Number 9 September 2014

Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Volume 18 - Number 9 September 2014

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 in Oncology and Haematology

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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, and also more traditional review articles (“deep insights”) on the above subjects and on surrounding topics. It also present 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, Marie-Christine Jacquemot-Perbal, Vanessa Le Berre, Anne Malo, Carol Moreau, Catherine Morel-Pair, Laurent Rassinoux, 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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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 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 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 Adriana Zamecnikova (Kuwait) Leukaemia Section

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Volume 18, Number 9, September 2014

Table of contents

Gene Section

AFAP1L2 (actin filament associated protein 1-like 2) 628 Xiaohui Bai, Serisha Moodley, Hae-Ra Cho, Mingyao Liu ENOX2 (ecto-NOX disulfide-thiol exchanger 2) 632 Xiaoyu Tang, Dorothy M Morré, D James Morré FABP7 (fatty acid binding protein 7, brain) 638 Roseline Godbout, Ho-Yin Poon, Rong-Zong Liu GSTA1 (glutathione S-transferase alpha 1) 645 Ana Savic-Radojevic, Tanja Radic MOAP1 (Modulator Of Apoptosis 1) 650 Gamze Ayaz, Mesut Muyan PHLDA1 (pleckstrin homology-like domain, family A, member 1) 652 Maria Aparecida Nagai ADAMTS15 (ADAM Metallopeptidase With Thrombospondin Type 1 Motif, 15) 655 Santiago Cal, Alvaro J Obaya ADRB2 (adrenoceptor beta 2, surface) 659 Denise Tostes Oliveira, Diego Mauricio Bravo-Calderón IRF4 (interferon regulatory factor 4) 663 Vipul Shukla, Runqing Lu PLCG1 (Phospholipase C, Gamma 1) 668 Rebeca Manso SLC1A5 (solute carrier family 1 (neutral amino acid transporter), member 5) 673 Cesare Indiveri, Lorena Pochini, Michele Galluccio, Mariafrancesca Scalise USB1 (U6 snRNA biogenesis 1) 678 Elisa Adele Colombo

Leukaemia Section t(9;15)(p13;q24) PAX5/PML 682 Jean-Loup Huret t(1;9)(p13;p12) PAX5/HIPK1 685 Jean-Loup Huret

Solid Tumour Section

Lung: t(6;12)(q22;q14.1) LRIG3/ROS1 in lung adenocarcinoma 688 Kana Sakamoto, Yuki Togashi, Kengo Takeuchi

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Deep Insight Section

The tumour suppressor function of the scaffolding protein spinophilin 691 Denis Sarrouilhe, Véronique Ladeveze

Case Report Section

Translocation t(5;6)(q33-34;q23) in an acute myelomonocytic leukemia patient 701 Adriana Zamecnikova, Soad Al Bahar, Ramesh Pandita

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Gene Section Review

AFAP1L2 (actin filament associated protein 1- like 2) Xiaohui Bai, Serisha Moodley, Hae-Ra Cho, Mingyao Liu Latner Thoracic Surgery Research Laboratoires, University Health Network, Toronto General Research Institute, University of Toronto, Toronto, Ontario, Canada (XB, SM, HRC, ML)

Published in Atlas Database: January 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/AFAP1L2ID52197ch10q25.html DOI: 10.4267/2042/54026 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract Identity Review on AFAP1L2, with data on DNA/RNA, on Other names: KIAA1914, XB130 the protein encoded and where the gene is HGNC (Hugo): AFAP1L2 implicated. Location: 10q25.3

Figure 1. XB130 chromosomal location and neighbour genes. A. xb130 gene is located on chromosome 10, at 10q25.3 by fluorescence in situ hybridization (FISH). B. Diagram of xb130 neighbour genes between 115939029 and 116450393.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 628 AFAP1L2 (actin filament associated protein 1-like 2) Bai X, et al.

Figure 2. XB130 functional domains and motifs. Human XB130 has 818 amino acids. It contains the following motif/domains: proline-rich region: residues 98-107; tyrosine phosphorylation motif: residues 54-57, 124-127; 148-151; 457-460; PH domain: residues 175-272; 353-445; coiled-coil region: residues 652-749.

approximately 130 kDa by western blotting (Xu et DNA/RNA al., 2007). As an adaptor protein, XB130 has no Note enzymatic domains or activity. Sequence structure Human XB130 was discