(35):10039-45 Narayan N, Massimi P, Banks L. CDK phosphorylation of the discs large tumour suppressor controls its localisation and Abi-Char J, El-Haou S, Balse E, Neyroud N, Vranckx R, stability. J Cell Sci. 2009 Jan 1;122(Pt 1):65-74 Coulombe A, Hatem SN. The anchoring protein SAP97 retains Kv1.5 channels in the plasma membrane of cardiac myocytes. This article should be referenced as such: Am J Physiol Heart Circ Physiol. 2008 Apr;294(4):H1851-61 Massimi P, Banks L. DLG1 (discs, large homolog 1 Cao Z, Huett A, Kuballa P, Giallourakis C, Xavier RJ. DLG1 is (Drosophila)). Atlas Genet Cytogenet Oncol Haematol. 2010; an anchor for the E3 ligase MARCH2 at sites of cell-cell 14(4):368-376. contact. Cell Signal. 2008 Jan;20(1):73-82

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 376 Atlas of Genetics and Cytogenetics

in Oncology and Haematology OPEN ACCESS JOURNAL AT INIST-CNRS

Gene Section Review

EEF1A1 (eukaryotic translation elongation factor 1 alpha 1) Bruna Scaggiante, Giorgio Manzini Department of Life Sciences, University of Trieste, via Giorgieri, 1, 34147-Trieste, Italy (BS, GM)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/EEF1A1ID40407ch6q13.html DOI: 10.4267/2042/44731 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Transcription The main processed mRNA encompasses exons 1, 2, 3, Other names: CCS-3; CCS3; EEF-1; EEF1A; EF-1-; 4, 5, 6, 7, 8, this last can be in short or long form. In a alpha-1; EF-Tu; EF1A; FLJ25721; RAF-1EF; LENG7; few cases also exon 1' is retained. In a few cases exons MGC102687; MGC131894; MGC16224; PTI1; 1 (and 1') are substituted by the alternative exon from a eEF1A-1 putative upstream minor promoter, as described above. HGNC (Hugo): EEF1A1 Moreover, a quite high number of processed transcripts Location: 6q13 that, after exons 1 and 2, retain intron 2, which introduces a stop codon 22 residues downstream of Local order: Distal to LOC100129409, proximal to exon 2 are found (see for instance some of the many SLC17A5. ESTs: dbj-DC389722.1, dbj-DC341899.1, dbj- DNA/RNA DC414491.1). Pseudogene Description About 20 complete or approximately complete 8 exons, 7 introns (1st intron within 5'UTR), plus a rare intronless pseudogenes, likely generated by retro- optional exon within first intron as found in several transposition, a few of them exempt from frameshifs ESTs (e.g. emb-CR981691.1, dbj-DC388133.1, dbj- and with only a few missenses, are present throughout DC406334.1). the genome. Two of them harbour a few hundreds nt Presumably a second promoter, about 800 nt upstream long insert each, not related to introns of the expressed of the most common transcription start, provides an gene. alternative first exon about 320 nt long, as deduced All of them show a higher homology to EEF1A1 than from some ESTs at NCBI (e.g. dbj-DC316623.1, gb- to EEF1A2. Most of the pseudogenes find an BU173251.1, dbj-DC358918.1). orthologous counterpart within the chimpanzee Introns number 2, 3, 4, 6 are phase 0 (between codons), genome. Introns number 5, 7 are phase 1 (between 1 st and 2 nd EEF1AL3 (9q34) Note: highly homologous; EEF1AL4 base of codon). (7p15.3) Note: highly homologous but with 1 A validated C-G non-synonimous polymorphism has frameshift; EEF1AL5-LOC390924 (19q13.12) Note: been reported at 1st position of codon 382 (Arg-Gly), contains a 502 nt insert; EEF1AL6 (3q27.1); EEF1AL7 plus a few single-hit non-synonimous and some (4q24); EEF1AL8- synonymous within CDS. Several others within 3'UTR and introns (SNP source).

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 377 EEF1A1 (eukaryotic translation elongation factor 1 alpha 1) Scaggiante B, Manzini G

Box = exon (blue = 5'UTR, yellow = CDS, light blue = rare optional exon, red = 3'UTR, light red = extended 3'UTR to a downstream polyA signal) Line = intron.

Upper boxes, alternating colours: exons (coding part only). Lower boxes: protein domains.

LOC100132804 (7q35); EEF1AL9 (1p21.3); tRNA) to the A site of the ribosome as a ternary EEF1AL10-LOC644604 (2q12); EEF1AL11 (5p15.1) complex eEF1A1-GTP-aa-tRNA. In mammalian, it is Note: rather highly homologous; EEF1AL12- ubiquitously expressed with exception of skeletal LOC647167 (1q31.3); EEF1AL13-LOC100130211 muscle, heart and brain where during terminal (Xq21.2) Note: lacking about 300 initial codifying nt; differentiation eEF1A2 is produced (Knudsen et al., LOC124199 (16p12.1) Note: contains a 307 nt insert; 1993). LOC387845 (12p12.3); LOC389223 (4q28.3); Moonlighting functions: cytoskeletal remodel-ling, LOC401717 (12q12); LOC442709 (7q21.13) Note: protein folding and degradation, cell signal-ling harbours a 21 nt deletion; LOC645693 (15q21.2); modulation, control of cell growth, apoptosis and cell LOC645715 (3p22.1); LOC646612 (3q22.3) Note: cycle lacking about 120 initial codifying nt; LOC728672 1) eEF1A1 and cytoskeletal remodelling. The most (12p12.3); LOC100128082 (5p12). relevant non canonical function of eEF1A1 is the modulation of cytoskeleton organization. eEF1A has Protein activity on microtubule severing and bundling. It has a Note specific site to bind actin that is different from that for The major form of the protein is 462 residues long, the binding of aa-tRNA (Gross et al., 2005). eEF1A composed by three domains, as shown by the diagram, binding to F-actin is modulated by Rho/Rho-kinase that relates also the protein domains (lower bar) with pathway. Phospho-rylation by Rho kinase decreases the mRNA coding exons (upper bar). binding of eEF1A1 to F-actin and F-actin bundling. Myosin phosphatase acts in antagonist fashion on Description eEF1A1 to modulate actin cytoskeletal organization 462 residues, theoretical MW 50140.8 Da, theoretical (Izawa et al., 2000). isoelectric point 9.7 eEF1A1 is one of the alpha subunit 2) eEF1A1 and protein degradation and folding. forms of the elongation factor 1 complex, that interacts eEF1A controls translational fidelity by binding to with aminoacylated tRNA and delivers it to the A site uncorrectly folded proteins but not to correctly folded of the ribosome during the elongation phase of protein ones. The uncorrectly folded proteins are then directed synthesis. The other form is eEF1A2, encoded by a to degradation pathway (Hotokezaka et al., 2002). different gene, EEF1A2, located in chromosome 20. eEF1A plays a role in recognition and degradation of Expression co-translationally damaged and ubiquitylated proteins promoting their translocation to proteasome through EEF1A1 is constitutively expressed in all tissues, with interaction with proteasome subunit Rpt1 (Chuang and the exception of adult brain, heart and skeletal muscle, Madura, 2005). eEF1A exhibits chape-rone-like where EEF1A2 expression is found. activity by promoting renatura-tion of enzymes such as Localisation aminoacyl-tRNA synthetases, likely contributing to Mostly cytoplasmic, but also nuclear. maintain the efficiency of translational machinery (Lukash et al., 2004). Function 3) eEF1A1 and control of cell cycle, growth and Canonical function: aa-tRNA delivery to ribo-some in death. eEF1A as ribonucleoprotein complex, mRNA translation containing a non-coding RNA, binds to and mediates The eukaryotic elongation factor 1A (eEF1A1, activation of heat-shock transcription factor 1 (HSF1) formerly EF-1alpha or eEF1A) protein belongs to the to protect the cell from heat-shock (Shamovsky et al., G-protein superfamily, is one of most abundantly 2006). Induction of the non-constitutive eEF1A1 expressed protein in mammalian cells and participates expression in cardiomyocytes as response to lipotoxic to mRNA translation. It carries aminoacyl-tRNA (aa- ER-stress promotes cell death likely by activation of

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 378 EEF1A1 (eukaryotic translation elongation factor 1 alpha 1) Scaggiante B, Manzini G

eEF1A1-dependent cytoske-letal modifications the endothelial nitric oxide synthase (eNOS) to regulate triggering apoptosis (Borradaile et al., 2006). eEF1A1 post-translational eNOS mRNA stability. In the human interacts with the HDM2 gene product at a binding site endothelial cell line HUVEC, TNF-alpha-mediated for eEF1A1 overlaps with that for p53. In normal cells eNOS mRNA destabilization involves eEF1A1 to eEF1A1 could promote cell apoptosis by preventing reduce eNOS mRNA levels (Yan et al., 2008). p53 sequestra-tion by HDM2 (Frum et al., 2007). Homology Likely both eEF1A1 and eEF1A2 interacts with the zinc finger protein ZPR1 in response to mitogenic Highly homologous over the entire length to EEF1A2 stimuli, redistributing eEF1A1/2 and ZPR1 in the (92% identities); nucleus. This interaction is essential for normal cell Moderately homologous over all three domains, higher proliferation and growth. Thus the interaction for the first one, to : eEF1A1/2-ZPR1 is required for normal cell cycle HBS1L, a member of the GTP-binding protein family progression (Mishra et al., 2007). eEF1A1 is an expressed in erythroid progenitor cells (39% identities); interactor of Bood POZ containing gene type 2 (BPOZ- GSPT1, a GTP-binding protein involved in G1 to S 2) that promotes eEF1A1 ubiquitylation and phase transition (38% identities); degradation via 26S proteasome. BPOZ-2 inhibits GTP binding to eEF1A1 thus preventing translation. BOPZ- GSPT2, a GTP-binding protein involved in G1 to S 2 is transcriptionally activated by Phosphate and Tensin phase transition (37% identities); homologue deleted on chromosome 10 (PTEN). It has TUFM, Tu translation elongation factor, mitochon-drial been suggested that PTEN exerts growth inhibition (31% identities). effects in cells not only by antagonizing PI3K-Akt signalling path-way, but also inducing BPOZ-2 Implicated in expression to degrade eEF1A1. In this manner, in normal cells, the transition from growing to resting Head and neck cancers phases is mediated by BPOZ-2/eEF1A1 interaction, Note thus leading to prevention of translation and induction eEF1A1 overexpression is observed in cisplatin- of eEF1A1 degradation by 26S proteasome pathway resistant human head and neck cancer cell lines (Koiwai et al., 2008). eEF1A1 is implicated in a novel (Johnsson et al., 2000). cell cycle check-point to prevent tetraploidy in binucleated cells. In tetraploids, cell death, preventing Breast cancer aneuploidy malignancies, is mainly controlled in a Note caspase-independent man-ner by the down-regulation EEF1A1 was found to be upregulated in invasive breast of eEF1A1 levels. eEF1A1 mRNA accumulates in cancer cells derived from snap-frozen adenocarcinoma specialized P bodies to reduce the expression of the samples suggesting a role in mediating invasive activity proteins. The prominent signal in the eEF1A1 mRNA of cancer cells (Zhu et al., 2003). Treatment of the for its translational repression and degradation is in the breast human cancer cell line MCF-7 with the histone 5'-UTR. Exogenous expression of eEF1A1 inhibits cell deacetylase inhibitor sodium butyrate decreases death in tetraploids. Notably, exogenous expression of significantly in a dose-dependent manner the eEF1A1 eEF1A2 whose mRNA 5'-UTR differs from that of transcription levels. Thus, overexpression of eEF1A1 eEF1A1 inhibits cell death in tetraploids, thus contributes to breast cancer survival (Gonçalves et al., suggesting another mechanism by which eEF1A2 could 2005). promote tumour development (Kobayashi et al., 2009). Tongue squamous cell carcinoma 4) eEF1A1 and cell signalling modulation. Besides Note eEF1A2, in adult mouse neurons eEF1A1 is expressed too and it is able to regulate the recycle of M4 A suggestive down-regulation of EEF1A1 expression muscarinic acetylcholine receptors (mAChR). Thus, has been observed in human tongue squamous cell eEF1A1 plays a role in locomotor activity of neurons carcinoma with positive lympho-nodes and (McClatchy et al., 2006). eEF1A1 modulates the extracapsular spread. Thus EEF1A1 down-regulation activities of sphingosine kinases (SK1 and SK2). might be involved in the tumour cell progression Phosphorylated and non-phosphorylated eEF1A1 forms toward the metastasis (Zhou et al., 2006). interact with phosphoryla-ted and non phosphorylated Hepatocarcinoma SK1 and SK2 and this results in an increased enzymatic Note activity of both SK1 and SK2. In this respect, eEF1A1 overexpression in human hepatocarcinoma overexpression of eEF1A1 in quiescent cells has been cell lines correlates with an increase of proliferation suggested to play a role in oncogenesis by increasing rate and with the ability to escape apoptosis under SK1 and SK2 activities (Leclercq et al., 2008). eEF1A1 suboptimal growth conditions. In particular eEF1A1 is involved in the regulation of vascular function overexpression is higher in the more aggressive mediated by TNF-alpha. eEF1A1 binds to 3'-UTR of

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 379 EEF1A1 (eukaryotic translation elongation factor 1 alpha 1) Scaggiante B, Manzini G

phenotype cell line JHH6 with respect to the more known as elongation factor 1A-like 14. Initially differentiated HepG2 and HuH7 cells (Grassi et al., identified by differential RNA display screening of 2007). eEF1A1 regulates the half-life of osteopontin cDNA expression library in the human prostate cancer (OPN) mRNA. eEF1A1 is a trans-acting factor that cell line LNCaP, it has been proposed to act as a binds to 5'-UTR of OPN. This has strong implications dominant oncogene in human prostatic in the invasive process as demonstrated in adenocarcinoma. Its mRNA contains a 5'-UTR of 630 hepatocellular carcinoma cells, OPN being the major bp that is highly homologous to part of the 23S rRNA secreted phosphoprotein which is overexpressed by of Mycoplasma sp. fused to most of the CDS of tumour cells in advanced metastatic cancer. EEF1A1, resulting in the substitution of The higher expression of OPN in invasive cancer cells is due to the different localization of eEF1A1: in non its first 67 N-terminal residues with Met-Gln-Ser (Sun invasive Hep3B cells it is mainly bound to G actin et al., 1997; Mansilla et al., 2005). Ectopically forced whereas in invasive HepG2 type eEF1A1 and G actin expression of PTI-1 in a mouse cell line induces association is minimal. Thus eEF1A1 is an indirect tumours in nude mice and antisense PTI-1 molecule regulator of OPN by affecting the OPN mRNA stability can reverse malignant phenotype of the transformed through the interaction with G actin. Only F actin- cells (Su et al., 1998). Silencing of PTI-1 by specific bound eEF1A1 cannot interact with 5'-UTR of OPN RNAi not affecting eEF1A1 expression, in a human (Zhang et al., 2009). prostate cancer cell line leads to a reduction of cellular Testicular germ tumours growth, to the block of cell cycle in G1 phase and to the promotion of apoptosis (Yu et al., 2006). It has Note been hypothesized that PTI-1 could promote cell eEF1A1, as well as eEF1A2, is an interactor of the transformation by causing translational infidelity being human testis-specific Y-encoded (TSPY) gene. It has in competition with eEF1A1 (Gopalkrishnan et al., been demonstrated that the binding to TSPY leads to a 1999). PTI-1 mRNA is detected only in human cancer redistribution of the TSPY-eEF1A1/2 complex in the cells upon Mycoplasma infection. It remains under cell with a nuclear co-localization. A role of the TSPY- investigation whether PTI-1 can play a role in the eEF1A1/2 complex has been suggested in promoting natural history of human prostatic adenocarcinoma neoplastic transformation and in sustaining cancer cell upon Mycoplasma infection. The origin of the chimeric growth in human testicular germ tumours, prostate transcript of PTI-1 remains to be ascertained cancer, as well as in other somatic cancers (Kido et al., (Scaggiante et al., 2008). PTI-1 mRNA has been 2008). detected in multidrug-resistance colon cancer cell line Cervical cancer LoVoDX (Bertram et al., 1998). By using the detection Note of a unique PTI-1 region between the 5'UTR and the A variant form of eEF1A1 named cervical cancer CDS, PTI-1 mRNA has been found in the human suppressor 3 (CCS-3) lacking the 101 aminoacids at the pancreatic cancer cell line AsPC-1, in the human N-terminal region, has been identified as tumour gastric cancer TMK-1 cells, and in the hepatoma suppressor that is present in non-trans-formed human Alexander cells, but not in several other pancreatic, cell lines. Its ectopical expression in a cervical tumour gastric and hepatoma human cancer cell lines. cell line leads to cell growth inhibition and apoptosis. Interestingly, in AsPC-1 cells the down-regulation of CCS-3 seems to act as a co-transcriptional repressor by K-ras mRNA by antisense leads to a reduction of PTI-1 interacting with the transcriptional regulator PLZF level. PTI-1 mRNA was detected in three of five (Rho et al., 2006). surgical human specimens of pancreatic cancer (Ohnami et al., 1999). The mRNA of CCS-3 (GenBank Accession AF322220) appears to be a fusion product joining the reverse of the Cutaneous T-cell lymphoma end portion of the 3' UTR of another product Note (NM_005763, from gene AASS) with most of the In human sera derived from Cutaneous T-Cell normal sequence of EEF1A1 mRNA. Lymphoma (CTCL) patients one of the new tumour Various tumours including prostate antigens was a truncated version of eEF1A1 lacking 77 adenocarcinoma, colon aminoacids at N-terminal. adenocarcinoma, pancreatic cancer and Leukemia gastric cancer Note Note In the promyelocytic human leukemia cells the A variant form of eEF1A1 lacking 67 aminoacids at the differentiation agent All-Trans-Retinoic Acid (ATRA) N-terminal region has been proposed to promote cell induces down-regulation of eEF1A1 thus suggesting a transformation and to sustain tumour cells viability. It role in contributing to cancer survival in has been named prostate inducing gene-1 (PTI-1), also haematopoietic malignancies (Harris et al., 2004).

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 380 EEF1A1 (eukaryotic translation elongation factor 1 alpha 1) Scaggiante B, Manzini G

An isoform of eEF1A1 with a more basic isoelectric Ditzel HJ, Masaki Y, Nielsen H, Farnaes L, Burton DR. Cloning point was identified in human haematopoietic cancer and expression of a novel human antibody-antigen pair associated with Felty's syndrome. Proc Natl Acad Sci U S A. cell lines but not in normal lymphocytes raising the 2000 Aug 1;97(16):9234-9 possibility that post-translation modifications of Izawa T, Fukata Y, Kimura T, Iwamatsu A, Dohi K, Kaibuchi K. eEF1A1 could be involved in cancer development and Elongation factor-1 alpha is a novel substrate of rho-associated progression of haematopoietic tumours (Dapas et al., kinase. Biochem Biophys Res Commun. 2000 Nov 2003). 11;278(1):72-8 Felty's syndrome Johnsson A, Zeelenberg I, Min Y, Hilinski J, Berry C, Howell SB, Los G. Identification of genes differentially expressed in Note association with acquired cisplatin resistance. Br J Cancer. Characterized by rheumatoid arthritis, spleno-megaly 2000 Oct;83(8):1047-54 and neutropenia. eEF1A1 is found as autoantigen Hotokezaka Y, Tobben U, Hotokezaka H, Van Leyen K, Beatrix (Ditzel et al., 2000). B, Smith DH, Nakamura T, Wiedmann M. Interaction of the eukaryotic elongation factor 1A with newly synthesized Skeletal Muscle Trauma polypeptides. J Biol Chem. 2002 May 24;277(21):18545-51 Note Talapatra S, Wagner JD, Thompson CB. Elongation factor-1 In hypercatabolic traumatized patients eEF1A1 mRNA alpha is a selective regulator of growth factor withdrawal and level significantly rose in skeletal muscle as the result ER stress-induced apoptosis. Cell Death Differ. 2002 of injury. eEF1A1 expression correlated with Aug;9(8):856-61 overexpression of p66(ShcA) (Bosutti et al., 2007). Dapas B, Tell G, Scaloni A, Pines A, Ferrara L, Quadrifoglio F, Scaggiante B. Identification of different isoforms of eEF1A in Cell transformation the nuclear fraction of human T-lymphoblastic cancer cell line specifically binding to aptameric cytotoxic GT oligomers. Eur J Note Biochem. 2003 Aug;270(15):3251-62 De-regulation of eEF1A1 in rodent cells exposed to Zhu G, Reynolds L, Crnogorac-Jurcevic T, Gillett CE, Dublin chemical and physical carcinogens promotes cell EA, Marshall JF, Barnes D, D'Arrigo C, Van Trappen PO, transformation (Tatsuka et al., 1992). Over-expression Lemoine NR, Hart IR. Combination of microdissection and of eEF1A1 in a non-transformed murine pro-B cell line microarray analysis to identify gene expression changes confers selective resistance to apoptosis induced by between differentially located tumour cells in breast cancer. endoplasmic reticulum stress, thus providing long-term Oncogene. 2003 Jun 12;22(24):3742-8 viability (Talapatra et al., 2000). Harris MN, Ozpolat B, Abdi F, Gu S, Legler A, Mawuenyega KG, Tirado-Gomez M, Lopez-Berestein G, Chen X. Comparative proteomic analysis of all-trans-retinoic acid References treatment reveals systematic posttranscriptional control mechanisms in acute promyelocytic leukemia. Blood. 2004 Tatsuka M, Mitsui H, Wada M, Nagata A, Nojima H, Okayama Sep 1;104(5):1314-23 H. Elongation factor-1 alpha gene determines susceptibility to transformation. Nature. 1992 Sep 24;359(6393):333-6 Hartmann TB, Thiel D, Dummer R, Schadendorf D, Eichmüller S. SEREX identification of new tumour-associated antigens in Knudsen SM, Frydenberg J, Clark BF, Leffers H. Tissue- cutaneous T-cell lymphoma. Br J Dermatol. 2004 dependent variation in the expression of elongation factor-1 Feb;150(2):252-8 alpha isoforms: isolation and characterisation of a cDNA encoding a novel variant of human elongation-factor 1 alpha. Lukash TO, Turkivska HV, Negrutskii BS, El'skaya AV. Eur J Biochem. 1993 Aug 1;215(3):549-54 Chaperone-like activity of mammalian elongation factor eEF1A: renaturation of aminoacyl-tRNA synthetases. Int J Biochem Sun Y, Lin J, Katz AE, Fisher PB. Human prostatic carcinoma Cell Biol. 2004 Jul;36(7):1341-7 oncogene PTI-1 is expressed in human tumor cell lines and prostate carcinoma patient blood samples. Cancer Res. 1997 Chuang SM, Madura K. Saccharomyces cerevisiae Ub- Jan 1;57(1):18-23 conjugating enzyme Ubc4 binds the proteasome in the presence of translationally damaged proteins. Genetics. 2005 Bertram J, Palfner K, Hiddemann W, Kneba M. Dec;171(4):1477-84 Overexpression of ribosomal proteins L4 and L5 and the putative alternative elongation factor PTI-1 in the doxorubicin Gonçalves J, Malta-Vacas J, Louis M, Brault L, Bagrel D, resistant human colon cancer cell line LoVoDxR. Eur J Cancer. Monteiro C, Brito M. Modulation of translation factor's gene 1998 Apr;34(5):731-6 expression by histone deacetylase inhibitors in breast cancer cells. Clin Chem Lab Med. 2005;43(2):151-6 Su Z, Goldstein NI, Fisher PB. Antisense inhibition of the PTI-1 oncogene reverses cancer phenotypes. Proc Natl Acad Sci U Gross SR, Kinzy TG. Translation elongation factor 1A is S A. 1998 Feb 17;95(4):1764-9 essential for regulation of the actin cytoskeleton and cell morphology. Nat Struct Mol Biol. 2005 Sep;12(9):772-8 Gopalkrishnan RV, Su ZZ, Goldstein NI, Fisher PB. Translational infidelity and human cancer: role of the PTI-1 Mansilla F, Hansen LL, Jakobsen H, Kjeldgaard NO, Clark BF, oncogene. Int J Biochem Cell Biol. 1999 Jan;31(1):151-62 Knudsen CR. Deconstructing PTI-1: PTI-1 is a truncated, but not mutated, form of translation elongatin factor 1A1, eEF1A1. Ohnami S, Matsumoto N, Nakano M, Aoki K, Nagasaki K, Biochim Biophys Acta. 2005 Feb 14;1727(2):116-24 Sugimura T, Terada M, Yoshida T. Identification of genes showing differential expression in antisense K-ras-transduced Borradaile NM, Buhman KK, Listenberger LL, Magee CJ, pancreatic cancer cells with suppressed tumorigenicity. Cancer Morimoto ET, Ory DS, Schaffer JE. A critical role for eukaryotic Res. 1999 Nov 1;59(21):5565-71 elongation factor 1A-1 in lipotoxic cell death. Mol Biol Cell. 2006 Feb;17(2):770-8

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McClatchy DB, Fang G, Levey AI. Elongation factor 1A family SMN complexes. Proc Natl Acad Sci U S A. 2007 Aug regulates the recycling of the M4 muscarinic acetylcholine 28;104(35):13930-5 receptor. Neurochem Res. 2006 Jul;31(7):975-88 Kido T, Lau YF. The human Y-encoded testis-specific protein Rho SB, Park YG, Park K, Lee SH, Lee JH. A novel cervical interacts functionally with eukaryotic translation elongation cancer suppressor 3 (CCS-3) interacts with the BTB domain of factor eEF1A, a putative oncoprotein. Int J Cancer. 2008 Oct PLZF and inhibits the cell growth by inducing apoptosis. FEBS 1;123(7):1573-85 Lett. 2006 Jul 24;580(17):4073-80 Koiwai K, Maezawa S, Hayano T, Iitsuka M, Koiwai O. BPOZ-2 Shamovsky I, Ivannikov M, Kandel ES, Gershon D, Nudler E. directly binds to eEF1A1 to promote eEF1A1 ubiquitylation and RNA-mediated response to heat shock in mammalian cells. degradation and prevent translation. Genes Cells. 2008 Nature. 2006 Mar 23;440(7083):556-60 Jun;13(6):593-607 Yu L, Wu G, Wang L, Wang H, Zhang G. Transient reduction Leclercq TM, Moretti PA, Vadas MA, Pitson SM. Eukaryotic of PTI-1 expression by short interfering RNAs inhibits the elongation factor 1A interacts with sphingosine kinase and growth of human prostate cancer cell lines. Tohoku J Exp Med. directly enhances its catalytic activity. J Biol Chem. 2008 Apr 2006 Jun;209(2):141-8 11;283(15):9606-14 Zhou X, Temam S, Oh M, Pungpravat N, Huang BL, Mao L, Scaggiante B, Bonin S, Cristiano L, Siracusano S, Stanta G, Wong DT. Global expression-based classification of lymph Dapas B, Giansante C, Fiotti N, Grassi G. Prostate-tumor- node metastasis and extracapsular spread of oral tongue inducing gene-1 analysis in human prostate cancer cells and squamous cell carcinoma. Neoplasia. 2006 Nov;8(11):925-32 tissue in relation to Mycoplasma infection. Cancer Invest. 2008 Oct;26(8):800-8 Bosutti A, Scaggiante B, Grassi G, Guarnieri G, Biolo G. Overexpression of the elongation factor 1A1 relates to muscle Yan G, You B, Chen SP, Liao JK, Sun J. Tumor necrosis proteolysis and proapoptotic p66(ShcA) gene transcription in factor-alpha downregulates endothelial nitric oxide synthase hypercatabolic trauma patients. Metabolism. 2007 mRNA stability via translation elongation factor 1-alpha 1. Circ Dec;56(12):1629-34 Res. 2008 Sep 12;103(6):591-7 Frum R, Busby SA, Ramamoorthy M, Deb S, Shabanowitz J, Kobayashi Y, Yonehara S. Novel cell death by downregulation Hunt DF, Deb SP. HDM2-binding partners: interaction with of eEF1A1 expression in tetraploids. Cell Death Differ. 2009 translation elongation factor EF1alpha. J Proteome Res. 2007 Jan;16(1):139-50 Apr;6(4):1410-7 Zhang J, Guo H, Mi Z, Gao C, Bhattacharya S, Li J, Kuo PC. Grassi G, Scaggiante B, Farra R, Dapas B, Agostini F, Baiz D, EF1A1-actin interactions alter mRNA stability to determine Rosso N, Tiribelli C. The expression levels of the translational differential osteopontin expression in HepG2 and Hep3B cells. factors eEF1A 1/2 correlate with cell growth Exp Cell Res. 2009 Jan 15;315(2):304-12 but not apoptosis in hepatocellular carcinoma cell lines with This article should be referenced as such: different differentiation grade. Biochimie. 2007 Dec;89(12):1544-52 Scaggiante B, Manzini G. EEF1A1 (eukaryotic translation elongation factor 1 alpha 1). Atlas Genet Cytogenet Oncol Mishra AK, Gangwani L, Davis RJ, Lambright DG. Structural Haematol. 2010; 14(4):377-382. insights into the interaction of the evolutionarily conserved ZPR1 domain tandem with eukaryotic EF1A, receptors, and

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Gene Section Mini Review

FHL2 (four and a half LIM domains 2) Marie Lin, William Cheung Department of Chemistry, The University of Hong Kong, Hong Kong, PR. China (ML, WC)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/FHL2ID44092ch2q12.html DOI: 10.4267/2042/44732 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Transcription Four transcript variants of FHL2 genes have been Other names: AAG11; DRAL; FHL-2; SLIM3 reported in Entrez Gene (NCBI). These alternative HGNC (Hugo): FHL2 spliced transcripts are 1.55-1.91 kb in length, and differ Location: 2q12.2 in the 5'-UTR only. Local order; 91kb telomeric to transforming growth Pseudogene factor, beta receptor associated protein 1 (TGFBRAP1). No pseudogenes for FHL2 are known. DNA/RNA Protein Description Description Human FHL2 gene spans around 80kb of genomic The open reading frame encodes a 279 amino acid DNA on the chromosome 2q12-q14 in telomere-to- protein with an estimated molecular weight of 32.2kDa. centromere orientation. FHL2 promoter contains FHL2 protein constitutes four and a half putative transcription factor binding sites for SRF (serum response factor), NKX2-5, MEF-2, E2F and AP-1 (activator protein-1).

Chromosomal location of FHL2 gene (upper panel) and genomic organization of four FHL2 transcript variants.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 383 FHL2 (four and a half LIM domains 2) Lin M, Cheung W

N-terminal LIM domains. The four complete LIM domains extend from amino acid 40-92, 100-153, 162- Implicated in 217 and 220-275. Rhabdomyosarcoma Expression Note In human tissues, FHL2 expression is the most FHL2 expression is downregulated in rhabdomyo- abundant in adult heart and ovary, and of low level in sarcoma cells relative to normal myoblasts (Genini et brain, lung, liver, kidney and intestine. FHL2 is initially al., 1997). identified as a downregulated gene in human Hepatocellular carcinoma rhabdomyosarcoma cells. However, eleva-ted FHL2 expression is detected in other cancers, including Note hepatocellular carcinoma, glioblastoma, breast, In 8 of 10 human liver tumors samples, FHL2 mRNA prostate, ovarian, and gastrointestinal cancers. expression is higher than that in matched nontumor livers (Wei et al., 2003). In contrast, it is recently Localisation reported that FHL2 protein is down-regulated in liver Cytoplasm and nucleus. tumors, as compared with matched nontumor liver Function tissues. In addition, FHL2 inhibits hepatoma cell growth in vitro and in nude mice (Ding et al., 2009). At tissue level, FHL2 plays important roles in the development of cardiac circulatory system and Ovarian cancer placenta. It also induces osteoblast and myoblast Note differentiation. At cellular level, FHL2 participates in FHL2 protein expression is upregulated in epithelial various processes, including cell survival, adhesion, ovarian cancer, as compared with matched normal motility, transcription and signal trans-duction. At tissues (Gabriel et al., 2004). molecular level, the LIM domains of FHL2 are double zinc finger motifs that physically interact with partner Breast cancer proteins to modulate RNA splicing, DNA replication Note and repair. It also fun-ctions as a transcriptional co- FHL2 is overexpressed in human mammary carci-noma activator for androgen receptor, AP-1, CREB (cAMP samples, compared with normal breast tissues. FHL2 response element binding protein), CREM (cAMP induces the expression of cell cycle inhibitor response element modulator), BRCA1 (breast cancer p21Cip1/Waf1 in MDA-MB 231 breast cancer cells 1), WT-1 (wilms' tumor), and NF-kB (nuclear factor- (Martin et al., 2007). kB). Moreover, FHL2 is a transcriptional co-suppressor Prognosis for ERK2 (extracellular signal regulated kinase 2), SRF Patients with tumors expressing low amounts of FHL2 and FOXO1 (forkhead box O1). were characterized by a significantly better survival Homology compared to those with high intratumoral FHL2 expression (Gabriel et al., 2006). FHL2 belongs to the four-and-a-half-LIM-only protein family, which includes FHL1, FHL2, FHL3, FHL4 and Prostate cancer FHL5 (ACT). Human FHL2 amino acid sequence is Note 48.2% identical with FHL1, 53.4% with FHL3, 48.4% FHL2 expression is downregulated by 2- to 4-fold in with mouse FHL4, and 59.1% with FHL5. Orthologs of primary prostate cancer relatively to normal tissues for human FHL2 are found in macaque, mouse, rat, bovine, five pairs of samples (Kinoshita et al., 2005). Another dog, chicken, frog, zebrafish, amphioxis, drosophila study reports that FHL2 expression is increased in and C. elegans. prostate adenocarcinoma cells when compared with benign epithelial cells. It might be the subcellular Mutations localization of FHL2 that governs the progression of Somatic prostate cancer (Muller et al., 2002). Androgen-induced FHL2 expression is mediated by SRF (Heemers et al., G142A missense mutation, corresponding to Gly48Ser 2007). within the first LIM domain, is identified in Prognosis heterozygous state in a 49-years-old female dilated cardiomyopathy (DCM) patient. This mutation Nuclear, but not cytoplasmic expression of FHL2 abrogates the binding of FHL2 with titin/connectin, and significantly correlates with the recurrence of pros-tate in turn impairs the abnormal recruitment of metabolic cancer (Kahl el al., 2006). enzymes to cardiac sarcomere (Arimura et al., 2007).

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 384 FHL2 (four and a half LIM domains 2) Lin M, Cheung W

Gastrointestinal cancer prostate cancer compared with normal prostate. Int J Urol. 2005 Apr;12(4):390-7 Note Gabriel B, Fischer DC, Orlowska-Volk M, zur Hausen A, FHL2 expression is upregulated in gastric and Schüle R, Müller JM, Hasenburg A. Expression of the transcriptional coregulator FHL2 in human breast cancer: a colon cancer, compared with matched normal tissues. clinicopathologic study. J Soc Gynecol Investig. 2006 Suppression of FHL2 induces gastric and colon cell Jan;13(1):69-75 differentiation, and inhibits cell prolifera-tion and Johannessen M, Møller S, Hansen T, Moens U, Van Ghelue expression of oncogenes (survivin, cox-2, hTERT and M. The multifunctional roles of the four-and-a-half-LIM only c-jun) in vitro. Antisense FHL2 also inhibits protein FHL2. Cell Mol Life Sci. 2006 Feb;63(3):268-84 tumorigenesis of colon cancer cells in xenograft nude Kahl P, Gullotti L, Heukamp LC, Wolf S, Friedrichs N, mice model (Wang et al., 2007). Vorreuther R, Solleder G, Bastian PJ, Ellinger J, Metzger E, Schüle R, Buettner R. Androgen receptor coactivators lysine- Glioma specific histone demethylase 1 and four and a half LIM domain protein 2 predict risk of prostate cancer recurrence. Cancer Note Res. 2006 Dec 1;66(23):11341-7 The mRNA level of FHL2 is elevated in both low (3 of Arimura T, Hayashi T, Matsumoto Y, Shibata H, Hiroi S, 6) and high (11 of 13) grade glioma patient samples. Nakamura T, Inagaki N, Hinohara K, Takahashi M, Manatsu FHL2 induces glioblastoma cell prolifera-tion and SI, Sasaoka T, Izumi T, Bonne G, Schwartz K, Kimura A. migration in vitro, and promotes tumori-genesis in Structural analysis of four and half LIM protein-2 in dilated cardiomyopathy. Biochem Biophys Res Commun. 2007 May glioblastoma xenograft nude mice model. 25;357(1):162-7 Overexpression of FHL2 decreases mRNA levels of p53 and its downstream proapoptotic genes, and Heemers HV, Regan KM, Dehm SM, Tindall DJ. Androgen induction of the androgen receptor coactivator four and a half enhances promoter activities of AP-1, human LIM domain protein-2: evidence for a role for serum response telomerase reverse transcriptase and survivin genes (Li factor in prostate cancer. Cancer Res. 2007 Nov et al., 2008). 1;67(21):10592-9 Kleiber K, Strebhardt K, Martin BT. The biological relevance of References FHL2 in tumour cells and its role as a putative cancer target. Anticancer Res. 2007 Jan-Feb;27(1A):55-61 Genini M, Schwalbe P, Scholl FA, Remppis A, Mattei MG, Schäfer BW. Subtractive cloning and characterization of DRAL, Martin BT, Kleiber K, Wixler V, Raab M, Zimmer B, Kaufmann a novel LIM-domain protein down-regulated in M, Strebhardt K. FHL2 regulates cell cycle-dependent and rhabdomyosarcoma. DNA Cell Biol. 1997 Apr;16(4):433-42 doxorubicin-induced p21Cip1/Waf1 expression in breast cancer cells. Cell Cycle. 2007 Jul 15;6(14):1779-88 Chan KK, Tsui SK, Lee SM, Luk SC, Liew CC, Fung KP, Waye MM, Lee CY. Molecular cloning and characterization of FHL2, Wang J, Yang Y, Xia HH, Gu Q, Lin MC, Jiang B, Peng Y, Li a novel LIM domain protein preferentially expressed in human G, An X, Zhang Y, Zhuang Z, Zhang Z, Kung HF, Wong BC. heart. Gene. 1998 Apr 14;210(2):345-50 Suppression of FHL2 expression induces cell differentiation and inhibits gastric and colon carcinogenesis. Müller JM, Metzger E, Greschik H, Bosserhoff AK, Mercep L, Gastroenterology. 2007 Mar;132(3):1066-76 Buettner R, Schüle R. The transcriptional coactivator FHL2 transmits Rho signals from the cell membrane into the nucleus. Li M, Wang J, Ng SS, Chan CY, Chen AC, Xia HP, Yew DT, EMBO J. 2002 Feb 15;21(4):736-48 Wong BC, Chen Z, Kung HF, Lin MC. The four-and-a-half-LIM protein 2 (FHL2) is overexpressed in gliomas and associated Wei Y, Renard CA, Labalette C, Wu Y, Lévy L, Neuveut C, with oncogenic activities. Glia. 2008 Sep;56(12):1328-38 Prieur X, Flajolet M, Prigent S, Buendia MA. Identification of the LIM protein FHL2 as a coactivator of beta-catenin. J Biol Ding L, Wang Z, Yan J, Yang X, Liu A, Qiu W, Zhu J, Han J, Chem. 2003 Feb 14;278(7):5188-94 Zhang H, Lin J, Cheng L, Qin X, Niu C, Yuan B, Wang X, Zhu C, Zhou Y, Li J, Song H, Huang C, Ye Q. Human four-and-a- Gabriel B, Mildenberger S, Weisser CW, Metzger E, Gitsch G, half LIM family members suppress tumor cell growth through a Schüle R, Müller JM. Focal adhesion kinase interacts with the TGF-beta-like signaling pathway. J Clin Invest. 2009 transcriptional coactivator FHL2 and both are overexpressed in Feb;119(2):349-61 epithelial ovarian cancer. Anticancer Res. 2004 Mar- Apr;24(2B):921-7 This article should be referenced as such: Kinoshita M, Nakagawa T, Shimizu A, Katsuoka Y. Differently Lin M, Cheung W. FHL2 (four and a half LIM domains 2). Atlas regulated androgen receptor transcriptional complex in Genet Cytogenet Oncol Haematol. 2010; 14(4):383-385.

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Gene Section Mini Review

ING2 (inhibitor of growth family, member 2) Susanne Jennek, Aria Baniahmad Institute of Human Genetics and Anthropology, Jena University Hospital, Kollegiengasse 10, 07743 Jena, Germany (SJ, AB)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/ING2ID40975ch4q35.html DOI: 10.4267/2042/44733 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Protein Other names: ING1L; ING1Lp; p32; p33ING2 Description HGNC (Hugo): ING2 p33ING2a: 280 amino acids; 33kDa protein; harbor, Location: 4q35.1 listed from the N-terminal to the C-terminal region, a leucine-zipper-like-region (LZL), a novel conserved DNA/RNA region (NCR), a nuclear localization signal (NLS), a plant homeo domain (PHD) finger motif and a poly Description basic region (PBR); p28ING2b: 240 aa; 28 kDa The two isoforms share exon 2 but have different exon protein; lacks leucine-zipper-like-domain (LZL) is 1. The exon 1a of ING2a encodes 58 amino acids. Exon distinct to the N-termi-nal part to ING2a (Unoki et al., 1b of ING2b encodes 18 amino acids. There is no 2008). significant homology between the N-terminal part of Expression ING2b and the N-terminal regions of ING1 isoforms ING2 is widely expressed in normal tissues (Shimada (Unoki et al., 2008). et al., 1998). Transcription Localisation The promoter region of ING2a possesses two p53 ING2 is predominantly localized in the nucleus to binding sites in contrast to the promoter of ING2b. chromatin and the nuclear matrix (Gozani et al., 2003). Binding of p53 to these sites suppresses the ING2a Through reduced levels of phosphoinositide PtdIns5P, expression. The promoter region of ING2b harbors a ING2 might be released from chromatin and HSF1 and HSF2 binding site, a c-Rel, a SP1 and five translocates partially to the cytoplasm (Gozani et al., MZF1 binding sites (Unoki et al., 2008). 2003).

Gene structure of ING2a and ING2b (modified according to Unoki et al., 2008).

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 386 ING2 (inhibitor of growth family, member 2) Jennek S, Baniahmad A

Function Hepatocellular carcinoma (HCC) The tumor suppressor protein ING2 has been described Oncogenesis to regulate cell cycle, cellular senescence and gene ING2 transcription and post-transcription level is regulation including chromatin level. In response to downregulated in the majority of HCC tumors DNA damage ING2 enhances the acetylation of p53, compared with non-tumors liver tissue. Further-rmore, which negatively regulates cell proliferation ING2 expression level is reduced in 44 of 84 (52.4%) (Nagashima et al., 2001). In addition the level of ING2 HCC cases and the ING2 expression level correlated expression directly regulates the onset of replicative with tumor size, histopathologic classifi-cation and senescence through the induc-tion of p300-dependent serum AFP (Zhang et al., 2008). It is also shown that acetylation of p53 (Pedeux et al., 2005). ING2 also HCC patients with reduced ING2 expression have a induces the global histone H4 acetylation and significantly increased risk exhibiting a shorter survival chromatin relaxation and thereby enhances the time. In conclusion, ING2 may be involved in the nucleotide excision repair (Wang et al., 2006). progression of HCC (Zhang et al., 2008). Furthermore, ING2 is also a part of two related Head and neck squamous cell mSin3/HDAC1/HDAC2 corep-ressor complexes (Doyon et al., 2006). Further, chromatin association of carcinoma (HNSCC) ING2 is linked to ING2 ability to bind to trimethylated Oncogenesis K4 of histone 3 (H3K4me3) via its plant homeodomain There is a loss of heterozygosity (LOH) in the region (PHD) region (Shi et al., 2006). p33ING2 is also 4q32 in the long arm of the chromosome 4 in 20% of associated with histone methyltranferase (HMT-) the cases (Borkovsky et. al., 2008). This region activity in vitro and in vivo, methylating specifi-cally includes ING2 and SAP30 genes that are parts of the histone H3 and histone H1 (Goeman et al., 2008). In two related mSin3/ HDAC1/2 corepres-sor complexes. addition, ING2, as a transcriptional rep-ressor, directly LOH on region 4q35.1 was detected in 30 (54.6%) out interacts with the corepressor Alien and enhances the of 55 informative cases (Borkovsky et. al., 2008). High Alien-mediated gene silencing (Fegers et al., 2007). LOH frequency is associated with advanced tumor Furthermore, ING2 interacts with the Smad-interaction stages; therefore, ING2 LOH is likely to be a late event transcriptional modulator SnoN mediating TGF-beta- in HNSCC. induced Smad-depen-dent transcription and cellular responses (Sarker et al., 2008). The activity of ING2, as Lung cancer a nuclear phosphatidylinositol receptor can be Oncogenesis modulated by phosphoinositides (Gozani et al., 2003). Although, there are no ING2 mutations identified in 30 human lung cancer cell lines and 31 primary lung Homology cancer tumors. The ING2 mRNA expression is reduced The PHD-finger motif is highly-conserved among all in 6 out of 7 lung cancer cell lines (Okano et al., 2006). ING genes. There are five human ING genes (ING1, ING2, ING3, ING4, ING5) which encode multiple Cutaneous melanomas isoforms via splicing. So far known ING2 gene Oncogenesis encodes two isoforms (ING2a: 33kDa; ING2b: The nuclear expression level of ING2 is significantly 28kDA). reduced in human melanomas compared to dysplastic nevi. It is suggested that reduced ING2 expression may Mutations be involved in the initiation of melanoma (Lu et al., 2006; Ythier et al., 2008). Note So far natural occurring point mutations of ING2 in References association with cancer were not yet described. However, loss of heterozygosity (LOH) and aberrant Shimada Y, Saito A, Suzuki M, Takahashi E, Horie M. Cloning of a novel gene (ING1L) homologous to ING1, a candidate ING2-mRNA levels were associated with cancer. tumor suppressor. Cytogenet Cell Genet. 1998;83(3-4):232-5 Nagashima M, Shiseki M, Miura K, Hagiwara K, Linke SP, Implicated in Pedeux R, Wang XW, Yokota J, Riabowol K, Harris CC. DNA damage-inducible gene p33ING2 negatively regulates cell Colon cancer proliferation through acetylation of p53. Proc Natl Acad Sci U S Oncogenesis A. 2001 Aug 14;98(17):9671-6 ING2 expression level in human colon tumors is Gozani O, Karuman P, Jones DR, Ivanov D, Cha J, Lugovskoy significantly higher than in normal colon tissue. In AA, Baird CL, Zhu H, Field SJ, Lessnick SL, Villasenor J, conclusion, ING2 might be involved in colon cancers Mehrotra B, Chen J, Rao VR, Brugge JS, Ferguson CG, Payrastre B, Myszka DG, Cantley LC, Wagner G, Divecha N, (Shimada et al., 1998). Prestwich GD, Yuan J. The PHD

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 387 ING2 (inhibitor of growth family, member 2) Jennek S, Baniahmad A

finger of the chromatin-associated protein ING2 functions as a Fegers I, Kob R, Eckey M, Schmidt O, Goeman F, nuclear phosphoinositide receptor. Cell. 2003 Jul 11;114(1):99- Papaioannou M, Escher N, von Eggeling F, Melle C, 111 Baniahmad A. The tumor suppressors p33ING1 and p33ING2 interact with alien in vivo and enhance alien-mediated gene Campos EI, Chin MY, Kuo WH, Li G. Biological functions of the silencing. J Proteome Res. 2007 Nov;6(11):4182-8 ING family tumor suppressors. Cell Mol Life Sci. 2004 Oct;61(19-20):2597-613 Soliman MA, Riabowol K. After a decade of study-ING, a PHD for a versatile family of proteins. Trends Biochem Sci. 2007 Pedeux R, Sengupta S, Shen JC, Demidov ON, Saito S, Onogi Nov;32(11):509-19 H, Kumamoto K, Wincovitch S, Garfield SH, McMenamin M, Nagashima M, Grossman SR, Appella E, Harris CC. ING2 Goeman F, Otto K, Kyrylenko S, Schmidt O, Baniahmad A. regulates the onset of replicative senescence by induction of ING2 recruits histone methyltransferase activity with p300-dependent p53 acetylation. Mol Cell Biol. 2005 methylation site specificity distinct from histone H3 lysines 4 Aug;25(15):6639-48 and 9. Biochim Biophys Acta. 2008 Oct;1783(10):1673-80 Doyon Y, Cayrou C, Ullah M, Landry AJ, Côté V, Selleck W, Sarker KP, Kataoka H, Chan A, Netherton SJ, Pot I, Huynh Lane WS, Tan S, Yang XJ, Côté J. ING tumor suppressor MA, Feng X, Bonni A, Riabowol K, Bonni S. ING2 as a novel proteins are critical regulators of chromatin acetylation required mediator of transforming growth factor-beta-dependent for genome expression and perpetuation. Mol Cell. 2006 Jan responses in epithelial cells. J Biol Chem. 2008 May 6;21(1):51-64 9;283(19):13269-79 Lu F, Dai DL, Martinka M, Ho V, Li G. Nuclear ING2 Unoki M, Kumamoto K, Robles AI, Shen JC, Zheng ZM, Harris expression is reduced in human cutaneous melanomas. Br J CC. A novel ING2 isoform, ING2b, synergizes with ING2a to Cancer. 2006 Jul 3;95(1):80-6 prevent cell cycle arrest and apoptosis. FEBS Lett. 2008 Nov 26;582(28):3868-74 Okano T, Gemma A, Hosoya Y, Hosomi Y, Nara M, Kokubo Y, Yoshimura A, Shibuya M, Nagashima M, Harris CC, Kudoh S. Ythier D, Larrieu D, Brambilla C, Brambilla E, Pedeux R. The Alterations in novel candidate tumor suppressor genes, ING1 new tumor suppressor genes ING: genomic structure and and ING2 in human lung cancer. Oncol Rep. 2006 status in cancer. Int J Cancer. 2008 Oct 1;123(7):1483-90 Mar;15(3):545-9 Zhang HK, Pan K, Wang H, Weng DS, Song HF, Zhou J, Perez-Ordoñez B, Beauchemin M, Jordan RC. Molecular Huang W, Li JJ, Chen MS, Xia JC. Decreased expression of biology of squamous cell carcinoma of the head and neck. J ING2 gene and its clinicopathological significance in Clin Pathol. 2006 May;59(5):445-53 hepatocellular carcinoma. Cancer Lett. 2008 Mar 18;261(2):183-92 Shi X, Hong T, Walter KL, Ewalt M, Michishita E, Hung T, Carney D, Peña P, Lan F, Kaadige MR, Lacoste N, Cayrou C, Borkosky SS, Gunduz M, Nagatsuka H, Beder LB, Gunduz E, Davrazou F, Saha A, Cairns BR, Ayer DE, Kutateladze TG, Shi Ali MA, Rodriguez AP, Cilek MZ, Tominaga S, Yamanaka N, Y, Côté J, Chua KF, Gozani O. ING2 PHD domain links Shimizu K, Nagai N. Frequent deletion of ING2 locus at 4q35.1 histone H3 lysine 4 methylation to active gene repression. associates with advanced tumor stage in head and neck Nature. 2006 Jul 6;442(7098):96-9 squamous cell carcinoma. J Cancer Res Clin Oncol. 2009 May;135(5):703-13 Wang J, Chin MY, Li G. The novel tumor suppressor p33ING2 enhances nucleotide excision repair via inducement of histone This article should be referenced as such: H4 acetylation and chromatin relaxation. Cancer Res. 2006 Feb 15;66(4):1906-11 Jennek S, Baniahmad A. ING2 (inhibitor of growth family, member 2). Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4):386-388.

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Gene Section Review

KLK7 (kallikrein-related peptidase 7) Ying Dong, John Lai, Judith A Clements Hormone Dependent Cancer Program, Institute of Health and Biomedical Innovation (IHBI), Queensland University of Technology (QUT), Brisbane, Australia (YD, JL, JAC)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/KLK7ID41087ch19q13.html DOI: 10.4267/2042/44734 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

and ovarian cells, suggesting the expression of tissue Identity specific KLK7 transcripts. Other names: PRSS6; SCCE; hK7; hSCCE Pseudogene HGNC (Hugo): KLK7 Not identified. Location: 19q13.33 Local order: Telomere to centromere. Protein DNA/RNA Description Full-length KLK7 (253 amino acids) has a secretion Description signal (pre-) peptide (22 amino acids), followed by an The gene encompasses 6.509 kb of gDNA. activation (pro-) peptide (7 amino acids) and the mature chain (224 amino acids) with 1 potential N-linked Transcription glycosylation site. The catalytic triad of His 70 , Asp 112 Five variant mRNA transcripts have been identi-fied. and Ser 205 (relative to ATG1) is conserved and is These include transcripts using different 5' untranslated essential for proteolytic activity. After synthesis as a regions (UTRs) including exon 1 deletions, and KLK7 full-length protein, the signal peptide is then transcripts using different 3'UTR regions. Using rapid cleaved and pro-KLK7 (zymogen) is subsequently amplification of cDNA ends (RACE) different KLK7 secreted from the cell. On activation, the propeptide is 5'UTR sequences were identified from RNA extracted removed and the zymogen becomes the mature active from pancreas, skin enzyme.

Genomic and protein structure of the KLK7 gene. The KLK7 gene is classically comprised of 5 coding exons (red boxes, classic numerals) and 4 intervening introns with a conserved intron phase pattern (I, II, I, 0). A non-coding exon and untranslated regions are shown in yellow. Also shown is the classical transcription initiation site (TIS) and corresponding translation start site (ATG1). An exon 1 deleted transcript has also been identified which would potentially result in the use of an alternative translation start site (ATG2). The numbering for the amino acid residues of the catalytic triad (His 70 , Asp 112 , Ser 205 ) are relative to the full-length protein starting from ATG1.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 389 KLK7 (kallikrein-related peptidase 7) Dong Y, et al.

KLK7 can complex with antileukoprotease (secretory human body fluid, such as, seminal plasma, breast leukocyte protease inhibitor), elafin, Lympho-epithelial milk, ovarian cancer ascites, salivary and Kazal type inhibitor (LEKTI) fragments, and a member cervicovaginal fluid. of a2-macroglobulin (a2M) protease inhibitor family, In normal skin, KLK7 is expressed in late epidermal a2-macroglobulin-like 1 (a2ML1). differentiation and found at all sites of epithelial X-ray structures of recombinant full-length KLK7 from cornification. Consequently, KLK7 is used as a marker E. coli and insect cells have been solved, from which for terminal epidermal differentia-tion. In normal the most distinguishing features of KLK7 are the short epidermis, KLK7 was detected in a population of 70-80 loop and the unique S1 pocket, which prefers P1 dendritic cells and in high suprabasal keratinocytes. Tyr residues. KLK7 displays a unique chymotrypsin- KLK7 was also found in reconstruct-ted human like specificity for Tyr, which is preferred at P2. In epidermis and its expression was suppressed by retinoic addition, KLK7 exhibits large positively charged acid. An increased expres-sion of KLK7 was found in surface patches, representing putative exosites for suprabasal cells in orthokeratotic and parakeratotic prime side substrate recognition. areas of the lesions of oral lichen planus (an Similar to several other KLKs and based on the binding inflammatory disease) and benign oral keratosis (a non- inflammatory disease). of metal to histidine such as His 99 , the KLK7 activity is inhibited by Zn ++ and Cu ++ at low micromolar High KLK7 protein levels have been detected in the concentrations. KLK7 induced degradation of tissues of lung, breast, ovarian and squamous cervical corneodesmosin and desmocollin 1 with similar cancers, oral squamous cell carcinoma and cervical efficiency in acidic (pH 5.6) and neutral (pH 7.2) adenocarcinoma tissues from patients. However, KLK7 conditions. KLK7 activity is modulated by water is down regulated in cancerous prostate tissues content in stratum corneum as KLK7 activity increased compared to normal prostate tissues. KLK7, along with significantly in an environment of high relative KLK6 and KLK10, is decreased in cerebrospinal fluid humidity. KLK7 also demonstrated a tolerance to water of frontotemporal dementia patients. restriction suggesting that it may be adapted to function Localisation in the water-restricted stratum corneum. Thus, relative humidity modulates desquamation by its effect upon Full-length KLK7 is localised intracellularly in the KLK7 activity, possibly other desquamatory hydrolases cytoplasm prior to secretion. KLK7 protein is co- and adapted KLK7 function in water-deplete skin. localised with KLK5 in skin and acinar cells of the pancreas by immunohistochemical staining. An N-terminal truncated KLK7 isoform (181 amino acids) initiating from the putative ATG2 would not The putative N-terminal truncated KLK7-181 isoform have the pre-pro-region and 43 amino acids from the is potentially not secreted as it does not have a signal N-terminus of full-length KLK7. The histidine which is peptide, and cellular localisation remains to be part of the catalytic triad is also omitted which would determined. result in a proteolytic inactive protein. The presence of Function this isoform has not yet been confirmed in human tissues or biological fluids. To date, the major biological functions of KLK7 are associated with the skin and related epithelial tissues, Expression such as hair follicles, oral mucosa and glandular Full-length KLK7 protein was originally purified in lobules. KLK7 is involved in keratiniza-tion, stratum human skin and named stratum corneum chymotryptic corneum formation, and turnover/ desquamation of the enzyme (SCCE, hSCCE). KLK7 cDNA was originally skin through the degradation of cell adhesion isolated from a keratinocyte derived library and glycoproteins, such as corneodes-mosin, desmocollin 1 designated PRSS6. Although Northern blot analyses and plakoglobin. KLK7 has also been shown to cleave have shown that KLK7 mRNA is predominantly insulin B chain, degrade fibronectin, fibrinogen and localised to skin and pancreas, more sensitive RT-PCR interleukin 1beta (IL-1b), as well as activate pro-IL-1b. experiments have shown that brain, kidney, ovary, KLK7 and KLK5 can control activation of the human bone, breast, endometrium, spinal cord, lung, prostate cathelicidin precursor protein, hCAP18, implying their tissue and salivary tissue express KLK7 mRNA at low ability to control innate immune defence. to modest levels. High KLK7 mRNA has been detected An in vitro study showed that UVB radiation can in malignancies of ovary, breast, lung and brain. increase KLK7 and KLK5 expression at both mRNA KLK7 protein has been detected by ELISA in a wide and protein levels in keratinocyte (HaCat) cells. In the range of tissues at low (adrenal, bladder, cervix, epidermis, the major inhibitor of KLK7, fallopian tube, kidney, lung, lymph node, muscle, antileukoprotease (secretory leukocyte protease ovary, salivary gland, small intestine, spinal cord, inhibitor) is produced by keratinocytes and can inhibit spleen, thyroid gland, tonsil, trachea and vagina) to detachment of corneocytes from human plantar callus high (oesophagus, heart, liver and skin) levels. Modest in vitro, while a weaker KLK7 inhibitor, elafin (skin- levels of KLK7 protein have also been detected in derived antileukoprotease), can reduce the shedding of

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 390 KLK7 (kallikrein-related peptidase 7) Dong Y, et al.

corneocytes. Established epidermal mouse models 1 deleted short KLK7 transcripts were detected in overexpressing KLK7 have been shown to develop serous EOC cells, while none or only KLK7 short chronic itchy dermatitis. Further characterisation of transcript was found in normal ovarian epithelial cells. these models also revealed epidermal In addition, a coordinated expression pattern and co- hyperproliferation, decreased skin barrier function, and localisation of KLK7 and KLK5 were found in serous decreased expression of MHC II antigen in EOC cells suggesting a proteolytic cascade between keratinocytes. These data provide an in vivo them. Co-overexpression of KLK4, KLK5, KLK6 and pathophysiological foundation that KLK7 plays an KLK7 in ovarian cancer cells (OV-MZ-6) led to important role in skin, such as listed those below. increased invasion in vitro and resulted in increased KLK cascade activation systems have been des-cribed. tumour burden in nude mice. A coordinated expression KLK7 is activated by KLK5 and KLK12. KLK7 of KLK7 and protease inhibitor antileukoprotease was activates other members of the kallikrein-related also found in EOC cells. Of interest, the 110-139 amino acid region of the KLK7 protein incorporates multiple peptidase family including KLK1, KLK2, prostate + + specific antigen (PSA/KLK3) and KLK9. CD8 CTL and CD4 helper T cell epitopes, and represents an attractive target antigen for The function of the putative N-terminal truncated immunotherapy of ovarian cancer. KLK7 remains to be established. Prognosis Homology EOC patients with KLK7 mRNA or protein expression At the protein level, KLK7 shares 28% (KLK12), 33% in their tumours had a significantly shorter disease-free (KLK9), 36% (KLK10, 11), 37% (KLK1, KLK3/PSA), survival time than those with KLK7 negative tumours. 38% (KLK2, KLK5, KLK6, KLK13), 40% (KLK8), KLK7 is an independent unfavourable predictor of 41% (KLK14), 43% (KLK4) and 42.6% (KLK15) disease-free and overall survival for patients with low sequence homology with other members of the grade cancers. KLK7 has been shown to increase kallikrein-related peptidase family. specificity for diagnosis and prognosis of EOC in conjunction with other biomarkers, such as CA125, Mutations HE4 and B7-H4. Germinal Breast cancer An AACC insertion in the 3'UTR of the KLK7 gene Disease has been found, which altered the common allele KLK7 gene expression was significantly lower in AACC to the rare allele AACCAACC. This insertion tumour tissues from early stage (I/II) breast cancer was found to be associated with atopic dermatitis. patients and tumour cells with progesterone receptors. Prognosis Implicated in Two groups have reported conflicting data regarding Endocrine related cancers the prognoses for breast cancer patients and KLK7 expressing tumours. One study found that breast cancer Disease patients with KLK7 positive tumours have relatively It has been postulated that KLK7 plays a role in shorter disease-free survival and overall survival than endocrine related cancers given its (a) dysregulated patients with KLK7 negative tumours. However, expression in cancerous tissues compared to normal another study reported that breast cancer patients with tissues, (b) regulation by hormones, such as, oestradiol, KLK7 expressing tumours have favourable outcomes progestins and glucocorticoids and (c) potential roles in compared to those with KLK7 negative tumours. degradation of cell-cell adhesion proteins, extracellular matrix (ECM) proteins and activation of other Cervical cancer proteases and growth factors. Disease Epithelial ovarian carcinoma (EOC) In a study of 18 cervical cancer cell lines (10 primary and 8 established cell lines) and 8 normal cervical Disease keratinocyte cell lines, KLK7 mRNA expression was Kallikrein 7 is highly expressed in serous EOC at both detected in the cancer cells (5/10 primary and 4/8 the mRNA and protein levels, and high KLK7 mRNA established lines) but not in any of the normal cervical expression is associated with poorly differentiated, late keratinocytes. Interestingly, all five patients, who clinical stage ovarian carcinomas and the volume of harbour KLK7 positive tumours that were used to residual tumour after surgery. Upregulated KLK7 establish the primary cell lines, were found to have protein was detected in EOC patient sera and tumour metastatic involvement of the pelvic tumour draining cytosols using ELISA, and in EOC tissue sections lymph nodes. In the same study, tumour restricted using immunohistochemis-try and a quantitative expression of KLK7 was confirmed by automated in situ immuno-fluorescence-based protein immunohistochemistry staining in 4 of the 5 primary analysis. At the mRNA level, both KLK7 and its exon

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squamous cervical tumours, and 1 of the 4 primary KLK7 expression from a survival analysis study of 73 adenocarcinomas, but none of the normal cervical patients with intracranial tumours. Overexpression of epithelial cells. Another immuno-histochemical study KLK7 protein by cultivated brain tumour cells showed a significantly higher expression of KLK7 in significantly enhanced the invasive potential in a cervical adenocarcinomas compared to normal Matrigel invasion assay. endocervical glands. Colon cancer Pancreatic cancer Disease Disease One study using a semi-quantitative RT-PCR method KLK7 is expressed in normal pancreas at mRNA and showed that the KLK7 gene is up-regulated in colon protein levels, and KLK7 protein is localised in acinar cancer and its expression predicts poor prognosis for cells of the pancreas by immunohistoche-mical colon cancer patients. staining. KLK7 is overexpressed in pan-creatic adenocarcinomas at both the mRNA and protein levels. Skin disorders KLK7 expression was also observed in neoplastic cells Note of all tumours examined using immunohistochemistry A majority of studies have concentrated on the with moderate-to-intense staining in 16 of the 23 concomitant functions of KLK7 and KLK5 in normal tumours examined. Only 2 of the 13 nonmalignant human skin and a number of skin disorders given its (a) tissue specimens displayed moderate KLK7 staining, high expression in pathological conditions compared to whereas the remaining specimens showed weak normal skin samples, (b) cleavage/degradation of immunoreactivity. In pancreatic cancer cells, KLK7 intercellular adhesive glycolproteins and (c) potential was shown to i) cleave desmoglein 2, ii) cleave E- of activation and degradation of cytokines, such as cadherin and the ECM protein, fibronectin, iii) enhance interleukin 1beta (IL-1b). urokinase-type plasminogen activator receptor shedding, and iv) reduce cell aggregation and adhesion Netherton syndrome (NS) to vitronectin to promote pancreatic cancer invasion. Disease Oral squamous cell carcinoma (OSCC) NS is a rare but severe inherited disorder that presents Disease the three following characteristics with varying degrees cDNA microarray analysis revealed that KLK5, KLK7, of severity of their symptoms. 1) Ichthyosiform KLK8 and KLK10 were upregulated in tumour samples erythroderma - inflamed, red, scaly skin and versus normal controls. RT-qPCR analysis confirmed trichorrhexis invaginata ("bamboo hair"). 2) Short, that KLK7 mRNA was most differentially regulated brittle, lustreless hair and atopic diathesis. 3) with a 5.3-fold increase, while 2.8-, 4.0- and 3.5-fold Predisposition to allergy problems. increases were observed for KLK5, KLK8, and NS patients have mutations in the serine protease KLK10, respectively. Immunohistochemical analysis inhibitor Kazal-type 5 (SPINK5) gene, encoding the demonstrated strong reactivity for all 4 KLK proteins in protease inhibitor LEKTI (lympho-epithelial Kazal- both orthotopic murine tumours and human OSCC type related inhibitor). Early studies using mouse tissues. models revealed that SPINK5-deficient mice mimic NS through degradation of desmoglein 1 by epidermal Lung cancer protease. The pathophysiological pro-cesses in the skin Disease and epithelial related tissues of NS patients result from KLK7 mRNA levels are decreased in adenocarci-noma the lack of functional LEKTI protease inhibitor and compared to matched nonmalignant lung tissue. consequently the over-degradation of Similarly, sera of patients with non-small cell lung corneodesmosomal cadherins by KLK7, KLK5 and cancer (NSCLC) have lower protein levels of KLK7, KLK14. KLK5, KLK8, KLK10 and KLK12 than those from The SPINK5 gene is localised chromosome 5. SPINK5 normal subjects. However, a study has reported intense mutations introduce premature termination codons in cytoplasmic staining for KLK7, KLK5, KLK6 and LEKTI transcripts and lead to the production of KLK8 in 40-90% of squamous cell carcinomas, small truncated LEKTI forms that lack several inhibitory cell carcinomas and carci-noid tumours while 20% of domains. NS is an autosomal recessive condition. tumour cells had intense nuclear staining for KLK7, KLK5 and KLK8. Atopic dermatitis (AD) Brain tumours Disease AD is a chronic inflammatory and allergic skin Disease disorder. Multifactorial studies have suggested that RT-qPCR analysis showed that KLK7 mRNA both genetic and environmental factors contribute to expression in intracranial tumours was associated with AD development. A study comprising 103 AD patients shorter overall survival than those tumours with no

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 392 KLK7 (kallikrein-related peptidase 7) Dong Y, et al.

and 261 matched controls revealed a significant Skytt A, Strömqvist M, Egelrud T. Primary substrate specificity association between the rare AACCAACC allele in the of recombinant human stratum corneum chymotryptic enzyme. Biochem Biophys Res Commun. 1995 Jun 15;211(2):586-9 3'UTR of KLK7 with AD. However, another group found that the AACCAACC allele was not associated Sondell B, Thornell LE, Egelrud T. Evidence that stratum corneum chymotryptic enzyme is transported to the stratum with AD in a cohort of 99 patients and 189 controls. corneum extracellular space via lamellar bodies. J Invest Neverthe-less, patients with the AACCAACC allele Dermatol. 1995 May;104(5):819-23 have increased KLK7 protease activity resulting in Franzke CW, Baici A, Bartels J, Christophers E, Wiedow O. premature breakdown of corneodesmosomes, and Antileukoprotease inhibits stratum corneum chymotryptic leading to impairment of the epidermal barrier. enzyme. Evidence for a regulative function in desquamation. J Furthermore, acute eczematous lesions and clini-cally Biol Chem. 1996 Sep 6;271(36):21886-90 unaffected skin can further increase produc-tion of Sondell B, Dyberg P, Anneroth GK, Ostman PO, Egelrud T. KLK7 and epidermal barrier functions are damaged Association between expression of stratum corneum through environmental interactions, such as washing chymotryptic enzyme and pathological keratinization in human with soap and detergents, or long-term application of oral mucosa. Acta Derm Venereol. 1996 May;76(3):177-81 corticosteroids. A combination of the above factors Nylander-Lundqvist E, Egelrud T. Formation of active IL-1 beta leads to a defective skin barrier and increases the risk from pro-IL-1 beta catalyzed by stratum corneum chymotryptic of allergen penetration and succeeding inflammatory enzyme in vitro. Acta Derm Venereol. 1997 May;77(3):203-6 reaction. By ELISA, KLK7 levels were found to be Sondell B, Jonsson M, Dyberg P, Egelrud T. In situ evidence elevated in the stratum corneum of AD patients, and that the population of Langerhans cells in normal human epidermis may be heterogeneous. Br J Dermatol. 1997 KLK7 in the serum significantly correlated with May;136(5):687-93 eosinophil counts in the blood of AD patients, Egelrud T.. Stratum Corneum chymotryptic enzyme. In indicative of the body under an allergic condition. Handbook of proteolytic enzymes. Barrett AJ, Rawlings NP Psoriasis and Woessner JF, Editors., Academic, London. 1998; 87-89. Disease Ekholm E, Egelrud T. The expression of stratum corneum chymotryptic enzyme in human anagen hair follicles: further A number of early studies reported that the evidence for its involvement in desquamation-like processes. chymotrypsin-like activity in stratum corneum was Br J Dermatol. 1998 Oct;139(4):585-90 slightly elevated in psoriasis, but KLK7 serum levels Ekholm E, Sondell B, Strandén P, Brattsand M, Egelrud T. did not differ between normal volunteers and patients Expression of stratum corneum chymotryptic enzyme in human with psoriasis. It has been confirmed that KLK7 protein sebaceous follicles. Acta Derm Venereol. 1998 Sep;78(5):343- levels were similar between non-lesional and lesional 7 skin extracts, but increased amounts of desmoglein 1, Ekholm E, Egelrud T. Stratum corneum chymotryptic enzyme plakoglobin and high molecular weight fragments of in psoriasis. Arch Dermatol Res. 1999 Apr;291(4):195-200 desmocollin 1 were detected in the lesional skin, Tanimoto H, Underwood LJ, Shigemasa K, Yan Yan MS, suggesting an involve-ment of other proteases. Clarke J, Parmley TH, O'Brien TJ. The stratum corneum chymotryptic enzyme that mediates shedding and desquamation of skin cells is highly overexpressed in ovarian References tumor cells. Cancer. 1999 Nov 15;86(10):2074-82 Egelrud T. Purification and preliminary characterization of Egelrud T. Desquamation in the stratum corneum. Acta Derm stratum corneum chymotryptic enzyme: a proteinase that may Venereol Suppl (Stockh). 2000;208:44-5 be involved in desquamation. J Invest Dermatol. 1993 Aug;101(2):200-4 Ekholm E, Egelrud T. Expression of stratum corneum chymotryptic enzyme in relation to other markers of epidermal Egelrud T, Régnier M, Sondell B, Shroot B, Schmidt R. differentiation in a skin explant model. Exp Dermatol. 2000 Expression of stratum corneum chymotryptic enzyme in Feb;9(1):65-70 reconstructed human epidermis and its suppression by retinoic acid. Acta Derm Venereol. 1993 Jun;73(3):181-4 Ekholm IE, Brattsand M, Egelrud T. Stratum corneum tryptic enzyme in normal epidermis: a missing link in the Hansson L, Strömqvist M, Bäckman A, Wallbrandt P, Carlstein desquamation process? J Invest Dermatol. 2000 A, Egelrud T. Cloning, expression, and characterization of Jan;114(1):56-63 stratum corneum chymotryptic enzyme. A skin-specific human serine proteinase. J Biol Chem. 1994 Jul 29;269(30):19420-6 Yousef GM, Scorilas A, Magklara A, Soosaipillai A, Diamandis EP. The KLK7 (PRSS6) gene, encoding for the stratum Sondell B, Thornell LE, Stigbrand T, Egelrud T. corneum chymotryptic enzyme is a new member of the human Immunolocalization of stratum corneum chymotryptic enzyme kallikrein gene family - genomic characterization, mapping, in human skin and oral epithelium with monoclonal antibodies: tissue expression and hormonal regulation. Gene. 2000 Aug evidence of a proteinase specifically expressed in keratinizing 22;254(1-2):119-28 squamous epithelia. J Histochem Cytochem. 1994 Apr;42(4):459-65 Shigemasa K, Tanimoto H, Underwood LJ, Parmley TH, Arihiro K, Ohama K, O'Brien TJ. Expression of the protease inhibitor Egelrud T. [New knowledge of the skin's horny layer may antileukoprotease and the serine protease stratum corneum improve understanding of skin diseases]. Nord Med. chymotryptic enzyme (SCCE) is coordinated in ovarian tumors. 1995;110(3):76-8, 87 Int J Gynecol Cancer. 2001 Nov-Dec;11(6):454-61

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Watkinson A, Harding C, Moore A, Coan P. Water modulation Talieri M, Diamandis EP, Gourgiotis D, Mathioudaki K, Scorilas of stratum corneum chymotryptic enzyme activity and A. Expression analysis of the human kallikrein 7 (KLK7) in desquamation. Arch Dermatol Res. 2001 Sep;293(9):470-6 breast tumors: a new potential biomarker for prognosis of breast carcinoma. Thromb Haemost. 2004 Jan;91(1):180-6 Hansson L, Bäckman A, Ny A, Edlund M, Ekholm E, Ekstrand Hammarström B, Törnell J, Wallbrandt P, Wennbo H, Egelrud Tian X, Shigemasa K, Hirata E, Gu L, Uebaba Y, Nagai N, T. Epidermal overexpression of stratum corneum chymotryptic O'Brien TJ, Ohama K. Expression of human kallikrein 7 enzyme in mice: a model for chronic itchy dermatitis. J Invest (hK7/SCCE) and its inhibitor antileukoprotease (ALP/SLPI) in Dermatol. 2002 Mar;118(3):444-9 uterine endocervical glands and in cervical adenocarcinomas. Oncol Rep. 2004 Nov;12(5):1001-6 Dong Y, Kaushal A, Brattsand M, Nicklin J, Clements JA. Differential splicing of KLK5 and KLK7 in epithelial ovarian Vasilopoulos Y, Cork MJ, Murphy R, Williams HC, Robinson cancer produces novel variants with potential as cancer DA, Duff GW, Ward SJ, Tazi-Ahnini R. Genetic association biomarkers. Clin Cancer Res. 2003 May;9(5):1710-20 between an AACC insertion in the 3'UTR of the stratum corneum chymotryptic enzyme gene and atopic dermatitis. J Johnson B, Horn T, Sander C, Kohler S, R Smoller B. Invest Dermatol. 2004 Jul;123(1):62-6 Expression of stratum corneum chymotryptic enzyme in ichthyoses and squamoproliferative processes. J Cutan Pathol. Bondurant KL, Crew MD, Santin AD, O'Brien TJ, Cannon MJ. 2003 Jul;30(6):358-62 Definition of an immunogenic region within the ovarian tumor antigen stratum corneum chymotryptic enzyme. Clin Cancer Komatsu N, Takata M, Otsuki N, Toyama T, Ohka R, Takehara Res. 2005 May 1;11(9):3446-54 K, Saijoh K. Expression and localization of tissue kallikrein mRNAs in human epidermis and appendages. J Invest Brattsand M, Stefansson K, Lundh C, Haasum Y, Egelrud T. A Dermatol. 2003 Sep;121(3):542-9 proteolytic cascade of kallikreins in the stratum corneum. J Invest Dermatol. 2005 Jan;124(1):198-203 Kyriakopoulou LG, Yousef GM, Scorilas A, Katsaros D, Massobrio M, Fracchioli S, Diamandis EP. Prognostic value of Descargues P, Deraison C, Bonnart C, Kreft M, Kishibe M, quantitatively assessed KLK7 expression in ovarian cancer. Ishida-Yamamoto A, Elias P, Barrandon Y, Zambruno G, Clin Biochem. 2003 Mar;36(2):135-43 Sonnenberg A, Hovnanian A. Spink5-deficient mice mimic Netherton syndrome through degradation of desmoglein 1 by Ny A, Egelrud T. Transgenic mice over-expressing a serine epidermal protease hyperactivity. Nat Genet. 2005 protease in the skin: evidence of interferon gamma- Jan;37(1):56-65 independent MHC II expression by epidermal keratinocytes. 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perspectives on epidermal barrier dysfunction in atopic KLK 7 and FLG genotypes in a French atopic dermatitis cohort. dermatitis: gene-environment interactions. J Allergy Clin Acta Derm Venereol. 2007;87(6):499-505 Immunol. 2006 Jul;118(1):3-21; quiz 22-3 Johnson SK, Ramani VC, Hennings L, Haun RS. Kallikrein 7 Descargues P, Deraison C, Prost C, Fraitag S, Mazereeuw- enhances pancreatic cancer cell invasion by shedding E- Hautier J, D'Alessio M, Ishida-Yamamoto A, Bodemer C, cadherin. Cancer. 2007 May 1;109(9):1811-20 Zambruno G, Hovnanian A. Corneodesmosomal cadherins are preferential targets of stratum corneum trypsin- and Komatsu N, Saijoh K, Kuk C, Liu AC, Khan S, Shirasaki F, chymotrypsin-like hyperactivity in Netherton syndrome. J Invest Takehara K, Diamandis EP. Human tissue kallikrein Dermatol. 2006 Jul;126(7):1622-32 expression in the stratum corneum and serum of atopic dermatitis patients. Exp Dermatol. 2007 Jun;16(6):513-9 Galliano MF, Toulza E, Gallinaro H, Jonca N, Ishida- Yamamoto A, Serre G, Guerrin M. A novel protease inhibitor of Komatsu N, Saijoh K, Kuk C, Shirasaki F, Takehara K, the alpha2-macroglobulin family expressed in the human Diamandis EP. Aberrant human tissue kallikrein levels in the epidermis. J Biol Chem. 2006 Mar 3;281(9):5780-9 stratum corneum and serum of patients with psoriasis: dependence on phenotype, severity and therapy. Br J He QC, Tavakkol A, Wietecha K, Begum-Gafur R, Ansari SA, Dermatol. 2007 May;156(5):875-83 Polefka T. Effects of environmentally realistic levels of ozone on stratum corneum function. Int J Cosmet Sci. 2006 Pampalakis G, Sotiropoulou G. Tissue kallikrein proteolytic Oct;28(5):349-57 cascade pathways in normal physiology and cancer. Biochim Biophys Acta. 2007 Sep;1776(1):22-31 Holzscheiter L, Biermann JC, Kotzsch M, Prezas P, Farthmann J, Baretton G, Luther T, Tjan-Heijnen VC, Talieri M, Schmitt M, Shaw JL, Diamandis EP. Distribution of 15 human kallikreins in Sweep FC, Span PN, Magdolen V. Quantitative reverse tissues and biological fluids. Clin Chem. 2007 Aug;53(8):1423- transcription-PCR assay for detection of mRNA encoding full- 32 length human tissue kallikrein 7: prognostic relevance of KLK7 Yoon H, Laxmikanthan G, Lee J, Blaber SI, Rodriguez A, mRNA expression in breast cancer. Clin Chem. 2006 Kogot JM, Scarisbrick IA, Blaber M. Activation profiles and Jun;52(6):1070-9 regulatory cascades of the human kallikrein-related Prezas P, Arlt MJ, Viktorov P, Soosaipillai A, Holzscheiter L, peptidases. J Biol Chem. 2007 Nov 2;282(44):31852-64 Schmitt M, Talieri M, Diamandis EP, Krüger A, Magdolen V. Zakabunin AI, Mishukova OV, Khrapov EA, Sergeichev DS, Overexpression of the human tissue kallikrein genes KLK4, 5, Boiarskikh UA, Sverdlov ED, Filipenko ML. [Cloning and 6, and 7 increases the malignant phenotype of ovarian cancer expression of the gene of chymotrypsin-like protease of human cells. Biol Chem. 2006 Jun;387(6):807-11 kallikrein-7 in Escherichia coli and isolation of recombinant Prezas P, Scorilas A, Yfanti C, Viktorov P, Agnanti N, protein]. Mol Gen Mikrobiol Virusol. 2007;(2):21-5 Diamandis E, Talieri M. The role of human tissue kallikreins 7 Debela M, Beaufort N, Magdolen V, Schechter NM, Craik CS, and 8 in intracranial malignancies. Biol Chem. 2006 Schmitt M, Bode W, Goettig P. Structures and specificity of the Dec;387(12):1607-12 human kallikrein-related peptidases KLK 4, 5, 6, and 7. Biol Shan SJ, Scorilas A, Katsaros D, Rigault de la Longrais I, Chem. 2008 Jun;389(6):623-32 Massobrio M, Diamandis EP. Unfavorable prognostic value of Dong Y, Matigian N, Harvey TJ, Samaratunga H, Hooper JD, human kallikrein 7 quantified by ELISA in ovarian cancer Clements JA. Tissue-specific promoter utilisation of the cytosols. Clin Chem. 2006 Oct;52(10):1879-86 kallikrein-related peptidase genes, KLK5 and KLK7, and Tan OL, Whitbread AK, Clements JA, Dong Y. Kallikrein- cellular localisation of the encoded proteins suggest roles in related peptidase (KLK) family mRNA variants and protein exocrine pancreatic function. Biol Chem. 2008 Feb;389(2):99- isoforms in hormone-related cancers: do they have a function? 109 Biol Chem. 2006 Jun;387(6):697-705 Eissa A, Diamandis EP. Human tissue kallikreins as Yamasaki K, Schauber J, Coda A, Lin H, Dorschner RA, promiscuous modulators of homeostatic skin barrier functions. Schechter NM, Bonnart C, Descargues P, Hovnanian A, Gallo Biol Chem. 2008 Jun;389(6):669-80 RL. Kallikrein-mediated proteolysis regulates the antimicrobial Fernández IS, Ständker L, Mägert HJ, Forssmann WG, effects of cathelicidins in skin. FASEB J. 2006 Giménez-Gallego G, Romero A. Crystal structure of human Oct;20(12):2068-80 epidermal kallikrein 7 (hK7) synthesized directly in its native Debela M, Hess P, Magdolen V, Schechter NM, Steiner T, state in E. coli: insights into the atomic basis of its inhibition by Huber R, Bode W, Goettig P. Chymotryptic specificity LEKTI domain 6 (LD6). J Mol Biol. 2008 Apr 11;377(5):1488-97 determinants in the 1.0 A structure of the zinc-inhibited human Kiyohara C, Tanaka K, Miyake Y. Genetic susceptibility to tissue kallikrein 7. Proc Natl Acad Sci U S A. 2007 Oct atopic dermatitis. Allergol Int. 2008 Mar;57(1):39-56 9;104(41):16086-91 Oikonomopoulou K, Li L, Zheng Y, Simon I, Wolfert RL, Valik Deraison C, Bonnart C, Lopez F, Besson C, Robinson R, D, Nekulova M, Simickova M, Frgala T, Diamandis EP. Jayakumar A, Wagberg F, Brattsand M, Hachem JP, Prediction of ovarian cancer prognosis and response to Leonardsson G, Hovnanian A. LEKTI fragments specifically chemotherapy by a serum-based multiparametric biomarker inhibit KLK5, KLK7, and KLK14 and control desquamation panel. Br J Cancer. 2008 Oct 7;99(7):1103-13 through a pH-dependent interaction. Mol Biol Cell. 2007 Sep;18(9):3607-19 Planque C, Li L, Zheng Y, Soosaipillai A, Reckamp K, Chia D, Diamandis EP, Goodglick L. A multiparametric serum kallikrein Fernández IS, Ständker L, Forssmann WG, Giménez-Gallego panel for diagnosis of non-small cell lung carcinoma. Clin G, Romero A. Crystallization and preliminary crystallographic Cancer Res. 2008 Mar 1;14(5):1355-62 studies of human kallikrein 7, a serine protease of the multigene kallikrein family. Acta Crystallogr Sect F Struct Biol Psyrri A, Kountourakis P, Scorilas A, Markakis S, Camp R, Cryst Commun. 2007 Aug 1;63(Pt 8):669-72 Kowalski D, Diamandis EP, Dimopoulos MA. Human tissue kallikrein 7, a novel biomarker for advanced ovarian carcinoma Hubiche T, Ged C, Benard A, Léauté-Labrèze C, McElreavey using a novel in situ quantitative method of protein expression. K, de Verneuil H, Taïeb A, Boralevi F. Analysis of SPINK 5, Ann Oncol. 2008 Jul;19(7):1271-7

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Ramani VC, Haun RS. Expression of kallikrein 7 diminishes previously identified serine peptidase inhibitor Kazal type 5 pancreatic cancer cell adhesion to vitronectin and enhances (SPINK5), kallikrein-related peptidase 7 (KLK7), and filaggrin urokinase-type plasminogen activator receptor shedding. (FLG) polymorphisms to eczema risk. J Allergy Clin Immunol. Pancreas. 2008 Nov;37(4):399-404 2008 Sep;122(3):560-8.e4 Ramani VC, Haun RS. The extracellular matrix protein Xuan Q, Yang X, Mo L, Huang F, Pang Y, Qin M, Chen Z, He fibronectin is a substrate for kallikrein 7. Biochem Biophys Res M, Wang Q, Mo ZN. Expression of the serine protease Commun. 2008 May 16;369(4):1169-73 kallikrein 7 and its inhibitor antileukoprotease is decreased in prostate cancer. Arch Pathol Lab Med. 2008 Ramani VC, Hennings L, Haun RS. Desmoglein 2 is a Nov;132(11):1796-801 substrate of kallikrein 7 in pancreatic cancer. BMC Cancer. 2008 Dec 17;8:373 Nin M, Katoh N, Kokura S, Handa O, Yoshikawa T, Kishimoto S. Dichotomous effect of ultraviolet B on the expression of Simon M, Tazi-Ahnini R, Jonca N, Caubet C, Cork MJ, Serre corneodesmosomal enzymes in human epidermal G. Alterations in the desquamation-related proteolytic cleavage keratinocytes. J Dermatol Sci. 2009 Apr;54(1):17-24 of corneodesmosin and other corneodesmosomal proteins in psoriatic lesional epidermis. Br J Dermatol. 2008 Jul;159(1):77- Pettus JR, Johnson JJ, Shi Z, Davis JW, Koblinski J, Ghosh S, 85 Liu Y, Ravosa MJ, Frazier S, Stack MS. Multiple kallikrein (KLK 5, 7, 8, and 10) expression in squamous cell carcinoma of the Singh J, Naran A, Misso NL, Rigby PJ, Thompson PJ, Bhoola oral cavity. Histol Histopathol. 2009 Feb;24(2):197-207 KD. Expression of kallikrein-related peptidases (KRP/hK5, 7, 6, 8) in subtypes of human lung carcinoma. Int Talieri M, Mathioudaki K, Prezas P, Alexopoulou DK, Immunopharmacol. 2008 Feb;8(2):300-6 Diamandis EP, Xynopoulos D, Ardavanis A, Arnogiannaki N, Scorilas A. Clinical significance of kallikrein-related peptidase 7 Weidinger S, Baurecht H, Wagenpfeil S, Henderson J, Novak (KLK7) in colorectal cancer. Thromb Haemost. 2009 N, Sandilands A, Chen H, Rodriguez E, O'Regan GM, Watson Apr;101(4):741-7 R, Liao H, Zhao Y, Barker JN, Allen M, Reynolds N, Meggitt S, Northstone K, Smith GD, Strobl C, This article should be referenced as such: Stahl C, Kneib T, Klopp N, Bieber T, Behrendt H, Palmer CN, Dong Y, Lai J, Clements JA. KLK7 (kallikrein-related peptidase Wichmann HE, Ring J, Illig T, McLean WH, Irvine AD. Analysis 7). Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4):389- of the individual and aggregate genetic contributions of 396.

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Gene Section Mini Review

LATS1 (LATS, large tumor suppressor, homolog 1 (Drosophila)) Stacy Visser, Xiaolong Yang Department of Pathology and Molecular Medicine, Queen's University Kingston, ON, Canada (SV, XY)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/LATS1ID41127ch6q25.html DOI: 10.4267/2042/44735 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

domain lie two conserved residues, Ser909 and Identity Thr1079, which are phosphorylated upon activa-tion. Other names: EC 2.7.11.1; WARTS; h-warts; wts The N-terminus of LATS1 contains two PPxY motifs, HGNC (Hugo): LATS1 which bind to WW-domain transcriptional co- activators TAZ and YAP, as well as a protein binding Location: 6q25.1 domain (PBD) that has been shown to bind to MOB1 Local order: KATNA1 - LATS1 - LOC645967 - and cytoskeletal proteins LIMK1 and Zyxin. In NUP43 addition, an ubiquitin binding domain (UBA) and a proline-rich region (P-stretch) have been identified in DNA/RNA the N-terminal region of LATS1, although the functional significance of these domains has not yet Description been determined. The LATS1 gene contains 8 exons, ranging in size from 106 bp to 1513 bp. The mRNA transcript spans Expression 4756 bp. LATS1 is ubiquitously expressed. Transcription Localisation An alternatively spliced variant was identified in LATS1 is primarily localized within the cytoplasm, vertebrate retina and testis. This smaller variant has however, LATS1 possesses functions that require its deletions of exon 6, 7, and 8. The functional translocation to the nucleus. The mechanism of significance of this splice variant is not known. translocation is not yet understood. Protein Function As a tumor suppressor, LATS1 functions as a key Description regulator of cell cycle progression, apoptosis, and cell LATS1 is a highly conserved Ser/Thr kinase that migration. As cell cycle regulator, LATS1 is able to belongs to the AGC (Protein kinase A/G/C) family of modulate multiple aspects of the cell cycle, including protein kinases. Within the C-terminal kinase G2/M arrest, mitotic exit, activation of

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 397 LATS1 (LATS, large tumor suppressor, homolog 1 (Drosophila)) Visser S, Yang X

the G1 tetraploidy checkpoint, and regulation of cytokinesis. LATS1 also promotes apoptosis by Implicated in inducing expression of pro-apoptotic proteins BAX, Cancers caspase 3 and tumor suppressor p53. Finally, loss of LATS1 has been shown to enhance the rate of cell Disease migration. There is increasing evidence that LATS1 is downregulated in a variety of human tumor types. To HIPPO-LATS Signaling Pathway: LATS1 is a key date, loss of LATS1 has been found in soft tissue player in the conserved Hippo-LATS signaling sarcomas, particularly myxoid liposarcoma, pathway. Components of this pathway include the leiomyosarcoma, and malignant fibrous histocyto-mas, adaptor proteins WW45 and MOB, which aid in as well as ovarian cancer, breast cancer, acute bringing LATS in contact with the kinase MST1/ lymphoblastic leukemia, and astrocytoma. Prima-rily MST2. MST1/2 can then phosphorylate and activate downregulation of LATS1 expression is due to LATS1. Upstream of MST1/2 lay FERM domain hypermethylation of the promoter region. proteins Willin/FRMD6 and Merlin/NF2, as well as a protocadherin FAT4. The molecular mechanisms Prognosis regulating this pathway have not yet been delineated in Decreased LATS1 expression in breast cancer is a mammalian system. Down-stream of LATS1 lay two associated with large tumor size, high lymph node transcriptional co-activators, YAP and TAZ, which are metastasis, and estrogen and progesterone nega-tivity. phosphoryla-ted and inhibited by LATS1, thereby Thus, loss of LATS1 is associated with poor prognosis sequestering them in the cytoplasm. Finally, YAP and in breast cancer. Furthermore, loss of LATS1 TAZ have been shown to act through the TEAD/TEF expression, in addition to other genes, can be used as a family of transcription factors to modulate the prognostic factor in predicting disease-free survival for expression of a variety of genes (See figure below). acute lymphoblastic leukemia. Function-ally, the Hippo-LATS pathway has been implicated in cell proliferation, apoptosis, the epithelial References mesen-chymal transition, and cell migration. Tao W, Zhang S, Turenchalk GS, Stewart RA, St John MA, Chen W, Xu T. Human homologue of the Drosophila Homology melanogaster lats tumour suppressor modulates CDC2 The LATS1 kinase domain is conserved across species. activity. Nat Genet. 1999 Feb;21(2):177-81 Human homologs include LATS2 and the nuclear Turenchalk GS, St John MA, Tao W, Xu T. The role of lats in Dbf2-related kinases, NDR1 and NDR2. cell cycle regulation and tumorigenesis. Biochim Biophys Acta. 1999 Oct 29;1424(2-3):M9-M16 Mutations Hirota T, Morisaki T, Nishiyama Y, Marumoto T, Tada K, Hara T, Masuko N, Inagaki M, Hatakeyama K, Saya H. Zyxin, a Note regulator of actin filament assembly, targets the mitotic There have been no reports of mutations in LATS1. apparatus by interacting with h-warts/LATS1 tumor suppressor. J Cell Biol. 2000 May 29;149(5):1073-86

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 398 LATS1 (LATS, large tumor suppressor, homolog 1 (Drosophila)) Visser S, Yang X

Yang X, Li DM, Chen W, Xu T. Human homologue of Hergovich A, Schmitz D, Hemmings BA. The human tumour Drosophila lats, LATS1, negatively regulate growth by inducing suppressor LATS1 is activated by human MOB1 at the G(2)/M arrest or apoptosis. Oncogene. 2001 Oct membrane. Biochem Biophys Res Commun. 2006 Jun 4;20(45):6516-23 23;345(1):50-8 Hisaoka M, Tanaka A, Hashimoto H. Molecular alterations of h- Jiang Z, Li X, Hu J, Zhou W, Jiang Y, Li G, Lu D. Promoter warts/LATS1 tumor suppressor in human soft tissue sarcoma. hypermethylation-mediated down-regulation of LATS1 and Lab Invest. 2002 Oct;82(10):1427-35 LATS2 in human astrocytoma. Neurosci Res. 2006 Dec;56(4):450-8 Iida S, Hirota T, Morisaki T, Marumoto T, Hara T, Kuninaka S, Honda S, Kosai K, Kawasuji M, Pallas DC, Saya H. Tumor Hao Y, Chun A, Cheung K, Rashidi B, Yang X. Tumor suppressor WARTS ensures genomic integrity by regulating suppressor LATS1 is a negative regulator of oncogene YAP. J both mitotic progression and G1 tetraploidy checkpoint Biol Chem. 2008 Feb 29;283(9):5496-509 function. Oncogene. 2004 Jul 8;23(31):5266-74 Lei QY, Zhang H, Zhao B, Zha ZY, Bai F, Pei XH, Zhao S, Roman-Gomez J, Jimenez-Velasco A, Castillejo JA, Agirre X, Xiong Y, Guan KL. TAZ promotes cell proliferation and Barrios M, Navarro G, Molina FJ, Calasanz MJ, Prosper F, epithelial-mesenchymal transition and is inhibited by the hippo Heiniger A, Torres A. Promoter hypermethylation of cancer- pathway. Mol Cell Biol. 2008 Apr;28(7):2426-36 related genes: a strong independent prognostic factor in acute lymphoblastic leukemia. Blood. 2004 Oct 15;104(8):2492-8 Zeng Q, Hong W. The emerging role of the hippo pathway in cell contact inhibition, organ size control, and cancer Yang X, Yu K, Hao Y, Li DM, Stewart R, Insogna KL, Xu T. development in mammals. Cancer Cell. 2008 Mar;13(3):188-92 LATS1 tumour suppressor affects cytokinesis by inhibiting LIMK1. Nat Cell Biol. 2004 Jul;6(7):609-17 Zhang J, Smolen GA, Haber DA. Negative regulation of YAP by LATS1 underscores evolutionary conservation of the Chan EH, Nousiainen M, Chalamalasetty RB, Schäfer A, Nigg Drosophila Hippo pathway. Cancer Res. 2008 Apr EA, Silljé HH. The Ste20-like kinase Mst2 activates the human 15;68(8):2789-94 large tumor suppressor kinase Lats1. Oncogene. 2005 Mar 17;24(12):2076-86 This article should be referenced as such: Takahashi Y, Miyoshi Y, Takahata C, Irahara N, Taguchi T, Visser S, Yang X. LATS1 (LATS, large tumor suppressor, Tamaki Y, Noguchi S. Down-regulation of LATS1 and LATS2 homolog 1 (Drosophila)). Atlas Genet Cytogenet Oncol mRNA expression by promoter hypermethylation and its Haematol. 2010; 14(4):397-399. association with biologically aggressive phenotype in human breast cancers. Clin Cancer Res. 2005 Feb 15;11(4):1380-5

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 399 Atlas of Genetics and Cytogenetics

in Oncology and Haematology OPEN ACCESS JOURNAL AT INIST-CNRS

Gene Section Review

NOL3 (nucleolar protein 3 (apoptosis repressor with CARD domain)) Gloria Kung, Wendy McKimpson, Richard N Kitsis Department of Medicine, Department of Cell Biology, Montefiore-Einstein Center for Cardiovascular Research, Albert Einstein Cancer Center, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461 USA (GK, WMK, RNK)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/NOL3ID41552ch16q22.html DOI: 10.4267/2042/44736 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity names of the putative encoded proteins: ARC (Apoptosis Repressor with CARD) CARD Other names: ARC (Apoptosis Repressor with denotes a Caspase Recruitment Domain. The ARC CARD); CARD2; MYP; NOP; NOP30 protein resides in the cytoplasm and nucleoplasm, not HGNC (Hugo): NOL3 nucleolus. Location: 16q22.1 MYP is an older name for ARC that is currently not Note used. References to this locus as MYC are incorrect and probably represent typographical errors of MYP. The correct name for the locus is NOL3. Sometimes, NOL3 is distinct from any of the myc loci. however, the gene is referred to by

The NOL3 gene is located on the long arm of human chromosome 16. The gene consists of 4 small exons (exons denoted above as thick boxes) and 3 small introns. The translational start site is in exon 2. Alternative splicing occurs between exons 2 and 3. This involves two splice donors separated by 10 nucleotides in exon 2 connecting to a single splice acceptor in exon 3 (Stoss et al., 1999). Because the separation between the splice donors, 10 nucleotides, is not an exact multiple of 3, alternative splicing results in two open reading frames distal to the splice acceptor. Because of this frame shift, the C-terminus of the two encoded proteins differ as do their stop codons, each of which is in exon 4. One transcript is translated into ARC (Apoptosis Repressor with CARD (Caspase Recruitment Domain)) (Koseki et al., 1998). MYP is an earlier name for ARC that is no longer in use (Geertman et al., 1996). The other transcript encodes a putative protein called NOP30 (Nucleolar Protein of 30 kD). ARC and putative NOP30 proteins share a common N-terminus containing the CARD. Their C-termini differ, however, with ARC containing multiple P/E repeats (acidic) and putative NOP30 containing R/S repeats (basic). While ARC transcripts are present in a variety of human and mouse cell types, NOP30 transcripts are present in human, but not mouse (L. Wu and R. Kitsis, unpublished). Endogenous ARC protein resides in the cytoplasm and nucleoplasm of certain human and mouse cell types (discussed below). In contrast, the existence of endogenous NOP30 protein has not been demonstrated in any cell type of any species. When the cDNA encoding NOP30 is exogenously expressed, the encoded protein is in the nucleolus and nucleoplasm (Stoss et al., 1999).

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 400 NOL3 (nucleolar protein 3 (apoptosis repressor with CARD domain)) Kung G, et al.

NOP30. In some species, alternative splicing gives rise 2001). ARC protein is also markedly increased in to a transcript encoding a putative protein NOP30, primary human epithelial cancers of the breast, colon, rather than ARC. When the cDNA for NOP30 is ovary, and cervix (Mercier et al., 2005; Mercier et al., expressed exogenously, the resulting protein is 2008). NOP30 transcripts are present in some human predominantly nucleolar - hence the origin of the gene cell types but have not been detected in mouse cells. name: nucleolar protein 3. Impo-rtantly, however, Endogenous NOP30 protein has not been demonstrated endogenous NOP30 protein has not been demonstrated in cells of any species. in any cells of any species. Localisation DNA/RNA Endogenous ARC protein is present in the cytoplasm and nucleoplasm (Mercier et al., 2005). As above, the Description localization of endogenous NOP30 protein has not been The NOL3 gene is located on human chromosome investigated. Exogenously expressed NOP30 protein 16q21-23. The gene contains 4 exons and 3 introns localizes in the nucleolus and nucleoplasm. spanning 1757 bp. Function Transcription The function of endogenous NOP30 is not known. The coordinate of the first nucleotide of exon 1 is Exogenous NOP30 interacts with SFRS9/SRp30C and 65,765,371 bp from pter, and that of the last nucleotide NPM1 and may influence splicing (Stoss et al., 1999). of exon 4 is 65,767,127 bp. Alternative splicing takes ARC is an endogenous inhibitor of apoptosis that is place between exons 2 and 3. In exon 2, the splice unique in its ability to antagonize both the extrinsic donor of the NOP30 transcript is 10 bp upstream of the (death receptor) and the intrinsic (mitochondria/ER) splice donor of the ARC transcript. Both transcripts use death pathways (Nam et al., 2004; Gustafsson et al., a common splice acceptor in exon 3. 2004; Koseki et al., 1998). ARC inhibits the extrinsic pathway by interfering with DISC (Death Inducing Protein Signaling Complex) formation. This is accomplished by the direct interaction of the ARC CARD with the Note death domains (DD) of Fas and FADD, and with the The start of translation is in exon 2 (prior to the death effector domain (DED) of procaspase-8. These alternative splice donors). Alternative splicing causes a death-fold interactions are novel in that they are frame shift resulting in transcripts encoding proteins heterotypic in contrast to the usual homotypic death- with different C-termini and separate stop codons in fold interactions. ARC inhibits the intrinsic pathway exon 4. The stop codon for ARC is 43 bp upstream of through at least two mechanisms. First, the direct that of NOP30. interaction between the ARC CARD and the C- terminus of Bax inhibits death stimulus-induced Bax conformational activa-tion and translocation to the mitochondria. Second, direct interaction between the ARC C-terminal domain with the p53 tetramerization domain inhibits p53 tetramerization (Foo et al., PNAS, 2007). This, in turn, disables p53 transcriptional Alternatively spliced transcripts of NOL3 lead to two different function and exposes a p53 nuclear export signal that proteins, ARC (blue) and NOP30 (red). These proteins each relocates p53 to the cytoplasm. contain an N-terminal CARD (first 95 amino acids identical), but have different C-termini. The C-terminus of ARC is rich in Nothing is known about the regulation of NOP30. prolines and glutamic acids, whereas the C-terminus of NOP30 The regulation of ARC is complex. ARC protein is rich in serines and arginines. abundance decreases rapidly and dramatically in Description response to hypoxia and oxidative stress (e.g. ischemia-

Human ARC protein contains 208 amino acids with M r reperfusion) (Ekhterae et al., 1999; Neuss et al., 2001; 22,629 Da. The protein usually runs at a slower Nam et al., 2007). These decreases result from mobility on SDS-PAGE most likely due to the increased degradation of ARC protein via the ubiquitin- enrichment of proline residues in the C-terminal proteasomal pathway (Nam et al., 2007). The E3 ligase MDM2 may play a role in ARC degradation in this domain. NOP30 contains 219 amino acids with M r 24,327 Da. scenario (Foo et al., JBC, 2007), but this role is probably indirect (L. Wu and R. Kitsis, unpublished Expression data). Decreases in ARC protein abundance in response Under normal conditions, ARC mRNA and protein is to hypoxia appear to be regulated by p53 repression of present predominantly in cardiac myocytes, skeletal nol3 transcription (Li et al., 2008). Apart from ARC myocytes, and neurons (Koseki et al., 1998; Abmayr et protein abundance, the activity of ARC is also al., 2004; Geertman et al., 1996; Engidawork et al., regulated

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 401 NOL3 (nucleolar protein 3 (apoptosis repressor with CARD domain)) Kung G, et al.

Regulation of the extrinsic (death receptor) and intrinsic (mitochondria/ER) apoptosis pathways by ARC. Not shown are ARC interactions with and regulation of p53. post-translationally: dephosphorylation of threonine damage during myocardial infarction. Endogenous 149 decreases the anti-apoptotic activity of ARC (Tan ARC protein undergoes rapid protea-somal degradation et al., 2008). during myocardial ischemia-reperfusion (Nam et al., 2007). This decrease in ARC abundance is causally Homology linked with the subsequent cell death (Nam et al., ARC is highly conserved among mammals. There is 2004). Accordingly, transgenic overexpression of ARC approximately 85% identity both at the amino acid and in vivo decreases the size of myocardial infarctions the nucleotide level among human, rat, mouse, dog, and (Gustafsson et al., 2002; Pyo et al., 2008; S. Jha and R. bovine ARC. Interestingly, an ARC homolog has yet to Kitsis, unpublished data). As would be predicted, be identified in Danio rerio, Drosophila melanogaster, knockout of ARC has been reported to result in larger or Caenorhabditis elegans. infarcts (Donath et al., 2006). However, the aforementioned knockout studies were performed on Implicated in only small numbers of mice on a mixed genetic background. Subsequent knockout studies involving Epithelial cancers large numbers of mice on several pure genetic Disease backgrounds have not demonstrated larger infarcts in Increased levels of ARC protein have been observed in ARC-/- mice subjected to ische-mia-reperfusion (J. the epithelium of primary human breast, colon, ovarian, Saurabh, S. Y. Ji, and R. Kitsis, unpublished data). This and cervical cancers (Mercier et al., 2005; Mercier et is probably due to the dramatic rapid degradation of al., 2008). Increased levels of both ARC RNA and ARC protein during reperfusion even in wild type mice protein have been observed in renal cell carcinoma (see above). (Heikaus et al., 2008). Heart failure Prognosis Prognosis ARC overexpression in a breast cancer cell line ARC protein levels decrease during heart failure. increases resistance to chemotherapy and radiation Moreover, knockout of ARC exacerbates patholo-gical (Mercier et al., 2005; Wang et al., 2009). In a mela- cardiac remodeling in response to hemody-namic noma cell line, ARC overexpression causes increased overload, a model of heart failure (Donath et al., 2006). resistance to endoplasmic reticulum stress-induced caspase-8 activation (Chen et al., 2008). Neuropathology (several individual Myocardial infarction, myocardial entities) ischemia-reperfusion Prognosis Prognosis The protein level of ARC is increased in the frontal cortex of patients with Alzheimer's disease ARC plays an important role in regulating heart muscle

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 402 NOL3 (nucleolar protein 3 (apoptosis repressor with CARD domain)) Kung G, et al.

(Engidawork et al., 2001). During ischemic injury of nonhomotypic death-fold interactions. Mol Cell. 2004 Sep the brain, there is a decrease in ARC protein in 24;15(6):901-12 hippocampal neurons (Hong et al., 2003). Other studies Mercier I, Vuolo M, Madan R, Xue X, Levalley AJ, Ashton AW, have also shown that caloric restriction increases Jasmin JF, Czaja MT, Lin EY, Armstrong RC, Pollard JW, Kitsis RN. ARC, an apoptosis suppressor limited to terminally expression of ARC in the brain (Shelke et al., 2003). differentiated cells, is induced in human breast cancer and confers chemo- and radiation-resistance. Cell Death Differ. Breakpoints 2005 Jun;12(6):682-6 Note Donath S, Li P, Willenbockel C, Al-Saadi N, Gross V, Willnow T, Bader M, Martin U, Bauersachs J, Wollert KC, Dietz R, von Not known. Harsdorf R. Apoptosis repressor with caspase recruitment domain is required for cardioprotection in response to References biomechanical and ischemic stress. Circulation. 2006 Mar 7;113(9):1203-12 Geertman R, McMahon A, Sabban EL. Cloning and Foo RS, Chan LK, Kitsis RN, Bennett MR. Ubiquitination and characterization of cDNAs for novel proteins with glutamic degradation of the anti-apoptotic protein ARC by MDM2. J Biol acid-proline dipeptide tandem repeats. Biochim Biophys Acta. Chem. 2007 Feb 23;282(8):5529-35 1996 May 2;1306(2-3):147-52 Foo RS, Nam YJ, Ostreicher MJ, Metzl MD, Whelan RS, Peng Koseki T, Inohara N, Chen S, Núñez G. ARC, an inhibitor of CF, Ashton AW, Fu W, Mani K, Chin SF, Provenzano E, Ellis I, apoptosis expressed in skeletal muscle and heart that interacts Figg N, Pinder S, Bennett MR, Caldas C, Kitsis RN. Regulation selectively with caspases. Proc Natl Acad Sci U S A. 1998 Apr of p53 tetramerization and nuclear export by ARC. Proc Natl 28;95(9):5156-60 Acad Sci U S A. 2007 Dec 26;104(52):20826-31 Ekhterae D, Lin Z, Lundberg MS, Crow MT, Brosius FC 3rd, Nam YJ, Mani K, Wu L, Peng CF, Calvert JW, Foo RS, Núñez G. ARC inhibits cytochrome c release from Krishnamurthy B, Miao W, Ashton AW, Lefer DJ, Kitsis RN. mitochondria and protects against hypoxia-induced apoptosis The apoptosis inhibitor ARC undergoes ubiquitin-proteasomal- in heart-derived H9c2 cells. Circ Res. 1999 Dec 9;85(12):e70-7 mediated degradation in response to death stimuli: Stoss O, Schwaiger FW, Cooper TA, Stamm S. Alternative identification of a degradation-resistant mutant. J Biol Chem. splicing determines the intracellular localization of the novel 2007 Feb 23;282(8):5522-8 nuclear protein Nop30 and its interaction with the splicing Chen LH, Jiang CC, Watts R, Thorne RF, Kiejda KA, Zhang factor SRp30c. J Biol Chem. 1999 Apr 16;274(16):10951-62 XD, Hersey P. Inhibition of endoplasmic reticulum stress- Engidawork E, Gulesserian T, Yoo BC, Cairns N, Lubec G. induced apoptosis of melanoma cells by the ARC protein. Alteration of caspases and apoptosis-related proteins in brains Cancer Res. 2008 Feb 1;68(3):834-42 of patients with Alzheimer's disease. Biochem Biophys Res Heikaus S, Kempf T, Mahotka C, Gabbert HE, Ramp U. Commun. 2001 Feb 16;281(1):84-93 Caspase-8 and its inhibitors in RCCs in vivo: the prominent Neuss M, Monticone R, Lundberg MS, Chesley AT, Fleck E, role of ARC. Apoptosis. 2008 Jul;13(7):938-49 Crow MT. The apoptotic regulatory protein ARC (apoptosis Li YZ, Lu DY, Tan WQ, Wang JX, Li PF. p53 initiates apoptosis repressor with caspase recruitment domain) prevents oxidant by transcriptionally targeting the antiapoptotic protein ARC. Mol stress-mediated cell death by preserving mitochondrial Cell Biol. 2008 Jan;28(2):564-74 function. J Biol Chem. 2001 Sep 7;276(36):33915-22 Mercier I, Vuolo M, Jasmin JF, Medina CM, Williams M, Gustafsson AB, Sayen MR, Williams SD, Crow MT, Gottlieb Mariadason JM, Qian H, Xue X, Pestell RG, Lisanti MP, Kitsis RA. TAT protein transduction into isolated perfused hearts: RN. ARC (apoptosis repressor with caspase recruitment TAT-apoptosis repressor with caspase recruitment domain is domain) is a novel marker of human colon cancer. Cell Cycle. cardioprotective. Circulation. 2002 Aug 6;106(6):735-9 2008 Jun 1;7(11):1640-7 Hong YM, Jo DG, Lee JY, Chang JW, Nam JH, Noh JY, Koh Pyo JO, Nah J, Kim HJ, Chang JW, Song YW, Yang DK, Jo JY, Jung YK. Down-regulation of ARC contributes to DG, Kim HR, Chae HJ, Chae SW, Hwang SY, Kim SJ, Kim HJ, vulnerability of hippocampal neurons to ischemia/hypoxia. Cho C, Oh CG, Park WJ, Jung YK. Protection of FEBS Lett. 2003 May 22;543(1-3):170-3 cardiomyocytes from ischemic/hypoxic cell death via Drbp1 Shelke RR, Leeuwenburgh C. Lifelong caloric restriction and pMe2GlyDH in cardio-specific ARC transgenic mice. J Biol increases expression of apoptosis repressor with a caspase Chem. 2008 Nov 7;283(45):30707-14 recruitment domain (ARC) in the brain. FASEB J. 2003 Tan WQ, Wang JX, Lin ZQ, Li YR, Lin Y, Li PF. Novel cardiac Mar;17(3):494-6 apoptotic pathway: the dephosphorylation of apoptosis Abmayr S, Crawford RW, Chamberlain JS. Characterization of repressor with caspase recruitment domain by calcineurin. ARC, apoptosis repressor interacting with CARD, in normal Circulation. 2008 Nov 25;118(22):2268-76 and dystrophin-deficient skeletal muscle. Hum Mol Genet. Wang JX, Li Q, Li PF. Apoptosis repressor with caspase 2004 Jan 15;13(2):213-21 recruitment domain contributes to chemotherapy resistance by Gustafsson AB, Tsai JG, Logue SE, Crow MT, Gottlieb RA. abolishing mitochondrial fission mediated by dynamin-related Apoptosis repressor with caspase recruitment domain protects protein-1. Cancer Res. 2009 Jan 15;69(2):492-500 against cell death by interfering with Bax activation. J Biol Chem. 2004 May 14;279(20):21233-8 This article should be referenced as such: Nam YJ, Mani K, Ashton AW, Peng CF, Krishnamurthy B, Kung G, McKimpson W, Kitsis RN. NOL3 (nucleolar protein 3 Hayakawa Y, Lee P, Korsmeyer SJ, Kitsis RN. Inhibition of (apoptosis repressor with CARD domain)). Atlas Genet both the extrinsic and intrinsic death pathways through Cytogenet Oncol Haematol. 2010; 14(4):400-403.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 403 Atlas of Genetics and Cytogenetics

in Oncology and Haematology OPEN ACCESS JOURNAL AT INIST-CNRS

Gene Section Review

NPY (neuropeptide Y) Massimiliano Ruscica, Elena Dozio, Paolo Magni Dipartimento di Endocrinologia, Fisiopatologia e Biologia Applicata, Università degli Studi di Milano, Italy (MR, PM); Dipartimento di Morfologia Umana e Scienze Biomediche "Città Studi", Università degli Studi di Milano, Italy (ED)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/NPYID44438ch7p15.html DOI: 10.4267/2042/44737 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

contains the 5'-UTR, Exon 2 (188 bp) encodes the Identity signal peptide and the main part of the mature NPY Other names: PYY4 sequence, except the last base of arginine codon 35 and HGNC (Hugo): NPY the last tyrosine codon. Exon 3 (82 bp) contains the last base of the arginine codon, the last tyrosine codon from Location: 7p15.1 the mature peptide, the glycine amide donor site, the dibasic cleavage site, and the main portion of C- DNA/RNA terminal flanking peptide of NPY (CPON). Finally, Note Exon 4 (21 + 174 bp) includes the end of CPON and History: NPY, first discovered in 1982 from pig brain, the 3'-UTR. has been characterized as one of the most highly conserved neuroendocrine peptides through-out Protein evolution. Its name derives from the single-letter code Description (Y) for the amino acid (aa) tyrosine, since it contains five tyrosine residues including an amidated C-terminal The coding sequence (291 bp) of NPY gene synthetizes tyrosine residue. Moreover, there is 92% aa sequence the pre-pro NPY (see diagram), a pre-cursor peptide of identity for NPY between the cartilaginous fish, 97 aa residues (size 10.8 kDa), which is cleaved after Torpedo marmorata, and mammals, which are 28-aa, resulting in the pro-NPY, a 69-aa peptide. Pro- separated by an evolutionary distance of more than 400 NPY undergoes clea-vage by the proconverting enzymes (PC)1/3 and/or PC2 at a single dibasic site million years. These evidences indicate that NPY 38 39 presumably has a critical physiological function and (Lys -Arg ), releasing a 30-aa carboxyl terminal that interindi-vidual variation at this locus is likely to peptide, the C-flanking peptide of NPY (CPON), with be minimal. The NPY gene is about 8 kilobases (kb) in so far unknown function and NPY 1-39 , which is further length with four small exons (each, <200 base pairs processed by a carboxypeptidase-like enzyme. The [bp]) separated by three introns of approximately 965, resulting NPY 1-37 is finally amidated at its C-terminal 4300, and 2300 bp. end by the peptidylglycine-amidating monooxygenase (PAM) in a process that removes an additional aa Description (glycine) and produces the mature and biologically NPY encompasses 7,669 bp of DNA on chromosome 7 active peptide NPY 1-36 . Moreover, NPY can be further (7p15.1) between 24290334 and 24298002 from pter. process, by two enzymes, dipeptidyl peptidase IV Transcription (DPPIV) and aminopeptidase P, into NPY 3-36 and NPY 2-36 , respectively. NPY exhibits a three- The NPY RNA transcript contains 4 exons and it is 551 dimensional structure called PP-fold, which is bp, in lenght. In details, Exon 1 (86 bp) characterized by a harpin-like structure and by a type II beta-turn, connecting a polyproline-like type

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 404 NPY (neuropeptide Y) Ruscica M, et al.

A schematic drawing of 97 aa pre-pro NPY which is further process up to the release of the mature and biologically active peptide NPY 1- 36 and the 30-aa CPON peptide.

II helix (residues 14 to 30), thus leading to a close combination of Henle's, intermediate and Meissner's proximity of N- and C-terminal ends of the peptide. plexi) cell bodies are NPY-positive and co-localised with nitric oxide synthase immuno-reactivity. Expression Localisation The 36-aa NPY is one of the most abundant peptides Stored in cytoplasmatic secretory granules within the expressed in numerous brain regions (i.e, cell body and peptidergic nerve terminals. It is secreted hypothalamus, amygdala, hippocampus and cerebral upon specific stimuli. cortex), as well as in the periphery (i.e, liver, heart, spleen, endothelial cells of blood vessels), showing Function both pre- and post-synaptic actions. However, the NPY has been involved in the regulation of a wide adrenal medulla is the primary source of circulating range of physiological central effects like the control of NPY. appetite, body weight homeostasis, the modulation of In details, in the brain, it is synthesized by neurons of reproductive processes, the regula-tion of growth the arcuate (ARC) nucleus and it is active on the hormone secretion. Moreover, it is involved in the paraventricular (PVN), on the dorsomedial (DMN) anxyolytic effect and sedation, in the endogenous nucleus, and on the A1 and C1 areas of the median anticonvulsant activity, in the circadian rhythm preoptic area. In particular, in human, high levels of regulation, in learning and memory, in analgesia and NPY mRNA were found in the dentate gyrus, caudate hyperalgesia. nucleus and putamen. In cerebral cortex the amount of 1) Food Intake NPY-immunoreactivity (NPY-ir) is present at the Being NPY a very potent stimulant of food ingestion, highest concentration in the cingulated and temporal and mainly of carbohydrates, among these NPY- cortices and at the lowest in the occipital lobe. NPY- mediated actions, the major one is its role in the control labeled cells are also found in subcortical white matter, of appetite, body weight homeostasis, and obesity. All whereas this expression is negative in white matter these functions are mediated in humans by different areas away from the cortex. The striatum shows receptor subtypes, so-called Y receptors: Y1-R, Y2-R, heterogeneous levels of NPY mRNA with a higher Y4-R, Y5-R and y6-R (a pseudogene in primates, with expression in the nucleus accumbens and the ventral no shown activity as yet), which all belong to the large region of the caudate. In general, NPY neurons also family of G -protein coupled receptors. coexpress various neurotransmitters. In fact, ARC NPY i sub-population coexpresses the neuropeptide agouti- 2) Cardiovascular function related peptide (AgRP) and the amino acid gamma- NPY has also been described to mediate peripheral aminobutyric acid (GABA), whereas the brain stem effects including the regulation of the cardiova-scular subpopulation coexpresses the adrenergic trans-mitters, system, vasoconstriction, the release of cate- epinephrine and norepinephrine. cholamines by the adrenal medulla and vascular In the periphery, NPY is expressed in the sympathetic motility. About 25% of patients with acute heart failure nervous system, co-stored and co-released with were found to display an increase in circulating NPY norepinephrine, as well as in non-sympathetic neurons levels. NPY has been proposed to be an useful marker in several organs including gastrointestinal tract, for estimating the condition of patients suffering from salivary glands, thyroid gland, pancreas, urogenital cardiac diseases. In fact, plasma NPY system, Schwann cells and heart. NPY is also present immunoreactivity appeared elevated in patients with in cholinergic and non-cholinergic neurons (both acute myocardial ischemia and in those with congestive central and peripheral) with anti-depressant, anti- heart failure. Moreover, NPY appears to enhance convulsant and anti-nociceptive actions. In the diuresis and natriuresis through direct tubular effects mammalian intestine, NPY is present in both myenteric independent of its hemody-namic properties. and submucous neurones that provide an extensive The vasopressive properties of NPY as well as its intrinsic innervation to smooth muscle layers and capacity to potentiate adrenergic responsiveness mucosal targets, respectively. NPY has been detected suggests that this peptide might be beneficial in the in glial cells and in olfactory Schwann cells, thereby management of endotoxic shock. Indeed, in humans potentially affording trophic support. NPY-ir appears experiencing sepsis and septic shock, plasma levels of mainly found in the renal tubules, whereas it seems NPY are significantly increased and NPY-mediated absent in the glomerules. Interestingly, in infant colon, vasoconstriction was shown to be preserved during 41% of submucous plexus (a endotoxemia.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 405 NPY (neuropeptide Y) Ruscica M, et al.

3) Cancer by specific drugs with either agonist or antagonist Besides these physiological events, in the context of the activity or by radiolabeled compounds (the universal ligand 125 I-labeled PYY, the selective Y1-R 125 I-labeled biology of cancer progression, recent evidence has 125 extended the oncological relevance of NPY to [Leu 31 , Pro 34 ]-PYY and the selective Y2-R I-labeled endocrine-related cancers (see below). PYY 3-36 ); (b) in situ production of NPY. Tumor- produced NPY may act locally in a paracrine fashion Homology and/or reach the bloodstream and generate systemic The alignment of mature tetrapod NPY sequences effects. The measurement of NPY plasma levels may reveals constant positions (91% identity) and only 4 have the value of a diagnostic marker; (c) a positions are allowed to vary. In particular, amphi- combination of expression of NPY-Rs and local bians sequences display an Arg-Lys replacement at production of NPY by the tumor. NPY effects may position 19, porcine and bovine a Lys-Met replace- result in either promotion or suppression of cancer ment at position 17, ovine an Asp-Glu replacement at growth through paracrine mechanisms. position 10, and chicken an Asn-Ser replacement at NPY is then involved in specific steps of cancer position 7. On the contrary, identical sequences of progression, like cell proliferation, matrix invasion, human mature NPY have been found in monkey, rat, metastatization, and angiogenesis. Proliferative or anti- rabbit, guinea pig, and alligator. proliferative effects of NPY have been shown in different cell systems and appear mediated mainly Mutations through the Y1-R, which has been involved in the Note control of the proliferation of vascular smooth muscle and endothelial cells and injured glial cells. NPY has So far, in the literature there are many evidence which also been found to modulate the proliferation of hepatic connect NPY and NPY receptors to a condition or stellate cells and neural crest-derived tumors. disease, like hyperlipidemia, enhanced rate of atherosclerosis, increased body mass index (BMI), NPY significantly enhances also angiogenesis in higher birth weigh, as well as alcohol dependence, various ex vivo and in vivo models of angiogenesis, depression and schizophrenia. To date there are such as aortic sprouting, Matrigel plug assay, and reported only two non-synonymous sequence variants ischemic hind limb revascularization. NPY-driven in the signal sequence part of NPY gene (p.L7P; 7th aa angiogenesis has been proposed to be bimodal at least -12 -8 of the preproNPY, leucine, is changed to proline; in vitro (at low and high concentration, 10 and 10 rs16139) and c.64C>A (p.L22M; 22nd aa of the M, respectively) and through oligomerization of Y1-, preproNPY, leucine, is changed to methionine; rs5571). Y2-, and Y5-R. These data might explain the in vivo The single nucleotide polymorphism (SNP) L7P was NPY-induced angio-genesis both in tissues that contain originally discovered in 1998 in Finnish and Dutch low NPY concentrations, such as the populations. It varies from 6% to 15% in the Caucasian nonsympathetically innervated aorta or growing organs populations and it seems to be totally absent or lacking a mature NPY system and in tissues containing extremely low in oriental populations. L7P substitution high NPY concentrations, such as the heart, mature is located in the signal peptide part of preproNPY and it vessels or muscles during high sympathetic activity is due to a single base substitution c.20T>C. induced by stress, ischemia, or injury. Nevertheless, by using an in vitro assay to test the multistage process of Implicated in metastasis (cell attachment to extra-cellular matrix (ECM), enzymatic degradation of ECM, and migration Endocrine-related cancers to fibronectin) it has been demonstrated that NPY Note affects tumor matrix invasion. In particular, it inhibits Endocrine-related cancers are malignancies depending the invasion of murine Colon 26-L5 adenocarcinoma at least in part upon the trophic effect of specific cells through a reconstituted basement membrane, hormones, and include breast, ovarian and prostate whereas on both the DU145 and PC-3 human androgen cancers, and to endocrine (pituitary tumors, independent prostate cancer cells it was not. In none of adrenocortical lesions) and neuroendocrine (pheo- the tumor cell lines considered, as neither on LNCaP chromocytoma, neuroblastoma, gastroentero-pancreatic human androgen dependent prostate cancer cells NPY tumors-PETs) tumors. enhanced the haptotactic migration. Oncogenesis Atherosclerosis The involvement of NPY and NPY receptors (NPY-Rs) Note in the progression of endocrine-related cancers may be In 1998 it has been reported that the P7 allele (p.P7) in associated with: (a) expression of NPY-Rs in the the NPY signalling peptide could be strongest genetic context of the tumor, thus becoming a target of factor influencing serum cholesterol and low density extratumorally secreted NPY proximal to the tumor. In lipo-protein (LDL) levels in obese subject. Two this context, the presence of NPY-Rs may be targeted independent studies showed that: a) men with P7

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 406 NPY (neuropeptide Y) Ruscica M, et al.

substitution had a four-year increase in the mean and catecholamine concentrations. J Endocrinol. 1988 maximal common carotid intima-media thickness Mar;116(3):421-6 (IMT), b) increased carotid IMT in type 2 diabetes Sundler F, Grunditz T, Håkanson R, Uddman R. Innervation of patients, c) obese subject with BMI > 30 Kg/m2 had the thyroid. A study of the rat using retrograde tracing and increased serum total cholesterol and LDL cholesterol. immunocytochemistry. Acta Histochem Suppl. 1989;37:191-8 On the contrary, in 443 children with a family history Schwartz TW, Fuhlendorff J, Kjems LL, Kristensen MS, of cardiovas-cular disease (CVD) the Leu7Pro Vervelde M, O'Hare M, Krstenansky JL, Bjørnholm B. Signal epitopes in the three-dimensional structure of neuropeptide Y. polymorphism was not associated with serum lipid Interaction with Y1, Y2, and pancreatic polypeptide receptors. values after adjustment for body weight in boys or Ann N Y Acad Sci. 1990;611:35-47 girls, but may be associated with dietary response to Ullman B, Franco-Cereceda A, Hulting J, Lundberg JM, Sollevi LDL-cholesterol concentration in overweight boys with A. Elevation of plasma neuropeptide Y-like immunoreactivity a family history of early-onset CVD. and noradrenaline during myocardial ischaemia in man. J Obesity Intern Med. 1990 Dec;228(6):583-9 Dumont Y, Martel JC, Fournier A, St-Pierre S, Quirion R. Note Neuropeptide Y and neuropeptide Y receptor subtypes in brain Although the role of NPY in the regulation of human and peripheral tissues. Prog Neurobiol. 1992;38(2):125-67 feeding is not fully clear and there are several negative Wulff BS, Catipovic B, Okamoto H, Gether U, Schwartz TW, studies, in 2005 a study conducted on Dutch population Johansen TE. Efficient amidation of C-peptide deleted NPY of non-obese subjects, highlighted that both men and precursors by non-endocrine cells is affected by the presence women, carried the L7P substitution, had higher mean of Lys-Arg at the C-terminus. Mol Cell Endocrinol. 1993 Feb;91(1-2):135-41 BMI values. On the other hand, one conflicting study associated the p.L7P with lower BMI in premenopausal Grouzmann E, Werffeli-George P, Fathi M, Burnier M, Waeber B, Waeber G. Angiotensin-II mediates norepinephrine and women. neuropeptide-Y secretion in a human pheochromocytoma. J Diabetes Clin Endocrinol Metab. 1994 Dec;79(6):1852-6 Note Nichols K, Staines W, Krantis A. 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J Comp Neurol. 1985 Nov 8;241(2):138-53 hypothalamic gene expression of neuropeptide Y, galanin, proopiomelanocortin, and adipocyte leptin gene expression Kerkérian L, Pelletier G. Effects of monosodium L-glutamate and secretion: effects of food restriction. Endocrinology. 1999 administration on neuropeptide Y-containing neurons in the rat Jun;140(6):2868-75 hypothalamus. Brain Res. 1986 Mar 26;369(1-2):388-90 Caberlotto L, Fuxe K, Hurd YL. Characterization of NPY Watson JD, Sury MR, Corder R, Carson R, Bouloux PM, Lowry mRNA-expressing cells in the human brain: co-localization with PJ, Besser GM, Hinds CJ. Plasma levels of neuropeptide Y2 but not Y1 mRNA in the cerebral cortex, hippocampus, tyrosine Y (NPY) are increased in human sepsis but are amygdala, and striatum. J Chem Neuroanat. 2000 Dec;20(3- unchanged during canine endotoxin shock despite raised 4):327-37

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Cerdá-Reverter JM, Larhammar D. Neuropeptide Y family of Gehlert DR. Introduction to the reviews on neuropeptide Y. peptides: structure, anatomical expression, function, and Neuropeptides. 2004 Aug;38(4):135-40 molecular evolution. Biochem Cell Biol. 2000;78(3):371-92 Pettersson-Fernholm K, Karvonen MK, Kallio J, Forsblom CM, Helisalmi S, Valve R, Karvonen MK, Hiltunen M, Pirskanen M, Koulu M, Pesonen U, Fagerudd JA, Groop PH. Leucine 7 to Mannermaa A, Koulu M, Pesonen U, Uusitupa M, Soininen H. proline 7 polymorphism in the preproneuropeptide Y is The leucine (7)-to-proline (7) polymorphism in the signal associated with proteinuria, coronary heart disease, and peptide of neuropeptide Y is not associated with Alzheimer's glycemic control in type 1 diabetic patients. Diabetes Care. disease or the link apolipoprotein E. Neurosci Lett. 2000 Jun 2004 Feb;27(2):503-9 16;287(1):25-8 Kitlinska J, Abe K, Kuo L, Pons J, Yu M, Li L, Tilan J, Everhart Kallio J, Pesonen U, Karvonen MK, Kojima M, Hosoda H, L, Lee EW, Zukowska Z, Toretsky JA. Differential effects of Kangawa K, Koulu M. Enhanced exercise-induced GH neuropeptide Y on the growth and vascularization of neural secretion in subjects with Pro7 substitution in the prepro-NPY. crest-derived tumors. Cancer Res. 2005 Mar 1;65(5):1719-28 J Clin Endocrinol Metab. 2001 Nov;86(11):5348-52 Nordman S, Ding B, Ostenson CG, Kärvestedt L, Brismar K, Karvonen MK, Valkonen VP, Lakka TA, Salonen R, Koulu M, Efendic S, Gu HF. Leu7Pro polymorphism in the neuropeptide Pesonen U, Tuomainen TP, Kauhanen J, Nyyssönen K, Lakka Y (NPY) gene is associated with impaired glucose tolerance HM, Uusitupa MI, Salonen JT. Leucine7 to proline7 and type 2 diabetes in Swedish men. Exp Clin Endocrinol polymorphism in the preproneuropeptide Y is associated with Diabetes. 2005 May;113(5):282-7 the progression of carotid atherosclerosis, blood pressure and serum lipids in Finnish men. Atherosclerosis. 2001 Ruscica M, Dozio E, Boghossian S, Bovo G, Martos Riaño V, Nov;159(1):145-51 Motta M, Magni P. Activation of the Y1 receptor by neuropeptide Y regulates the growth of prostate cancer cells. Nagakawa O, Ogasawara M, Murata J, Fuse H, Saiki I. Effect Endocrinology. 2006 Mar;147(3):1466-73 of prostatic neuropeptides on migration of prostate cancer cell lines. Int J Urol. 2001 Feb;8(2):65-70 van Rossum CT, Pijl H, Adan RA, Hoebee B, Seidell JC. Polymorphisms in the NPY and AGRP genes and body fatness Cavadas C, Ribeiro CA, Cotrim M, Mosimann F, Brunner HR, in Dutch adults. Int J Obes (Lond). 2006 Oct;30(10):1522-8 Grouzmann E. Catecholamine and neuropeptide Y secretion from human adrenal chromaffin cells: effect of nicotine and Ruscica M, Dozio E, Motta M, Magni P. Relevance of the KCl. Ann N Y Acad Sci. 2002 Oct;971:332-4 neuropeptide Y system in the biology of cancer progression. Curr Top Med Chem. 2007;7(17):1682-91 Mattevi VS, Zembrzuski VM, Hutz MH. Association analysis of genes involved in the leptin-signaling pathway with obesity in Ruscica M, Dozio E, Motta M, Magni P. Role of neuropeptide Y Brazil. Int J Obes Relat Metab Disord. 2002 Sep;26(9):1179-85 and its receptors in the progression of endocrine-related cancer. Peptides. 2007 Feb;28(2):426-34 Lee EW, Grant DS, Movafagh S, Zukowska Z. Impaired angiogenesis in neuropeptide Y (NPY)-Y2 receptor knockout Ukkola O, Kesäniemi YA. Leu7Pro polymorphism of mice. Peptides. 2003 Jan;24(1):99-106 PreproNPY associated with an increased risk for type II diabetes in middle-aged subjects. Eur J Clin Nutr. 2007 Pedrazzini T, Pralong F, Grouzmann E. Neuropeptide Y: the Sep;61(9):1102-5 universal soldier. Cell Mol Life Sci. 2003 Feb;60(2):350-77 Salminen M, Lehtimäki T, Fan YM, Vahlberg T, Kivelä SL. Ubink R, Calza L, Hökfelt T. 'Neuro'-peptides in glia: focus on Leucine 7 to proline 7 polymorphism in the neuropeptide Y NPY and galanin. Trends Neurosci. 2003 Nov;26(11):604-9 gene and changes in serum lipids during a family-based counselling intervention among school-aged children with a von Eggelkraut-Gottanka R, Machova Z, Grouzmann E, family history of CVD. Public Health Nutr. 2008 Beck-Sickinger AG. Semisynthesis and characterization of the Nov;11(11):1156-62 first analogues of pro-neuropeptide y. Chembiochem. 2003 May 9;4(5):425-33 This article should be referenced as such: Ruscica M, Dozio E, Magni P. NPY (neuropeptide Y). Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4):404-408.

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Gene Section Mini Review

ORAOV1 (oral cancer overexpressed 1) Lu Jiang, Jinsheng Yu, Qianming Chen State Key Laboratory of Oral Diseases, West China College of Stomatology, Sichuan University, Chengdu, China (LJ), School of public health, Center for Molecular Biology of Oral Diseases, University of Illinois at Chicago, Chicago, IL, USA (JY), School of public health, Center for Molecular Biology of Oral Diseases, University of Illinois at Chicago, Chicago, IL, USA (QC)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/ORAOV1ID41611ch11q13.html DOI: 10.4267/2042/44738 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

degree of oral squamous cell carcinoma has been Identity observed in a Chinese patient cohort. Other names: TAOS1 HGNC (Hugo): ORAOV1 Protein Location: 11q13.2 Note So far, no study describes the location of ORAOV1 DNA/RNA protein in cells or tissues. Note Function The ORAOV1 gene spans 9.75 Kb genomic DNA and Apoptosis, but whether it belongs to extrinsic or intrisic consists of 5 known exons. The coding sequence of pathway is yet unknown. ORAOV1 is 414 nucleotides long. Transcription Implicated in So far, there is one reported transcript variant of Oral squamous cell carcinoma ORAOV1 caused by alternative exon utilization. The Note production of transcript variant 1, which is ORAOV1 gene may be a candidate biomarker for named as ORAOV1-A, is produced by alternative diagnosis and prognosis in oral squamous cell splicing of exon 3. An in-frame stop codon in exon 4 carcinoma (OSCC). This finding was made based on a indicates that the ORAOV1 protein isoform 1 may be study using PCR to measure ORAOV1 in 15 healthy interrupted at this position. Interestingly, an association oral mucosa tissue samples, 30 OLK tissues samples, between the expression level of this ORAOV1 gene and 33 OSCC tissue samples. ORAOV1-A and the differentiation

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 409 ORAOV1 (oral cancer overexpressed 1) Jiang L, et al.

Figure 1 : ORAOV1 gene consists of five exons and spans 9.75 kb of genomic DNA. Figure 2 : (A) The schematic structures of the ORAOV1 gene and its two transcript variants. ORAOV1 has five exons. The two ORAOV1 splice variants consist of different exon combinations that produce different protein-coding regions. (B) Comparison of ORAOV1 with ORAOV1-A nucleotide sequence. White boxes show amino acid differences.

The authors found that ORAOV1 amplification was The latter is most likely due to a suppression of VEGF. significant-ly associated with larger tumor size, The study of the novel splice variant of ORAOV1, presence of lymph node metastasis, poor histological ORAOV1-A, showed that there was an inverse differen-tiation and advanced clinical stage. Therefore, correlation between the expression frequency of they suggested the potential roles of ORAOV1 in oral ORAOV1-A and the degree of differentiation in OSCC. carcinogenesis. Further studies about the biological This result suggested that ORAOV1-A may play a functional role of ORAOV1 in OSCC tumori-genesis functional role in the tumorigenesis of OSCC, and the showed that ORAOV1 plays pivotal roles in the growth ORAOV1-A expression may serve as an adjunctive and angiogenesis of OSCC. The authors performed a prognostic indicator for patients with OSCC. loss-of-function study by using small interfering RNA (siRNA) against ORAOV1 in two OSCC cell lines. Esophageal squamous cell carcinoma They found that the OSCC cells with reduced Note ORAOV1 showed retarded cell growth in vitro and ORAOV1 is supposed to serve as a new marker for displayed inhibition in both tumor growth and tumor predicting the malignancy of esophageal SCC. This angiogenesis in vivo. Further analyses reveal that the finding was made based on a study using quantitative retarded cell growth is associated with an increase in RT-PCR to examine the ORAOV1 gene expression apoptosis involving the activation of caspase 3- level in 38 esophageal SCCs. The authors found that dependent pathway, and a cell cycle arrest at the S- the ORAOV1 overexpression correlated with the phase with a downregulation of cyclin A, cyclin B1, clinicopathological features of esophageal SCCs, which and CDC2. The suppressed tumor growth in vivo may revealed a significant difference in lymph node be attributed to synergistic effect between inhibition in metastasis and a trend towards advanced TNM stages. cell growth and suppression of tumor angio-genesis.

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multiple genes flanked by segmental duplications. Hum Genet. References 2007 Apr;121(2):187-201 Huang X, Gollin SM, Raja S, Godfrey TE. High-resolution Salmon Hillbertz NH, Isaksson M, Karlsson EK, Hellmén E, mapping of the 11q13 amplicon and identification of a gene, Pielberg GR, Savolainen P, Wade CM, von Euler H, Gustafson TAOS1, that is amplified and overexpressed in oral cancer U, Hedhammar A, Nilsson M, Lindblad-Toh K, Andersson L, cells. Proc Natl Acad Sci U S A. 2002 Aug 20;99(17):11369-74 Andersson G. Duplication of FGF3, FGF4, FGF19 and ORAOV1 causes hair ridge and predisposition to dermoid Katoh M, Katoh M. FLJ10261 gene, located within the CCND1- sinus in Ridgeback dogs. Nat Genet. 2007 Nov;39(11):1318-20 EMS1 locus on human chromosome 11q13, encodes the eight- transmembrane protein homologous to C12orf3, C11orf25 and Xia J, Chen Q, Li B, Zeng X. Amplifications of TAOS1 and FLJ34272 gene products. Int J Oncol. 2003 Jun;22(6):1375-81 EMS1 genes in oral carcinogenesis: association with clinicopathological features. Oral Oncol. 2007 May;43(5):508- Katoh M, Katoh M. Evolutionary conservation of CCND1- 14 ORAOV1-FGF19-FGF4 locus from zebrafish to human. Int J Mol Med. 2003 Jul;12(1):45-50 Jiang L, Zeng X, Yang H, Wang Z, Shen J, Bai J, Zhang Y, Gao F, Zhou M, Chen Q. Oral cancer overexpressed 1 Jin C, Jin Y, Gisselsson D, Wennerberg J, Wah TS, Strömbäck (ORAOV1): a regulator for the cell growth and tumor B, Kwong YL, Mertens F. Molecular cytogenetic angiogenesis in oral squamous cell carcinoma. Int J Cancer. characterization of the 11q13 amplicon in head and neck 2008 Oct 15;123(8):1779-86 squamous cell carcinoma. Cytogenet Genome Res. 2006;115(2):99-106 Jiang L, Yang HS, Wang Z, Zhou Y, Zhou M, Zeng X, Chen QM. ORAOV1-A correlates with poor differentiation in oral Komatsu Y, Hibi K, Kodera Y, Akiyama S, Ito K, Nakao A. cancer. J Dent Res. 2009 May;88(5):433-8 TAOS1, a novel marker for advanced esophageal squamous cell carcinoma. Anticancer Res. 2006 May-Jun;26(3A):2029-32 This article should be referenced as such: Gibcus JH, Kok K, Menkema L, Hermsen MA, Mastik M, Kluin Jiang L, Yu J, Chen Q. ORAOV1 (oral cancer overexpressed PM, van der Wal JE, Schuuring E. High-resolution mapping 1). Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4):409- identifies a commonly amplified 11q13.3 region containing 411.

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Gene Section Review

PAX8 (paired box 8) Dario de Biase, Luca Morandi, Giovanni Tallini Bologna University School of Medicine, Anatomia Patologica, Ospedale Bellaria, Via Altura 3, 40139 Bologna, Italy (DdB, LM, GT)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/PAX8ID382ch2q13.html DOI: 10.4267/2042/44739 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Expression Pax8 is expressed in embryonal human tissues, in HGNC (Hugo): PAX8 particular in the developing thyroid gland, kidney, Location: 2q13 Müllerian structures, and nervous system (e.g. otic placode), and in the human placenta. During thyroid DNA/RNA development PAX8 is expressed in the thyroid anlage. During kidney development it is expressed in the S- shaped body and in the early proximal tube, but is absent in the uretic bud and condensing mesenchyme. In the adult Pax8 is expressed in the thyroid gland and kidney but it is absent in most other developed organs. Structure of PAX8 gene. Pax8 expression has been described in several tumor Description types including thyroid and ovarian carcinomas and 62924 bp, 11 exons, cDNA 4065 bp. Wilms' tumor. Transcription Localisation Five different isoforms due to alternative splicing. Cell nucleus. Protein Function The paired box (PAX) genes code for a family of Description transcription factors containing a paired box domain, an octapeptide, and a paired-type homeo-domain. PAX Pax8 is a transcription factor. The molecular weight of proteins are essential for the forma-tion of several the unprocessed precursor is ~48 kD. Five different tissues from all germ layers in the mammalian embryo. isoforms have been described (a-e): Pax8 is a member of this family with a crucial role in -Isoform a (450 aa, 4065 bp) the morphogenesis of the thyroid gland. It also has an -Isoform b (387 aa, 3876 bp, lack exon 8; mass of ~42 important organogenetic role for the kidney, Müllerian kD) system and inner ear. -Isoform c (398 aa, 3986 bp, lack exons 7, 8; mass of In the thyroid, Pax8 is a master gene that regulates ~43 kD) maintenance of the differentiated thyroid follicular cell -Isoform d (321 aa, 3755 bp, lack exon 8; mass of ~35 phenotype. As such it controls and activates the kD) transcription and thyroid specific expression of the main proteins responsible for the functional activity of -Isoform e (287 aa, 3653 bp, lack exons 8, 9, 10; mass follicular cells such as TG (thyroglobulin), TPO of ~31 kD)

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(thyroperoxidase) and NIS (sodium/iodide symporter). characterization of the 2q13 and 3p25 translocation In the developing Kidney PAX8 is important for renal breakpoint regions revealed fusion between exons 1 to vescicle formation and regulates the expression of the 7, 1 to 8, or 1 to 9 of PAX8 (2q13) and exons 1 to 6 of WT1 gene. PPARgamma1 (3p25). Homology Abnormal protein PAX8/PPARgamma1 (PPFP) PAX family members (especially PAX2). Mutations Note Germ line PAX8 mutations are a cause of congenital hypothyroidism non-goitrous type 2 (CHNG2). PAX8 rearrangement and fusion with PPARgamma1 are associated with the development of thyroid tumors of follicular cell derivation. Germinal -R31H (Sporadic, Hypoplasia, Hypothyroid) -Q40P (Sporadic, Hypoplasia, Hypothyroid) -C57Y (Familial, Hypoplasia, Hypothyroid) -L62R (Familial, Hypoplasia, Cystic rudiment, Hypothyroid) -R108TER (Sporadic, Ectopia with hypoplasia, Elevated TSH, elevated TG) -R31C -R52P -S54G -S48F -T225M -F329L (Polymorphism) -c.989_992delACCC

Somatic Oncogenesis t(2;3)(q13;p25): PAX8/PPARgamma Fusion Gene Translocations or inversions can give rise to the activation of an oncogene through its positioning near a Implicated in strong promoter or its fusion with another gene, Thyroid tumors endowing the fused transcript with tumori-genic properties. Note PAX8 is a transcription factor with a key role in the PAX8/PPARgamma1 is an oncoprotein resulting from maintenance of a differentiated phenotype in thyroid fusion of the DNA-binding domains of PAX8 to follicular cells. PPARgamma is a ligand dependent domains A to F of the peroxisome proliferator- nuclear transcription factor highly expressed in adipose activated receptor gamma-1 (PPARgamma1). It tissue. involves a chromosome 3p25 and 2q13 transloca-tion, creating a fusion gene. This encompasses the promoter The PAX8 promoter, which is active in thyroid and the proximal coding portion of PAX8 gene and follicular cells, drives the expression of PAX8/ most of the coding sequence of the PPARgamma1 PPARgamma1 fusion protein (PPFP). Since no point gene. PPARgamma1 maps to 3p25, which is a mutations in the PPARgamma1 gene have been found breakpoint hot spot region for thyroid tumors of in thyroid carcinomas and cell lines, it is speculated follicular cell origin (follicular carcino-mas and that PPARgamma1 activation resulting from the adenomas). rearrangement plays a direct oncogenetic role. Reduced expression of normal PAX8 protein may also Cytogenetics contribute to tumor development. t(2;3)(q13;p25). Fine mapping and molecular

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PD: Paired Homeobox (DNA Binding Domain); HD: Partial Homeobox; AD: Activation Domain; DBD: DNA Binding Domain; LB: Ligand Binding

In fact, PPFP oncogenic effects could relate to Papillary thyroid carcinoma, follicular constitutive overexpression of the full length variant PPARgamma1 domain, interference with wild-type PPARgamma1 or PAX8 function, novel intrinsic Note properties of PPFP, or a combination of the above. PAX8/PPARgamma1 has been identified in some PPARgamma1 is thought to be the principal target of a papillary thyroid carcinomas, follicular variant (up to class of antidiabetic agents (thiazolidinediones) and 30% in some series). PPFP inhibits thiazolidine-dione-induced gene Disease transactivation by wild-type PPARgamma1. A malignant epithelial tumor showing evidence of PAX8/PPARgamma1 has been identified in thyroid follicular cell differentiation and characterized by tumors of follicular cell deriva-tion characterized by a distinctive nuclear features (enlargement, oval shape, well developed follicular pattern of growth. These elongation and overlapping). Follicular variant tumors are usually follicular carcinomas but may also papillary carcinoma is composed of folli-cular be follicular variant papillary carcinomas or follicular structures without well developed papillae. adenomas. It has therefore been suggested that PAX8/ Thyroid follicular adenoma PPARgamma1 represents an early event in follicular cell tumorigenesis. Diagnosis of thyroid malignancies Note with a follicular growth pattern is primarily based on PAX8/PPARgamma1 has been identified in ~5-10% of the identification of capsular or vascular invasion, thyroid follicular adenoma. which can only be assessed by histopathological Disease examination of the surgically removed specimen. As a A benign, encapsulated tumor of the thyroid showing consequence, many individuals diagnosed with a evidence of follicular cell differentiation. follicular-patterned thyroid neoplasm undergo surgery. Prognosis Since PAX8/ PPARgamma1 is associated with a diagnosis of carcinoma, identification of the Thyroid follicular adenomas are benign tumors. rearrangement may prove a useful tool for molecular Wilms' tumor diagnosis. Note Thyroid follicular carcinoma PAX8 is expressed in Wilms' tumor and it is potentially involved in its induction. Pax-8 gene product resides Note upstream of wt1 in a common regulatory pathway. Two PAX8/PPARgamma1 has been identified in thyroid PAX8 isoforms, genera-ted by alternative splicing at tumors of follicular cell derivation characterized by a the C-terminus, were found to be capable of activating well developed follicular pattern of growth. These wt1 expression. PAX8 function during mesenchymal- tumors are usually follicular carcinomas but may also epithelial cell transition in renal development is to be follicular variant papillary carcinomas or follicular induce wt1 gene expression. adenomas. Disease Disease Wilms' tumor, or nephroblastoma, is a malignant Thyroid follicular carcinoma is a malignant epithelial embryonal neoplasm of the kidney derived from tumor showing evidence of follicular cell nephrogenic blastemal cells that replicate the histology differentiation and lacking the diagnostic nuclear of the developing kidney. The tumor often shows features of papillary carcinoma. divergent differentiation patterns. It is characterized by Prognosis abortive tubules and glomeruli, surrounded by a PAX8/PPARgamma1 is detected in ~30-40% of spindled cell stroma. The stroma may include striated thyroid follicular carcinomas. Follicular carcinomas muscle, cartilage, bone, fat tissue. The genes with PAX8/PPARgamma1 are angioinvasive and may predisposing to Wilms' tumor are WT1 and WT2. be aggressive. Prognosis Wilms' tumor is aggressive but potentially curable.

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Ovarian carcinoma but if caused by organification defects is often recessively inherited. Note Aberrant transcriptional expression of PAX8 has been The mutant proteins have markedly reduced DNA reported in epithelial ovarian cancer. binding with subsequent loss of transcriptional activation function. The mutations are thought to Disease disrupt the pronounced gain of alpha helical PAX8 Malignant tumors of the ovary derived from the ovarian content that follows the interaction of PAX8 with surface epithelium or its derivatives. DNA: they impair the unstructured to structured Acute myeloid leukaemia transition that occurs during DNA recognition (loss of Note "induced fit"). As a result of the mutations PAX8 protein cannot perform its role in activating PAX8 (with PAX2) may be a candidate for the transcription of its target-genes, such as TG, TPO and upregulation of WT1 in a proportion of Acute myeloid NIS. leukaemias. Marked phenotypic variability has been found within Disease affected families, suggesting variable penetrance and Acute myeloid leukemia is a neoplasm of myeloid expressivity of PAX8 gene defects. blasts in bone marrow, blood or other tissue. Some PAX8 mutations cause congenital hypothyro- Congenital hypothyroidism non- idism, while others mildly reduce thyroid hormone goitrous type 2 (CHNG2) levels or have no detectable effect. Accordingly, Note thyroid glands of patients with PAX8 mutations are Several PAX8 mutations have been identified often small and hypoplastic, sometimes completely absent (athyreosis), suppor-ting the concept that PAX8 located in the conserved paired domain of PAX8. mutations disrupt the normal growth and survival of Disease thyroid cells during embryonic development. The Congenital hypothyroidism non-goitrous type 2 reduced thyroid parenchymal mass may be unable to (CHNG2) is a congenital form of hypothyroidism due produce the required amounts of thyroid hormone. to thyroid dysgenesis (athyreotic hypothyro-idism), However, normal thyroid glands have also been while congenital hypothyroidism non-goitrous type 1 reported in patients carrying PAX8 mutations. A small (CHNG1) is due to resistance to thyroid-stimulating deletion (c.989_992delACCC) in exon 7 causing a hormone (TSH). frame-shift with premature stop codon after codon 277 Primary congenital hypothyroidism is usually sporadic has been described in a patient with congenital hypothyroidism and thyroid hypoplasia. Breakpoints

PAX8/PPARgamma1 breakpoints and chimeric mRNA.

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oncogene in human thyroid carcinoma [corrected]. Science. References 2000 Aug 25;289(5483):1357-60 Plachov D, Chowdhury K, Walther C, Simon D, Guenet JL, Pasca di Magliano M, Di Lauro R, Zannini M. Pax8 has a key Gruss P. Pax8, a murine paired box gene expressed in the role in thyroid cell differentiation. Proc Natl Acad Sci U S A. developing excretory system and thyroid gland. Development. 2000 Nov 21;97(24):13144-9 1990 Oct;110(2):643-51 Congdon T, Nguyen LQ, Nogueira CR, Habiby RL, Medeiros- Poleev A, Fickenscher H, Mundlos S, Winterpacht A, Zabel B, Neto G, Kopp P. A novel mutation (Q40P) in PAX8 associated Fidler A, Gruss P, Plachov D. PAX8, a human paired box with congenital hypothyroidism and thyroid hypoplasia: gene: isolation and expression in developing thyroid, kidney evidence for phenotypic variability in mother and child. J Clin and Wilms' tumors. Development. 1992 Nov;116(3):611-23 Endocrinol Metab. 2001 Aug;86(8):3962-7 Zannini M, Francis-Lang H, Plachov D, Di Lauro R. Pax-8, a Vilain C, Rydlewski C, Duprez L, Heinrichs C, Abramowicz M, paired domain-containing protein, binds to a sequence Malvaux P, Renneboog B, Parma J, Costagliola S, Vassart G. overlapping the recognition site of a homeodomain and Autosomal dominant transmission of congenital thyroid activates transcription from two thyroid-specific promoters. Mol hypoplasia due to loss-of-function mutation of PAX8. J Clin Cell Biol. 1992 Sep;12(9):4230-41 Endocrinol Metab. 2001 Jan;86(1):234-8 Kozmik Z, Kurzbauer R, Dörfler P, Busslinger M. Alternative Fan Y, Newman T, Linardopoulou E, Trask BJ. Gene content splicing of Pax-8 gene transcripts is developmentally regulated and function of the ancestral chromosome fusion site in human and generates isoforms with different transactivation chromosome 2q13-2q14.1 and paralogous regions. Genome properties. Mol Cell Biol. 1993 Oct;13(10):6024-35 Res. 2002 Nov;12(11):1663-72 Stapleton P, Weith A, Urbánek P, Kozmik Z, Busslinger M. Martelli ML, Iuliano R, Le Pera I, Sama' I, Monaco C, Chromosomal localization of seven PAX genes and cloning of Cammarota S, Kroll T, Chiariotti L, Santoro M, Fusco A. a novel family member, PAX-9. Nat Genet. 1993 Apr;3(4):292- Inhibitory effects of peroxisome poliferator-activated receptor 8 gamma on thyroid carcinoma cell growth. J Clin Endocrinol Metab. 2002 Oct;87(10):4728-35 Eccles MR, Yun K, Reeve AE, Fidler AE. Comparative in situ hybridization analysis of PAX2, PAX8, and WT1 gene Ambroziak M, Pachucki J, Chojnowski K, Wiechno W, Nauman transcription in human fetal kidney and Wilms' tumors. Am J J, Nauman A. Pax-8 expression correlates with type II 5' Pathol. 1995 Jan;146(1):40-5 deiodinase expression in thyroids from patients with Graves' disease. Thyroid. 2003 Feb;13(2):141-8 Dehbi M, Pelletier J. PAX8-mediated activation of the wt1 tumor suppressor gene. EMBO J. 1996 Aug 15;15(16):4297- Cheung L, Messina M, Gill A, Clarkson A, Learoyd D, 306 Delbridge L, Wentworth J, Philips J, Clifton-Bligh R, Robinson BG. Detection of the PAX8-PPAR gamma fusion oncogene in Dörfler P, Busslinger M. C-terminal activating and inhibitory both follicular thyroid carcinomas and adenomas. J Clin domains determine the transactivation potential of BSAP (Pax- Endocrinol Metab. 2003 Jan;88(1):354-7 5), Pax-2 and Pax-8. EMBO J. 1996 Apr 15;15(8):1971-82 Dwight T, Thoppe SR, Foukakis T, Lui WO, Wallin G, Höög A, Mansouri A, Hallonet M, Gruss P. Pax genes and their roles in Frisk T, Larsson C, Zedenius J. Involvement of the cell differentiation and development. Curr Opin Cell Biol. 1996 PAX8/peroxisome proliferator-activated receptor gamma Dec;8(6):851-7 rearrangement in follicular thyroid tumors. J Clin Endocrinol Kozmik Z, Czerny T, Busslinger M. Alternatively spliced Metab. 2003 Sep;88(9):4440-5 insertions in the paired domain restrict the DNA sequence Nikiforova MN, Lynch RA, Biddinger PW, Alexander EK, Dorn specificity of Pax6 and Pax8. EMBO J. 1997 Nov GW 2nd, Tallini G, Kroll TG, Nikiforov YE. RAS point mutations 17;16(22):6793-803 and PAX8-PPAR gamma rearrangement in thyroid tumors: Paschke R, Ludgate M. The thyrotropin receptor in thyroid evidence for distinct molecular pathways in thyroid follicular diseases. N Engl J Med. 1997 Dec 4;337(23):1675-81 carcinoma. J Clin Endocrinol Metab. 2003 May;88(5):2318-26 Fabbro D, Pellizzari L, Mercuri F, Tell G, Damante G. Pax-8 Siehl JM, Thiel E, Heufelder K, Snarski E, Schwartz S, protein levels regulate thyroglobulin gene expression. J Mol Mailänder V, Keilholz U. Possible regulation of Wilms' tumour Endocrinol. 1998 Dec;21(3):347-54 gene 1 (WT1) expression by the paired box genes PAX2 and PAX8 and by the haematopoietic transcription factor GATA-1 Macchia PE, Lapi P, Krude H, Pirro MT, Missero C, Chiovato in human acute myeloid leukaemias. Br J Haematol. 2003 L, Souabni A, Baserga M, Tassi V, Pinchera A, Fenzi G, Oct;123(2):235-42 Grüters A, Busslinger M, Di Lauro R. PAX8 mutations associated with congenital hypothyroidism caused by thyroid De Felice M, Di Lauro R. Thyroid development and its dysgenesis. Nat Genet. 1998 May;19(1):83-6 disorders: genetics and molecular mechanisms. Endocr Rev. 2004 Oct;25(5):722-46 Missero C, Cobellis G, De Felice M, Di Lauro R. Molecular events involved in differentiation of thyroid follicular cells. Mol de Sanctis L, Corrias A, Romagnolo D, Di Palma T, Biava A, Cell Endocrinol. 1998 May 25;140(1-2):37-43 Borgarello G, Gianino P, Silvestro L, Zannini M, Dianzani I. Familial PAX8 small deletion (c.989_992delACCC) associated Torban E, Goodyer P. What PAX genes do in the kidney. Exp with extreme phenotype variability. J Clin Endocrinol Metab. Nephrol. 1998 Jan-Feb;6(1):7-11 2004 Nov;89(11):5669-74 Ohno M, Zannini M, Levy O, Carrasco N, di Lauro R. The Gregory Powell J, Wang X, Allard BL, Sahin M, Wang XL, Hay paired-domain transcription factor Pax8 binds to the upstream ID, Hiddinga HJ, Deshpande SS, Kroll TG, Grebe SK, enhancer of the rat sodium/iodide symporter gene and Eberhardt NL, McIver B. The PAX8/PPARgamma fusion participates in both thyroid-specific and cyclic-AMP-dependent oncoprotein transforms immortalized human thyrocytes transcription. Mol Cell Biol. 1999 Mar;19(3):2051-60 through a mechanism probably involving wild-type PPARgamma inhibition. Oncogene. 2004 Apr 29;23(20):3634- Kroll TG, Sarraf P, Pecciarini L, Chen CJ, Mueller E, 41 Spiegelman BM, Fletcher JA. PAX8-PPARgamma1 fusion

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 416 PAX8 (paired box 8) de Biase D, et al.

Hans S, Liu D, Westerfield M. Pax8 and Pax2a function ret, Braf , ras and pax8 genes. Endocr Relat Cancer. 2006 synergistically in otic specification, downstream of the Foxi1 Jun;13(2):485-95 and Dlx3b transcription factors. Development. 2004 Oct;131(20):5091-102 Drieschner N, Belge G, Rippe V, Meiboom M, Loeschke S, Bullerdiek J. Evidence for a 3p25 breakpoint hot spot region in Park SM, Chatterjee VK. Genetics of congenital thyroid tumors of follicular origin. Thyroid. 2006 hypothyroidism. J Med Genet. 2005 May;42(5):379-89 Nov;16(11):1091-6 Trueba SS, Augé J, Mattei G, Etchevers H, Martinovic J, Al Taji E, Biebermann H, Límanová Z, Hníková O, Zikmund J, Czernichow P, Vekemans M, Polak M, Attié-Bitach T. PAX8, Dame C, Grüters A, Lebl J, Krude H. Screening for mutations TITF1, and FOXE1 gene expression patterns during human in transcription factors in a Czech cohort of 170 patients with development: new insights into human thyroid development congenital and early-onset hypothyroidism: identification of a and thyroid dysgenesis-associated malformations. J Clin novel PAX8 mutation in dominantly inherited early-onset non- Endocrinol Metab. 2005 Jan;90(1):455-62 autoimmune Castro P, Rebocho AP, Soares RJ, Magalhães J, Roque L, hypothyroidism. Eur J Endocrinol. 2007 May;156(5):521-9 Trovisco V, Vieira de Castro I, Cardoso-de-Oliveira M, Fonseca E, Soares P, Sobrinho-Simões M. PAX8- Bowen NJ, Logani S, Dickerson EB, Kapa LB, Akhtar M, PPARgamma rearrangement is frequently detected in the Benigno BB, McDonald JF. Emerging roles for PAX8 in ovarian follicular variant of papillary thyroid carcinoma. J Clin cancer and endosalpingeal development. Gynecol Oncol. 2007 Endocrinol Metab. 2006 Jan;91(1):213-20 Feb;104(2):331-7 Di Cristofaro J, Silvy M, Lanteaume A, Marcy M, Carayon P, This article should be referenced as such: De Micco C. Expression of tpo mRNA in thyroid tumors: quantitative PCR analysis and correlation with alterations of de Biase D, Morandi L, Tallini G. PAX8 (paired box 8). Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4):412-417.

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Gene Section Mini Review

PTGIS (prostaglandin I2 (prostacyclin) synthase) Inês Cebola, Miguel A Peinado Institute of Predictive and Personalized Medicine of Cancer (IMPPC), Badalona, Barcelona, Spain (IC, MAP)

Published in Atlas Database: May 2009 Online updated version : http://AtlasGeneticsOncology.org/Genes/PTGISID44219ch20q13.html DOI: 10.4267/2042/44740 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Transcription Two alternative splicing products; 5624bp (coding, Other names: CYP8; CYP8A1; EC 5.3.99.4; 1578bp ORF) and 313bp (non-coding). A 1.5kb MGC126858; MGC126860; PGIS; PTGI upstream sequence presents CG-rich and pyrimi-dine- HGNC (Hugo): PTGIS rich regions and contains consensus sequences for the Location: 20q13.13 recognition sites of Sp-1, AP-2, INF-gamma responsive element, GATA, NF-kB, glucocorticoid responsive DNA/RNA element and a CACCC box. It has been demonstrated that PTGIS transcription is Description regulated via Sp1 binding, which may affected by 64297 bases (chr20:47553818-47618114); 10 exons; polymorphisms in the promoter region. The CG-rich telomere to centromere orientation. region presents significant promoter activity.

Map of 20q13 chromosomal region.

Schematic diagram of PTGIS genomic region.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 418 PTGIS (prostaglandin I2 (prostacyclin) synthase) Cebola I, Peinado MA

PTGIS protein structure: secondary structure composition and features of interest.

Protein Implicated in Description Colorectal cancer PTGIS is a protein from the cytochrome P450 Note superfamily that consists of 500 amino acids and has a PTGIS promoter is silenced by through promoter molecular weight of 57kDa. Is a single-pass membrane hypermethylation in a large subset of colorectal protein with a single transmembrane domain (1-20) and carcinomas (also observed in colorectal adenomas and presents a heme-binding site at position 441. No post- in several colorectal cancer cell lines). The PTGIS translational modifications have been described for this silencing is an early event on tumor progression and protein. age and sex-independent. A PTGIS promoter VNTR Expression polymorphism has been associated with the risk of colorectal polyps, being the risk increased when both Abundant in heart, lung, skeletal muscle, ovary and alleles present less than 6 repeats. prostate. PTGIS is found underexpressed in most colorectal carcinomas and in several lung carcinomas Non-small cell lung cancer and hypertensive subjects. Disease Localisation PTGIS promoter is hypermethylated in lung cancer cell lines. The VNTR polymophism has also been At cellular level can be found in the endoplasmatic associated with gene silencing in lung cancer cells. reticulum membrane and in microsomes as a peripheral membrane protein. Breast cancer Function Disease The combination of the SNPs PTGIS(11)rs477627, Catalyses the isomerization of prostaglandin H2 to PTGIS(21)rs476496 and PTGIS(20)rs1066894, located prostacyclin (prostaglandin I2), a potent vasodilator in the 5' region and first intron, confers reduced and inhibitor of platelet aggregation. susceptibility to breast cancer. Homology Essential hypertension Pan troglodytes - Prostaglandin I2 (prostacyclin) Disease Synthase. A splicing mutation (T->C at the +2 position of the Canis lupus familiaris - Prostaglandin I2 (prostacyclin) donor site of the intron 9) alters the reading frame and Synthase. results in the production of a truncated protein with the Bos taurus - Prostaglandin I2 (prostacyclin) Synthase. heme-binding region deleted. The subjects found with Mus musculus - Prostaglandin I2 (prostacyclin) this mutation presented lower levels of prostacyclin and Synthase. hypertension. Rattus norvegicus - Prostaglandin I2 (prostacyclin) The polymorphism C1117A has been significantly Synthase. associated with hypertension, being the allele C Danio rerio - Prostaglandin I2 (prostacyclin) Synthase associated with higher risk the condition, although it like. does not change the aminoacid sequence. A polymorphism in the 5' region with a variable Mutations number of repeats of a 9-bp sequence (CCGCCAGCC) is associated with hypertension. The alleles less repeats Note possess less Sp1 sites and present lower promoter One splicing mutation has been associated with activity, being related with essential hypertension.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 419 PTGIS (prostaglandin I2 (prostacyclin) synthase) Cebola I, Peinado MA

higher pulse pressure in women and higher systolic ML, Soderlund C, Steward CA, Sulston JE, Swann M, blood pressure in older individuals. Sycamore N, Taylor R, Tee L, Thomas DW, Thorpe A, Tracey A, Tromans AC, Vaudin M, Wall M, Wallis JM, Whitehead SL, Cerebral infarction Whittaker P, Willey DL, Williams L, Williams SA, Wilming L, Wray PW, Hubbard T, Durbin RM, Bentley DR, Beck S, Rogers Disease J. The DNA sequence and comparative analysis of human Association between cerebral infarction and the PTGIS chromosome 20. Nature. 2001 Dec 20-27;414(6866):865-71 promoter VNTR alleles with less repeats. Nakayama T, Soma M, Saito S, Honye J, Yajima J, Rahmutula D, Kaneko Y, Sato M, Uwabo J, Aoi N, Kosuge K, Kunimoto M, Myocardial infarction Kanmatsuse K, Kokubun S. Association of a novel single Disease nucleotide polymorphism of the prostacyclin synthase gene with myocardial infarction. Am Heart J. 2002 May;143(5):797- A synonymous SNP at codon 373 (Arg373Arg) in exon 801 8 has been associated with risk of myocardial infarction in the japanese population. Nakayama T, Soma M, Watanabe Y, Hasimu B, Sato M, Aoi N, Kosuge K, Kanmatsuse K, Kokubun S, Marrow JD, Oates JA. Splicing mutation of the prostacyclin synthase gene in a family References associated with hypertension. Biochem Biophys Res Commun. 2002 Oct 11;297(5):1135-9 Hara S, Miyata A, Yokoyama C, Inoue H, Brugger R, Lottspeich F, Ullrich V, Tanabe T. Isolation and molecular Nana-Sinkam P, Golpon H, Keith RL, Oyer RJ, Sotto-Santiago cloning of prostacyclin synthase from bovine endothelial cells. S, Moore MD, Franklin W, Nemenoff RA, Geraci MW. J Biol Chem. 1994 Aug 5;269(31):19897-903 Prostacyclin in human non-small cell lung cancers. Chest. 2004 May;125(5 Suppl):141S Miyata A, Hara S, Yokoyama C, Inoue H, Ullrich V, Tanabe T. Molecular cloning and expression of human prostacyclin Frigola J, Muñoz M, Clark SJ, Moreno V, Capellà G, Peinado synthase. Biochem Biophys Res Commun. 1994 May MA. Hypermethylation of the prostacyclin synthase (PTGIS) 16;200(3):1728-34 promoter is a frequent event in colorectal cancer and associated with aneuploidy. Oncogene. 2005 Nov Wang LH, Chen L. Organization of the gene encoding human 10;24(49):7320-6 prostacyclin synthase. Biochem Biophys Res Commun. 1996 Sep 24;226(3):631-7 Nasrallah R, Hébert RL. Prostacyclin signaling in the kidney: implications for health and disease. Am J Physiol Renal Yokoyama C, Yabuki T, Inoue H, Tone Y, Hara S, Hatae T, Physiol. 2005 Aug;289(2):F235-46 Nagata M, Takahashi EI, Tanabe T. Human gene encoding prostacyclin synthase (PTGIS): genomic organization, Norata GD, Catapano AL. Molecular mechanisms responsible chromosomal localization, and promoter activity. Genomics. for the antiinflammatory and protective effect of HDL on the 1996 Sep 1;36(2):296-304 endothelium. Vasc Health Risk Manag. 2005;1(2):119-29 Iwai N, Katsuya T, Ishikawa K, Mannami T, Ogata J, Higaki J, Poole EM, Bigler J, Whitton J, Sibert JG, Potter JD, Ulrich CM. Ogihara T, Tanabe T, Baba S. Human prostacyclin synthase Prostacyclin synthase and arachidonate 5-lipoxygenase gene and hypertension : the Suita Study. Circulation. 1999 Nov polymorphisms and risk of colorectal polyps. Cancer Epidemiol 30;100(22):2231-6 Biomarkers Prev. 2006 Mar;15(3):502-8 Nakayama T, Soma M, Rehemudula D, Takahashi Y, Tobe H, Stearman RS, Grady MC, Nana-Sinkam P, Varella-Garcia M, Satoh M, Uwabo J, Kunimoto M, Kanmatsuse K. Association of Geraci MW. Genetic and epigenetic regulation of the human 5' upstream promoter region of prostacyclin synthase gene prostacyclin synthase promoter in lung cancer cell lines. Mol variant with cerebral infarction. Am J Hypertens. 2000 Cancer Res. 2007 Mar;5(3):295-308 Dec;13(12):1263-7 Scotto L, Narayan G, Nandula SV, Arias-Pulido H, Deloukas P, Matthews LH, Ashurst J, Burton J, Gilbert JG, Subramaniyam S, Schneider A, Kaufmann AM, Wright JD, Jones M, Stavrides G, Almeida JP, Babbage AK, Bagguley CL, Pothuri B, Mansukhani M, Murty VV. Identification of copy Bailey J, Barlow KF, Bates KN, Beard LM, Beare DM, Beasley number gain and overexpressed genes on chromosome arm OP, Bird CP, Blakey SE, Bridgeman AM, Brown AJ, Buck D, 20q by an integrative genomic approach in cervical cancer: Burrill W, Butler AP, Carder C, Carter NP, Chapman JC, potential role in progression. Genes Chromosomes Cancer. Clamp M, Clark G, Clark LN, Clark SY, Clee CM, Clegg S, 2008 Sep;47(9):755-65 Cobley VE, Collier RE, Connor R, Corby NR, Coulson A, Abraham JE, Harrington P, Driver KE, Tyrer J, Easton DF, Coville GJ, Deadman R, Dhami P, Dunn M, Ellington AG, Dunning AM, Pharoah PD. Common polymorphisms in the Frankland JA, Fraser A, French L, Garner P, Grafham DV, prostaglandin pathway genes and their association with breast Griffiths C, Griffiths MN, Gwilliam R, Hall RE, Hammond S, cancer susceptibility and survival. Clin Cancer Res. 2009 Mar Harley JL, Heath PD, Ho S, Holden JL, Howden PJ, Huckle E, 15;15(6):2181-91 Hunt AR, Hunt SE, Jekosch K, Johnson CM, Johnson D, Kay MP, Kimberley AM, King A, Knights A, Laird GK, Lawlor S, Lehvaslaiho MH, Leversha M, Lloyd C, Lloyd DM, Lovell JD, This article should be referenced as such: Marsh VL, Martin SL, McConnachie LJ, McLay K, McMurray Cebola I, Peinado MA. PTGIS (prostaglandin I2 (prostacyclin) AA, Milne S, Mistry D, Moore MJ, Mullikin JC, Nickerson T, synthase). Atlas Genet Cytogenet Oncol Haematol. 2010; Oliver K, Parker A, Patel R, Pearce TA, Peck AI, Phillimore BJ, 14(4):418-420. Prathalingam SR, Plumb RW, Ramsay H, Rice CM, Ross MT, Scott CE, Sehra HK, Shownkeen R, Sims S, Skuce CD, Smith

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Gene Section Mini Review

RASL11B (RAS-like, family 11, member B) Stefan Lorkowski Institute of Nutrition, Friedrich Schiller University Jena, Dornburger Str. 25, 07743 Jena, Germany (SL)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/RASL11BID44265ch4q12.html DOI: 10.4267/2042/44741 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity DNA/RNA Other names: MGC2827; MGC4499 Description HGNC (Hugo): RASL11B Gene: 4508 bp Location: 4q12 Chromosome: 4q12 Local order: Chr4:53,423,252-53,427,759 on the + mRNA: 1979 bp strand. Exon 1: 1-333 Note: Exon 2: 334-390 Mouse: chr5:74,591,351-74,595,502 (according to Mouse July 2007 Assembly). Exon 3: 391-467 Rat: chr14:36,392,946-36,397,168 (according to Rat Exon 4: 468-1962 November 2004 Assembly). CDS: 192-936 Zebrafish: chr20:59,670,592-59,673,835 (according to The human RASL11B gene spans about 4508 bp on Zebrafish March 2006 Assembly). chromosome 4q12 and comprises 4 exons encoding at least 2 different transcripts.

Schematic representation of human RASL11B mRNAs and genomic organization of the human RASL11B gene. The human RASL11B gene consists of 4 exons encoding a transcript with a total length of 1962 bp. One shorter variant with a length of 766 bp was found. The ATG start and TGA stop codons are located in exons 1 and 4, respectively.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 421 RASL11B (RAS-like, family 11, member B) Lorkowski S

Exons of the RASL11B gene are 333 bp (exon 1), 57 Function bp (exon 2), 77 bp (exon 3), and 1495 bp (exon 4) in Small GTPase belonging to a Ras subfamily of putative size. Sizes of introns are 618 bp (intron 1), 1153 bp tumor suppressor genes. (intron 2), and 780 bp (exon 3). All splice sites have canonical boundaries, starting the intron with 'gt' and Homology ending with 'ag'. The protein sequence of the RASL11 protein family is A polyadenylation signal in the untranslated region of highly conserved within different species and contains exon 4 is located at nucleotide position 1947. five conserved regions motives that comprise the G- Transcription domain of small GTPases (P-loop, switch 1 and 2, G4 and G5 box). In addition to the full-length RASL11B transcript, a truncated polyadenylated transcript of 766 bp was reported. Implicated in Full-length transcript: 1962 bp mRNA, 744 bp open Note reading frame. According to Stolle et al., RASL11B expression is Truncated transcript: 766 bp mRNA, 574 bp open induced during maturation of THP-1 monocytic cells reading frame. into macrophages and in coronary artery smooth muscle cells after treatment with TGF-beta1 suggesting Pseudogene that RASL11B may play a role in developmental No pseudogenes reported. processes or in pathophysiologies such as inflammation or cancer. Protein Pezeron et al. demonstrated that Rasl11b modulates function of the EGF-CFC coreceptor one-eyed-pinhead (oep) in zebrafish independently of the TGFbeta/Nodal pathway, which is crucial for germ layer formation. Down regulation of Rasl11b partially rescued endodermal and prechordal plate defects of zygotic homozygous oep zebrafish mutants. Rasl11b inhibitory action was observed only in animals with oep-deficient Domains within the human RASL11B protein. Domains backgrounds, suggesting that normal oep expression positions are indicated with vertical purple lines and intron prevents function of Rasl11b. On the other hand, positions are indicated with vertical red lines both showing the Rasl11b down regulation did not rescue exact position in the polypeptide sequence. mesendodermal defects in other Nodal pathway Description mutants. RASL11B is a 248 amino acid protein containing a characteristic RAS GTPase domain with typical References topology of a six-stranded beta-sheet surrounded by Stolle K, Schnoor M, Fuellen G, Spitzer M, Cullen P, Lorkowski five alpha-helices. The RASL11B protein has no S. Cloning, genomic organization, and tissue-specific typical prenylation signal, indicating that it is probably expression of the RASL11B gene. Biochim Biophys Acta. 2007 not anchored to cellular membranes. Jul-Aug;1769(7-8):514-24 Pézeron G, Lambert G, Dickmeis T, Strähle U, Rosa FM, Expression Mourrain P. Rasl11b knock down in zebrafish suppresses one- Expression of human RASL11B mRNA was eyed-pinhead mutant phenotype. PLoS One. 2008 Jan investigated in 37 tissues and 5 cell types. In tissues, 16;3(1):e1434 RASL11B transcript is widely expressed with highest This article should be referenced as such: levels in placenta. In cells RASL11B transcript shows Lorkowski S. RASL11B (RAS-like, family 11, member B). Atlas highest abundance in primary macrophages. Genet Cytogenet Oncol Haematol. 2010; 14(4):421-422. Localisation Cytosolic.

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Gene Section Review

TJP2 (tight junction protein 2 (zona occludens 2)) Lorenza Gonzalez-Mariscal, Erika Garay, Miguel Quiros, Rocio Tapia Center for Research and Advanced Studies (Cinvestav), Department of Physiology, Biophysics and Neuroscience, Mexico DF, 07360, Mexico (LGM, EG, MQ, RT)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Genes/TJP2ID44347ch9q21.html DOI: 10.4267/2042/44742 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Transcription Five isoforms of TJP2 have been identified: Other names: MGC26306; X104; ZO2; ZO-2 The A1 isoform ( CDS: 3570 nt); lacks exons C and B HGNC (Hugo): TJP2 The A2 isoform (CDS: 3129 nt); lacks exons C, B, 20 Location: 9q21.11 and 21. Local order: Telomeric to FXN gene (9q21.11). The A3 isoform (CDS: 2979 nt); lacks exons C, B, 20, 21, 22 and 23 and exhibits a longer exon 19 (979 vs DNA/RNA 213 bp, with a stop codon at bp 313). Description The C1 isoform (CDS: 3501 nt); lacks exon A. TJP2 gene exists as a single copy in the human The C2 isoform (CDS: 3057 nt); lacks exons A, 20 and genome; it contains 25 exons and is predicted to span 21. over approximately more than 90 Kb of the genomic DNA.

Schematic diagram of the TJP2 gene comprising 25 exons (in grey) and transcript variants. The sizes in base pairs of exons (above) and introns (below) are shown. *Indicates the position of the start codons. Exons C and B are expressed from promoter C. These exons are non-coding and translations starts from the ATG located in exon 2. Exon A is transcribed from promoter A. Isoforms C lack 23 aminoacids at the amino terminus in comparison with isoforms A. Exons 20 and 21 are alternatively spliced.

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 423 TJP2 (tight junction protein 2 (zona occludens 2)) Gonzalez-Mariscal L, et al.

Isoforms, structure and interactions of TJP2. a) Five isoforms to TJP2 have been reported. A and C isoforms are products of alternative promoter usage. A2 and C2 are consequence of alternative splicing of exons 20 and 21. Translation of A3 ends in exon 19. However in A3 exon 19 has 979 instead of 213 bp and exhibits a stop codon at bp 313. This situation makes isoforms A3 end with a sequence of 33 amino acids that are not present in any of the other TJP2 isoforms. Numbers refer to amino acids. b) TJP2 is a MAGUK protein with three PDZ domains, an SH3 module and a GK domain. Between the first two PDZ domains, a region rich in basic amino acids is found. After the GK domain in C terminus direction, an acidic region and a proline rich domain is present. The last three amino acids at the carboxyl end constitute a PDZ binding motif and the first 23 amino acid residues at the N-terminus are absent in C isoforms. Numbers refer to amino acids. c) TJP2 is a scaffold molecule that interacts with proteins that participate in cell adhesion, signaling and gene transcription. The names of these proteins and their site of interaction in TJP2 are indicated.

Northern blot analysis done with different probes for contain 1 nuclear localization signal (NLS; 106-122 aa) exons A and C reveals two TJP2 transcripts of the same and no exportation signals (NES) following the size, approximately 4.5 Kb, that respectively consensus L/M/IX 1-4L/V/IX 2-3L/V/IX 1-2L/I. correspond to TJP2A and TJP2C. Two isoforms named A and C arise from the activation Pseudogene of different promoters. In the A form the initiation codon located within a unique 5' end 189 bp sequence Not known. gives rise to a distinctive 23 aa segment, whereas in the C form the 5' end exhibits a 377 bp sequence that is not Protein translated. In isoforms C translation starts from a codon Description that corresponds to the second ATG in isoforms A. Isoform A2 is generated by alternative splicing of The full length TJP2 protein consists of 1190 amino isoform A1. It lacks amino acids 961-1108. acids corresponding to a theoretical molecular weight of 133.958 kDa. This size is 16% smaller than the Isoform A3 is generated by alternative splicing of apparent molecular mass of 160 kDa determined by isoform A1. It lacks amino acids 994-1190 and the SDS/PAGE. The high proline content (7.1%) may be sequence from amino acids 961 to 993 differs from responsible for this anomalous electrophoretic isoform A1. migration. The protein contains three PDZ, one SH3 Isoform C2 is generated by alternative splicing of and one GuK domain at its NH2 dlg-like terminal isoform C1. It lacks amino acids 961-1108. region, followed by a short COOH terminal non dlg- Expression like region that contains an acidic and a proline rich domain. The protein belongs to the MAGUK protein Normal tissue: At the mRNA level the A isoforms are family. At the carboxyl terminal end of TJP2 the type I abundant in the heart and brain whereas the C isoforms PDZ binding motif TEL is found. TJP2 is predicted to are expressed at a high level in kidney, pancreas, heart

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 424 TJP2 (tight junction protein 2 (zona occludens 2)) Gonzalez-Mariscal L, et al.

and placenta. In brain and skeletal muscle only the A isoform is detectable, but there is no tissue where Mutations TJP2C is solely present. TJP2 protein is strongly Germinal expressed in epithelial and endothelial cells. During mouse embryogenesis at the 16 cell stage, TJP2 Mutation 143T-->C, predicted to cause a valine to assembles at the apico-lateral contact site of alanine substitution (V48A) in TJP2 produces the loss blastomeres for the first time. The half-life of TJP2 of the alpha-helical structure of PDZ1 domain of TJP2. protein varies according to the degree of confluence of Somatic the culture, thus in canine kidney MDCK cells half Not known. lives correspond to 8.7 and 19.1 hrs in sparse and confluent cultures respectively. Epigenetics Localisation Microarray analysis indicates that in 70% of primary pancreatic cancers TJP2 is aberrantly methylated. Present at the cytoplasmic side of tight junctions in epithelial and endothelial cells. In proliferating cultures Implicated in and in cells under environmental stress TJP2 is also present at the nucleus. TJP2 enters the nucleus at the Pancreatic adenocarcinoma late G1 phase of the cell cycle and departs during Note mitosis. At the nucleus TJP2 exhibits a speckled In ductal pancreatic cancer cell lines BxPC3, Capan2, distribution and co-localizes with splicing factor SC-35 CFPAC1, Colo357, Hs766T, MiaPaCa-2, PL3, PL4, and the nuclear ribonucleo-protein scaffold attachment PL7, PL8, PL10, PL11 and PL11, TJP2 gene is factor SAF-B. In fibroblasts TJP2 gives a punctate methylated. Instead TJP2 gene is not methylated in pattern at the cell borders, and in cardiac muscle cells it ductal pancreatic cancer cell lines AsPC1, Capan1, is detected at the intercalated discs. Panc1, PL1, PL5, PL6, PL9, PL13 and PL14. Function Treatment with the demethylating agent 5-aza-2'- TJP2 is crucial during development as silencing deoxycytidine (5 Aza-dC) induces TJP2 expression in inhibits blastocele formation in mouse embryos and MiaPaCa (8.51 fold) and in Panc1 (1.41 fold) cells but knock out mice die shortly after implantation due to an not in AsPC1 and Hs766T. arrest in early gastrulation. Methylation analysis by digestion with restriction TJP2 silencing with siRNA delays in epithelial endonucleases reveals aberrant hypermethylation in PA monolayers the arrival of tight junction proteins to the promoter of isoform TJP2A from bases -383 to 87, cell membrane, triggers the novo formation of leaky which contains 62 CpG dinucleotides, in a variety of tight junctions and alters cell polarity, thus suggesting human primary pancreatic duct carcinomas and in the that TJP2 is critical for the correct assembly and neoplastic human pancreatic duct cell lines BxPC-3, function of tight junctions. TJP2 and TJP1 are CFPAC-1, Hs700T, Hs766T, MiaPaCa-2, Su.86.86. redundant in their role as promoters of claudins However, demethy-lation of the PA does not recover polymerization into tight junction strands. normal level of TJP2A protein expression in the neoplastic cell lines except Su.86.86. TJP2 inhibits transcription of human cyclin D1 (CD1) gene, decreases CD1 protein levels due to an increased Isoform TJP2A is specifically missing in pancreatic degradation of the protein at the proteosome, blocks adenocarcinoma samples and in human pancreatic duct cell cycle progression from G1 phase to S and in carcinoma cell lines with the exception of line PANC- consequence inhibits cell proliferation. These 1. observations favor the image of TJP2 as a tumor Disease suppressor protein. Pancreatic adenocarcinoma is a disease in which Besides the above mentioned, the nuclear function of malignant cells form in the regions of the pancreas that TJP2 remains elusive. It localizes in speckles with SC- have gland like properties. Although the pancreas has 35 and associates to lamin B1, to transcription factors exocrine and endocrine cells, about 95% of pancreatic Jun, Fos, C/EBP and Myc, to SAF-B, a chromatin cancers begin in exocrine cells and more than 90% of component involved in the assembly of transcriptosome tumors of the pancreas are ductal adenocarcinomas complexes, and enhances the nuclear localization of derived from the exocrine pancreatic ducts. Depending ARVCF, an Armadillo-repeat protein that associates on the extent of the cancer at the time of diagnosis, the with classical cadherins in adherens junctions. prognosis is generally poor with less than 5% of those diagnosed still alive five years after diagnosis. Homology Typically pancreatic cancer first metastasizes to TJP2 shares the following homology and identity (in regional lymph nodes, then to the liver and less parenthesis) with TJP1: 68% (54%); TJP3: 63% (45%); commonly to the lungs, although it can also directly PSD-95: 40% (25%) and Dlg: 40% (23%). invade surrounding visceral organs or metastasize to

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 425 TJP2 (tight junction protein 2 (zona occludens 2)) Gonzalez-Mariscal L, et al.

any surface in the abdominal cavity via peritoneal tissue nearly all breast cancers are called adenocarci- spread. nomas. Breast cancer is the most common type of Prostate adenocarcinoma cancer in women, affecting about 1 in 8 women. Note Testicular in situ carcinoma In a prostate microarray assay the promoter sequence of Note TJP2 from cancerous cell lines PC3, PC3M, PC3M- Loss of blood-testis barrier integrity and a decreased Pro4, PC3M-LN4 and LNCaP showed a significantly protein level expression of TJP2 is detected in testicular diminished hybridization in comparison to normal in situ carcinoma. In addition, ZO-2 immunoreactivity prostate cell lines RWPE-1, MLcsv40 and 267B1, became weak and diffuse at the blood testis barrier which indicates a greater methylation of the TJP2 region and spread to stain the entire lateral site of promoter in prostate cancer. However in prostate Sertoli cells as well as their cytoplasm, indicating adenocarcinoma cell lines loss of TJP2A protein is rare. altered localization. Disease Disease Prostate is a gland in the male reproductive system Testicular cancer is a disease in which malignant cells located below the bladder and in front of the rectum form in the tissues of one or both testicles. The testicles that produces seminal fluid that nourishes and are 2 egg-shaped glands located inside the scrotum that transports sperm. Most cells in the prostate gland are of produce testosterone and sperm. Germ cells within the the glandular type; therefore the adenocarci-noma is the testicles produce immature sperm that mature as they most common type of cancer to occur in the prostate. In travel through a network of tubules and tubes into the the United States prostate cancer is the most common epididymis. Almost all testicular cancers start in the type of cancer in men, affecting about one in 8. It germ cells. The testicular in situ carcinoma also known appears mainly in older men. as stage 0, is a noninvasive precursor of testicular germ Colon cancer cell tumors, which are the most common type of cancer in young men. In testicular in situ carcinoma, abnormal Note cells are found in the tiny tubules where the sperm cells In colon cancer loss of TJP2A protein is rare. begin to develop, and all tumor marker levels are Disease normal at this stage. Also called colorectal cancer or large bowel cancer, includes cancerous growths in the colon, rectum and Lung squamous carcinoma appendix. It is the third most common form of cancer Note and the second leading cause of cancer related death in Squamous cell carcinomas show a 76% decrease in the Western world. Colon cancer is thought to arise mRNA level for TJP2. from adenomatous polyps in the colon. Colon Disease adenocarcinoma accounts for 95% of cases of colon Lung cancer is a disease that forms in the tissues of the cancer. Colon cancer is the fourth most common cancer lung. In the United States it is the second most common of males and females in the United States. malignancy after prostate cancer in men and breast Breast cancer cancer in women. The two main types of lung cancer are small cell lung cancers and non-small cell lung Note cancers, which are diagnosed based on how the cells In comparison to normal cells, TJP2 is found at a much look under the microscope. One type of non-small cell lower protein level in malignant breast epithelia. In lung cancer is the squamous cell carcinoma. This cancerous breast TJP2 immunostaining becomes more cancer begins in squamous cells which are thin, flat diffuse and decreases in intensity. By quantitative PCR cells that look like fish scales. It is also called the level of TJP2 mRNA is slightly elevated in the epidermoid carcinoma. Squamous cell carcinoma is the breast tumor tissues compared to the controls, however second commonest type of lung cancer, accounting for this result is not statistically significant. 28% of all cases of lung cancer. Disease Breast cancer is a disease in which malignant cells Lung adenocarcinoma form in the tissues of the breast. Each breast has 15-20 Note sections called lobes, which have smaller sections Lung adenocarcinomas exhibit a 72% decrease in named lobules. Lobules end in dozens of tiny bulbs that the mRNA level of TJP2. can produce milk. The lobes, lobules and bulbs are Disease linked by thin tubes called ducts. The most common Lung cancer is a disease that forms in the tissues of the type of breast cancer is ductal carcinoma that begins in lung. In the United States it is the second most common the cells of the ducts. Cancer that begins in the lobes or malignancy after prostate cancer in men and breast lobules is called lobular carcinoma and is more often cancer in women. The two main types are small and found in both breasts. Because the breast is a glandular

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 426 TJP2 (tight junction protein 2 (zona occludens 2)) Gonzalez-Mariscal L, et al.

non-small cell lung cancers, which are diagnosed based Robles-Flores M, Alcántara-Hernández R, García-Sáinz JA. on how the cells look under the microscope. One type Differences in phorbol ester-induced decrease of the activity of protein kinase C isozymes in rat hepatocytes. Biochim Biophys of non-small cell lung cancer is the adenocarcinoma in Acta. 1991 Aug 13;1094(1):77-84 which the cancer begins in the cells that line the alveoli Duclos F, Rodius F, Wrogemann K, Mandel JL, Koenig M. The and make substances such as mucus. Lung Friedreich ataxia region: characterization of two novel genes adenocarcinoma is the most common kind of lung and reduction of the critical region to 300 kb. Hum Mol Genet. cancer in smokers and non-smokers and in people 1994 Jun;3(6):909-14 under age 45, accounting for about 40% of all lung Chlenski A, Ketels KV, Engeriser JL, Talamonti MS, Tsao MS, cancers. Koutnikova H, Oyasu R, Scarpelli DG. zo-2 gene alternative promoters in normal and neoplastic human pancreatic duct Familial hypercholanemia (FHC) cells. Int J Cancer. 1999 Oct 29;83(3):349-58 Note Fedele CG, Ciardi M, Delia S, Echevarria JM, Tenorio A. Mutation 143T-->C, predicted to cause a valine to Multiplex polymerase chain reaction for the simultaneous alanine substitution (V48A) in TJP2 produces the loss detection and typing of polyomavirus JC, BK and SV40 DNA in of the alpha-helical structure of PDZ1 domain of TJP2. clinical samples. J Virol Methods. 1999 Oct;82(2):137-44 This mutation reduces PDZ1 domain stability and Itoh M, Morita K, Tsukita S. Characterization of ZO-2 as a ligand binding in vitro. Thus binding of PDZ1 domain MAGUK family member associated with tight as well as adherens junctions with a binding affinity to occludin and alpha of TJP2 to peptides corresponding to the six C-terminal catenin. J Biol Chem. 1999 Feb 26;274(9):5981-6 amino acids from claudin 1, claudin 2, claudin 3, claudin 5 and claudin 7 is significantly reduced when Chlenski A, Ketels KV, Korovaitseva GI, Talamonti MS, Oyasu R, Scarpelli DG. Organization and expression of the human zo- V48A mutation is present. This mutation is associated 2 gene (tjp-2) in normal and neoplastic tissues. Biochim with Familial Hypercholanemia (FHC). Biophys Acta. 2000 Oct 2;1493(3):319-24 Disease Glaunsinger BA, Weiss RS, Lee SS, Javier R. Link of the Mutation 143T-->C, causing a valine to alanine unique oncogenic properties of adenovirus type 9 E4-ORF1 to substitution (V48A) in TJP2 is associated to Familial a select interaction with the candidate tumor suppressor protein ZO-2. EMBO J. 2001 Oct 15;20(20):5578-86 hypercholanemia (FCH). Huang HY, Li R, Sun Q, Wang J, Zhou P, Han H, Zhang WH. Familial hypercholanemia is characterized by elevated [LIM protein KyoT2 interacts with human tight junction protein serum bile acid concentrations, itching and fat ZO-2-i3]. Yi Chuan Xue Bao. 2002;29(11):953-8 malabsorption. In TJP2143C/143C individuals Islas S, Vega J, Ponce L, González-Mariscal L. Nuclear inheritance is oligogenic, with mutations in bile acid localization of the tight junction protein ZO-2 in epithelial cells. coenzyme A amino acid N-acyltransferase (BAAT), Exp Cell Res. 2002 Mar 10;274(1):138-48 required for clinical disease. Carlton VE, Harris BZ, Puffenberger EG, Batta AK, Knisely AS, Robinson DL, Strauss KA, Shneider BL, Lim WA, Salen G, To be noted Morton DH, Bull LN. Complex inheritance of familial hypercholanemia with associated mutations in TJP2 and Note BAAT. Nat Genet. 2003 May;34(1):91-6 TJP2 is proposed to be a tumor suppressor gene Sato N, Fukushima N, Maitra A, Matsubayashi H, Yeo CJ, because: 1) TJP2 protein and/or mRNA expression is Cameron JL, Hruban RH, Goggins M. Discovery of novel either lost or decreased in pancreatic, prostate, breast targets for aberrant methylation in pancreatic carcinoma using high-throughput microarrays. Cancer Res. 2003 Jul and lung adenocarcinomas, in testicular in situ 1;63(13):3735-42 carcinoma and in lung squamous carcinoma. 2) Has 40% homology to the tumor suppressor gene Dlg. 3) Traweger A, Fuchs R, Krizbai IA, Weiger TM, Bauer HC, Bauer H. The tight junction protein ZO-2 localizes to the nucleus and TJP2 is target of the major oncogenic determinant E4- interacts with the heterogeneous nuclear ribonucleoprotein ORF1 of human adenovirus type 9 (Ad9). E4-ORF1 scaffold attachment factor-B. J Biol Chem. 2003 Jan has a PDZ binding motif that sequesters TJP2 in the 24;278(4):2692-700 cytoplasm blocking its localization at the TJ. Betanzos A, Huerta M, Lopez-Bayghen E, Azuara E, Amerena Additionally the over-expression of TJP2 suppresses J, González-Mariscal L. The tight junction protein ZO-2 transformation by Ad9 E4-ORF-1, activated Ras V12 associates with Jun, Fos and C/EBP transcription factors in and the polyomavirus middle T protein. 4) TJP2 over- epithelial cells. Exp Cell Res. 2004 Jan 1;292(1):51-66 expression lowers the level of cyclin D1, blocks cell Humphray SJ, Oliver K, Hunt AR, Plumb RW, Loveland JE, cycle progression from G1 to S and inhibits cell Howe KL, Andrews TD, Searle S, Hunt SE, Scott CE, Jones MC, Ainscough R, Almeida JP, Ambrose KD, Ashwell RI, proliferation. Babbage AK, Babbage S, Bagguley CL, Bailey J, Banerjee R, Barker DJ, Barlow KF, Bates K, Beasley H, Beasley O, Bird References CP, Bray-Allen S, Brown AJ, Brown JY, Burford D, Burrill W, Burton J, Carder C, Carter NP, Chapman JC, Chen Y, Clarke Gumbiner B, Lowenkopf T, Apatira D. Identification of a 160- G, Clark SY, Clee CM, Clegg S, Collier RE, Corby N, Crosier kDa polypeptide that binds to the tight junction protein ZO-1. M, Cummings AT, Davies J, Dhami P, Dunn M, Dutta I, Dyer Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3460-4 LW, Earthrowl ME, Faulkner L, Fleming CJ, Frankish A,

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Frankland JA, French L, Fricker DG, Garner P, Garnett J, Fink C, Weigel R, Hembes T, Lauke-Wettwer H, Kliesch S, Ghori J, Gilbert JG, Glison C, Grafham DV, Gribble S, Griffiths Bergmann M, Brehm RH. Altered expression of ZO-1 and ZO-2 C, Griffiths-Jones S, Grocock R, Guy J, Hall RE, Hammond S, in Sertoli cells and loss of blood-testis barrier integrity in Harley JL, Harrison ES, Hart EA, Heath PD, Henderson CD, testicular carcinoma in situ. Neoplasia. 2006 Dec;8(12):1019- Hopkins BL, Howard PJ, Howden PJ, Huckle E, Johnson C, 27 Johnson D, Joy AA, Kay M, Keenan S, Kershaw JK, Kimberley AM, King A, Knights A, Laird GK, Langford C, Lawlor S, Umeda K, Ikenouchi J, Katahira-Tayama S, Furuse K, Sasaki Leongamornlert DA, Leversha M, Lloyd C, Lloyd DM, Lovell J, H, Nakayama M, Matsui T, Tsukita S, Furuse M, Tsukita S. Martin S, Mashreghi-Mohammadi M, Matthews L, McLaren S, ZO-1 and ZO-2 independently determine where claudins are McLay KE, McMurray A, Milne S, Nickerson T, Nisbett J, polymerized in tight-junction strand formation. Cell. 2006 Aug Nordsiek G, Pearce AV, Peck AI, Porter KM, Pandian R, Pelan 25;126(4):741-54 S, Phillimore B, Povey S, Ramsey Y, Rand V, Scharfe M, Huerta M, Muñoz R, Tapia R, Soto-Reyes E, Ramírez L, Sehra HK, Shownkeen R, Sims SK, Skuce CD, Smith M, Recillas-Targa F, González-Mariscal L, López-Bayghen E. Steward CA, Swarbreck D, Sycamore N, Tester J, Thorpe A, Cyclin D1 is transcriptionally down-regulated by ZO-2 via an E Tracey A, Tromans A, Thomas DW, Wall M, Wallis JM, West box and the transcription factor c-Myc. Mol Biol Cell. 2007 AP, Whitehead SL, Willey DL, Williams SA, Wilming L, Wray Dec;18(12):4826-36 PW, Young L, Ashurst JL, Coulson A, Blöcker H, Durbin R, Sulston JE, Hubbard T, Jackson MJ, Bentley DR, Beck S, Paschoud S, Bongiovanni M, Pache JC, Citi S. Claudin-1 and Rogers J, Dunham I. DNA sequence and analysis of human claudin-5 expression patterns differentiate lung squamous cell chromosome 9. Nature. 2004 May 27;429(6990):369-74 carcinomas from adenocarcinomas. Mod Pathol. 2007 Sep;20(9):947-54 Jaramillo BE, Ponce A, Moreno J, Betanzos A, Huerta M, Lopez-Bayghen E, Gonzalez-Mariscal L. Characterization of Sheth B, Nowak RL, Anderson R, Kwong WY, Papenbrock T, the tight junction protein ZO-2 localized at the nucleus of Fleming TP. Tight junction protein ZO-2 expression and epithelial cells. Exp Cell Res. 2004 Jul 1;297(1):247-58 relative function of ZO-1 and ZO-2 during mouse blastocyst formation. Exp Cell Res. 2008 Nov 1;314(18):3356-68 Kausalya PJ, Phua DC, Hunziker W. Association of ARVCF with zonula occludens (ZO)-1 and ZO-2: binding to PDZ- Xu J, Kausalya PJ, Phua DC, Ali SM, Hossain Z, Hunziker W. domain proteins and cell-cell adhesion regulate plasma Early embryonic lethality of mice lacking ZO-2, but Not ZO-3, membrane and nuclear localization of ARVCF. Mol Biol Cell. reveals critical and nonredundant roles for individual zonula 2004 Dec;15(12):5503-15 occludens proteins in mammalian development. Mol Cell Biol. 2008 Mar;28(5):1669-78 Martin TA, Watkins G, Mansel RE, Jiang WG. Loss of tight junction plaque molecules in breast cancer tissues is Tapia R, Huerta M, Islas S, Avila-Flores A, Lopez-Bayghen E, associated with a poor prognosis in patients with breast Weiske J, Huber O, González-Mariscal L. Zona occludens-2 cancer. Eur J Cancer. 2004 Dec;40(18):2717-25 inhibits cyclin D1 expression and cell proliferation and exhibits changes in localization along the cell cycle. Mol Biol Cell. 2009 Wang Y, Yu Q, Cho AH, Rondeau G, Welsh J, Adamson E, Feb;20(3):1102-17 Mercola D, McClelland M. Survey of differentially methylated promoters in prostate cancer cell lines. Neoplasia. 2005 This article should be referenced as such: Aug;7(8):748-60 Gonzalez-Mariscal L, Garay E, Quiros M, Tapia R. TJP2 (tight junction protein 2 (zona occludens 2)). Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4):423-428.

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Leukaemia Section Mini Review

+16 or trisomy 16 (solely) Jean-Loup Huret Genetics, Dept Medical Information, University of Poitiers, CHU Poitiers Hospital, F-86021 Poitiers, France (JLH)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Anomalies/tri16ID1537.html DOI: 10.4267/2042/44743 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Clinics and pathology Disease Solid tumours Disease Note Haematological malignancies. Trisomy 16 solely is a very rare anomaly in solid Note tumours; it has been detected in 2 cases of adeno- Trisomy 16 solely has been found in only 14 cases of carcinoma of the kidney (Gayrard et al., 2008; Kardas blood malignancies so far. et al., 2005), one adenocarcinoma of the prostate (Verdorfer et al., 2001), in one case of basal cell Phenotype/cell stem origin carcinoma of the skin (Casalone et al., 2000), in a case Acute lymphoblastic leukemia (B-ALL in 5 cases, ALL of squamous cell carcinoma of the vagina (Micci et al., not otherwise specified (NOS) in the 3 remaining 2003) and in a case of intraductal papilloma of the cases) (Ahmad et al., 2008; Arana-Trejo et al., 1993; breast (Lundin et al., 1998). Guillaume et al., 2001; Heerema et al., 1985; Silva et al., 2002; Testa et al., 1985; Tsang et al., 2001; Yamada Cytogenetics and Furusawa, 1976), acute myeloid leukemia (one M2-AML, one AML-NOS) (Hda et al., 1996; Li et al., Cytogenetics molecular 1983), myelodysplastic syndromes (one case of To be noted is that a cryptic ETV6/RUNX1 was found refractory anemia (RA) and 2 cases of refractory in one case of common ALL (Tsang et al., 2001). anemia with excess of blasts (RAEB)) (Horiike et al., 1988; Guillaume et al., 2001), myeloproliferative Genes involved and proteins syndromes (idiopathic myelofibrosis, one case) (Whang-Peng et al., 1978), chronic lymphocytic Note leukemia (2 cases, Ohtaki et al., 1986; Sadamori et al., Genes involved in the process giving rise to the trisomy 1984) and Hodgkin disease (2 cases, Banks et al., 1991; 16, as well as the consequences of trisomy 16 in terms Kristoffersson et al., 1987). of carcinogenesis are unknown. Epidemiology References The ALL cases were 4M/3F, aged 2, 5, 18, 24, 32 and 35 years. The myeloid cases were 2M/3F, aged 1, 26, Yamada K, Furusawa S. Preferential involvement of chromosomes no. 8 and no. 21 in acute leukemia and 33, 49 and 61 years. preleukemia. Blood. 1976 Apr;47(4):679-86 Prognosis Whang-Peng J, Lee E, Knutsen T, Chang P, Nienhuis A.. Survival in myeloid cases was the following: patients Cytogenetic studies in patients with myelofibrosis and myeloid deceased at 11, 33, 52 and 60 months (the latter in the metaplasia. Leukemia Res. 1978;2:41-56. idiopathic myelofibrosis case), one Li YS, Khalid G, Hayhoe FG. Correlation between chromosomal pattern, cytological subtypes, response to patient was still alive at 22 months. Data is even therapy, and survival in acute myeloid leukaemia. Scand J scarcer in the ALL cases (10, 24+, 58+). Haematol. 1983 Mar;30(3):265-77

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Sadamori N, Han T, Minowada J, Sandberg AA. cell carcinomas: different findings in direct preparations and Chromosomes and causation of human cancer and leukemia. short-term cell cultures. Cancer Genet Cytogenet. 2000 Apr LII. Chromosome findings in treated patients with B-cell chronic 15;118(2):136-43 lymphocytic leukemia. Cancer Genet Cytogenet. 1984 Feb;11(2):161-8 Guillaume B, Ameye G, Dierlamm J, Verhoef G, Duhem C, Ferrant A, Hagemeijer A, Verellen-Dumoulin C, Michaux L. Heerema NA, Palmer CG, Baehner RL. Karyotypic and clinical Trisomy 16 as the sole anomaly in hematological findings in a consecutive series of children with acute malignancies. Three new cases and a short review. Cancer lymphocytic leukemia. Cancer Genet Cytogenet. 1985 Genet Cytogenet. 2001 Jul 15;128(2):168-71 Jun;17(2):165-79 Tsang KS, Li CK, Chik KW, Shing MM, Tsoi WC, Ng MH, Lau Testa JR, Misawa S, Oguma N, Van Sloten K, Wiernik PH. TT, Leung Y, Yuen PM. TEL/AML1 rearrangement and the Chromosomal alterations in acute leukemia patients studied prognostic significance in childhood acute lymphoblastic with improved culture methods. Cancer Res. 1985 leukemia in Hong Kong. Am J Hematol. 2001 Oct;68(2):91-8 Jan;45(1):430-4 Verdorfer I, Hobisch A, Culig Z, Hittmair A, Bartsch G, Erdel M, Ohtaki K, Han T, Sandberg AA. Sequential chromosome Duba HC, Utermann G. Combined study of prostatic carcinoma abnormalities in B cell chronic lymphocytic leukemia: a study of by classical cytogenetic analysis and comparative genomic 13 cases. Cancer Genet Cytogenet. 1986 Feb 1;20(1-2):73-87 hybridization. Int J Oncol. 2001 Dec;19(6):1263-70 Kristoffersson U, Heim S, Mandahl N, Olsson H, Akerman M, Silva ML, Ornellas de Souza MH, Ribeiro RC, Land MG, Mitelman F.. Cytogenetic studies in Hodgkin's disease. APMIS. Boulhosa de Azevedo AM, Vasconcelos F, Otero L, 1987;95:289-95. Vasconcelos Z, Bouzas LF, Abdelhay E. Cytogenetic analysis of 100 consecutive newly diagnosed cases of acute Horiike S, Taniwaki M, Misawa S, Abe T. Chromosome lymphoblastic leukemia in Rio de Janeiro. Cancer Genet abnormalities and karyotypic evolution in 83 patients with Cytogenet. 2002 Sep;137(2):85-90 myelodysplastic syndrome and predictive value for prognosis. Cancer. 1988 Sep 15;62(6):1129-38 Micci F, Teixeira MR, Scheistrøen M, Abeler VM, Heim S. Cytogenetic characterization of tumors of the vulva and vagina. Banks RE, Gledhill S, Ross FM, Krajewski A, Dewar AE, Weir- Genes Chromosomes Cancer. 2003 Oct;38(2):137-48 Thompson EM. Karyotypic abnormalities and immunoglobulin gene rearrangements in Hodgkin's disease. Cancer Genet Kardas I, Mrózek K, Babinska M, Krajka K, Hadaczek P, Cytogenet. 1991 Jan;51(1):103-11 Lubinski J, Roszkiewicz A, Kuziemska E, Limon J. Cytogenetic and molecular findings in 75 clear cell renal cell carcinomas. Arana-Trejo RM, Cervantes-Peredo A, Rozen E, Kassack JJ, Oncol Rep. 2005 May;13(5):949-56 Gutierrez M, Kofman-Alfaro S.. Estudio citogenetico en 22 adultos y tres ninos con leucemia linfoblastica aguda. Rev Inv Ahmad F, Dalvi R, Chavan D, Das BR, Mandava S. Clin. 1993;45:43-8. Cytogenetic profile of acute lymphocytic leukemia patients: report of a novel translocation t(4;13) (q21 x 3; q35) from an Hda N, Chadli B, Bousfiha A, Trachli A, Harif M, Benslimane A. Indian population. Hematology. 2008 Feb;13(1):28-33 Cytogenetic survey of 53 Moroccan patients with acute myeloblastic leukemia. Cancer Genet Cytogenet. 1996 Gayrard N, Cacheux V, Iborra F, Mourad G, Argilés A. Feb;86(2):124-8 Cytogenetic studies of 24 renal epithelial tumors with von Hippel-Lindau and fragile histidine triad protein expression Lundin CP, Mertens F, Ingvar C, Idvall I, Pandis N. Trisomy 16 correlation. Arch Pathol Lab Med. 2008 Jun;132(6):965-73 as the primary chromosome aberration in a papilloma of the breast. Cancer Genet Cytogenet. 1998 Oct 1;106(1):90-1 This article should be referenced as such: Casalone R, Mazzola D, Righi R, Granata P, Minelli E, Huret JL. +16 or trisomy 16 (solely). Atlas Genet Cytogenet Salvadore M, Lombardo M, Bertani E. Cytogenetic and Oncol Haematol. 2010; 14(4):429-430. interphase FISH analyses of 73 basal cell and three squamous

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Leukaemia Section Short Communication t(3;3)(q27;q29) Jean-Loup Huret Genetics, Dept Medical Information, University of Poitiers, CHU Poitiers Hospital, F-86021 Poitiers, France (JLH)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Anomalies/t0303q27q29ID2087.html DOI: 10.4267/2042/44744 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

repressing complex, another transcription repression Clinics and pathology domain (191-386), PEST sequences (300-417) with a Disease KKYK motif (375-379), and six zinc finger at the C- term (518-541, 546-568, 574-596, 602-624, 630-652, Non Hodgkin lymphoma (NHL) 658-681), responsible for sequence specific DNA Epidemiology binding. Transcription repressor; recognizes the Four cases to date: a follicular mixed cell lympho-ma consensus sequence: TTCCT(A/C)GAA (Albagli- in a 78-year-old female patient, a grade 3B follicular Curiel, 2003). lymphoma in a female patient, a gastric lymphoma, and TFRC a diffuse large B-cell lymphoma (DLCL) (Yunis et al., Location 1984; Bosga-Bouwer et al., 2003; Chen et al., 2006; 3q29 Yoshida et al., 1999). Protein Prognosis 760 amino acids; composed of an NH2-term No data. cytoplasmic domain (amino acids 1-67), a trans- membrane domain (aa 68-88), and a C term Cytogenetics extracellular domain (aa 89-706) according to Swiss- Prot. Cell surface membrane glycoprotein; involved in Additional anomalies iron homeostasis by regulating cellular iron uptake There was a complex karyotype, including a (Dorak, 2008). t(14;18)(q32;q21), +15, and other anomalies in one case (Yunis et al., 1984), and also a complex karyotype Result of the chromosomal in an other case (Bosga-Bouwer et al., 2003); data is missing in the two other cases. anomaly Genes involved and proteins Hybrid gene Description BCL6 5' TFRC - 3' BCL6, with a breakpoint in BCL6 Location between exon 1 and 2 (Yoshida et al., 1999); the 3q27 breakpoint on TFRC was found 288 bp 3' downstream of the breakpoint in the other case studied with Protein molecular techniques (Chen et al., 2006). 706 amino acids; composed of a NH2-term BTB/POZ domain (amino acids 1-130 (32-99 according to Swiss- References Prot)) which mediates homodi-merization and protein- protein interactions with other corepressors (including Yunis JJ, Oken MM, Theologides A, Howe RB, Kaplan ME. Recurrent chromosomal defects are found in most patients HDAC1 and NCOR2/ SMRT) to constitute a large with non-Hodgkin's-lymphoma. Cancer Genet Cytogenet. 1984 Sep;13(1):17-28

Atlas Genet Cytogenet Oncol Haematol. 2010; 14(4) 431 t(3;3)(q27;q29) Huret JL

Yoshida S, Kaneita Y, Aoki Y, Seto M, Mori S, Moriyama M. Chen YW, Hu XT, Liang AC, Au WY, So CC, Wong ML, Shen Identification of heterologous translocation partner genes fused L, Tao Q, Chu KM, Kwong YL, Liang RH, Srivastava G. High to the BCL6 gene in diffuse large B-cell lymphomas: 5'-RACE BCL6 expression predicts better prognosis, independent of and LA - PCR analyses of biopsy samples. Oncogene. 1999 BCL6 translocation status, translocation partner, or BCL6- Dec 23;18(56):7994-9 deregulating mutations, in gastric lymphoma. Blood. 2006 Oct 1;108(7):2373-83 Albagli-Curiel O. Ambivalent role of BCL6 in cell survival and transformation. Oncogene. 2003 Jan 30;22(4):507-16 Dorak MT.. TFRC (transferrin receptor (p90, CD71)). Atlas Genet Cytogenet Oncol Haematol. April 2008. URL: Bosga-Bouwer AG, van Imhoff GW, Boonstra R, van der Veen http://AtlasGeneticsOncology.org/Genes/TFRCID259ch3q29.ht A, Haralambieva E, van den Berg A, de Jong B, Krause V, ml. Palmer MC, Coupland R, Kluin PM, van den Berg E, Poppema S. Follicular lymphoma grade 3B includes 3 cytogenetically This article should be referenced as such: defined subgroups with primary t(14;18), 3q27, or other translocations: t(14;18) and 3q27 are mutually exclusive. Huret JL. t(3;3)(q27;q29). Atlas Genet Cytogenet Oncol Blood. 2003 Feb 1;101(3):1149-54 Haematol. 2010; 14(4):431-432.

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Leukaemia Section Short Communication t(8;9)(q24;q13) Jean-Loup Huret Genetics, Dept Medical Information, University of Poitiers, CHU Poitiers Hospital, F-86021 Poitiers, France (JLH)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Anomalies/t0809q24q13ID1550.html DOI: 10.4267/2042/44745 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Clinics and pathology including +7, +17, +19, i(6p) and other anomalies. Disease Genes involved and proteins Non Hodgkin lymphoma and acute lymphoblastic Note lymphoma. In the follicular lymphoma case with t(1;14)(q21;q32), Phenotype/cell stem origin there was deregulation of FCGR2B expression due to FCGR2B/IgH juxta-position. B-cell malignancies. Epidemiology References 3 cases to date: a stage IIIA mantle cell lymphoma in a Au WY, Horsman DE, Viswanatha DS, Connors JM, Klasa RJ, male aged 58 years (Au et al., 2000; Au et al., 2002), a Gascoyne RD. 8q24 translocations in blastic transformation of follicular lymphoma in a female patient aged 44 years mantle cell lymphoma. Haematologica. 2000 Nov;85(11):1225- (Chen et al., 2001), and a pre-B acute lymphoblastic 7 leukemia (ALL) in a female patient aged 60 years Chen W, Palanisamy N, Schmidt H, Teruya-Feldstein J, (Pedersen et al., 2001). Jhanwar SC, Zelenetz AD, Houldsworth J, Chaganti RS. Deregulation of FCGR2B expression by 1q21 rearrangements Evolution in follicular lymphomas. Oncogene. 2001 Nov 15;20(52):7686- 93 Clinical data is available in the mantle cell lymphoma case, who reached complete remission, relapsed 2 years Pedersen RK, Kerndrup GB, Sørensen AG, Mourits-Andersen T, Gram-Hansen P, Pulczynski S, Schmidt KG. Cytogenetic later, and died. aberrations in adult acute lymphocytic leukemia: optimal technique may influence the results. Cancer Genet Cytogenet. Cytogenetics 2001 Jul 1;128(1):7-10 Au WY, Gascoyne RD, Viswanatha DS, Connors JM, Klasa Additional anomalies RJ, Horsman DE. Cytogenetic analysis in mantle cell The mantle cell lymphoma case exhibited a complex lymphoma: a review of 214 cases. Leuk Lymphoma. 2002 karyotype with a t(11;14)(q13;q32), a t(9;14)(q13;q32) Apr;43(4):783-91 and other anomalies, the follicular lymphoma a This article should be referenced as such: t(1;14)(q21;q32), and the ALL case a complex Huret JL. t(8;9)(q24;q13). Atlas Genet Cytogenet Oncol karyotype, including +7, +12, add(14)(q32), and other Haematol. 2010; 14(4):433. anomalies. The follicular lymphoma case progressed towards a diffuse large cell lymphoma with a complex karyotype,

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Solid Tumour Section Short Communication t(2;22)(q31;q12) in a small round cell tumour Jean-Loup Huret Genetics, Dept Medical Information, University of Poitiers, CHU Poitiers Hospital, F-86021 Poitiers, France (JLH)

Published in Atlas Database: May 2009 Online updated version: http://AtlasGeneticsOncology.org/Tumors/t0222q31q12SmallRoundID6278.html DOI: 10.4267/2042/44746 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology

(TAD) containing multiple degenerate hexapeptide Clinics and pathology repeats, 3 arginine/glycine rich domains (RGG Disease regions), a RNA recognition motif, and a RanBP2 type Zinc finger. Role in transcriptional regulation for A case of undifferentiated small round cell tumour of specific genes and in mRNA splicing. the frontal region, with multiple metastases at diagnosis (in long bones, lungs, kidney), was described in a 16- Result of the chromosomal year-old male patient. The patient died 20 months after diagnosis (Wang et al., 2007). anomaly Cytogenetics Hybrid Gene Description Cytogenetics Morphological 5' EWSR1 - 3' SP3. Fusion of EWSR1 exon 7 to exon 6 The t(2;22)(q31;q12) was the sole anomaly. of SP3. Genes involved and proteins Fusion Protein Description SP3 The N terminal transactivation domain of EWSR1 was Location fused to the zinc fingers (DNA binding domain) of 2q31 SP3. Protein References From N-term to C-term: a transactivation domain, a repressor domain, and 3 C2H2 zinc finger domains. Wang L, Bhargava R, Zheng T, Wexler L, Collins MH, Roulston Transcription factor. D, Ladanyi M. Undifferentiated small round cell sarcomas with rare EWS gene fusions: identification of a novel EWS-SP3 EWSR1 fusion and of additional cases with the EWS-ETV1 and EWS- FEV fusions. J Mol Diagn. 2007 Sep;9(4):498-509 Location 22q12 This article should be referenced as such: Protein Huret JL. t(2;22)(q31;q12) in a small round cell tumour. Atlas From N-term to C-term: a transactivation domain Genet Cytogenet Oncol Haematol. 2010; 14(4):434.

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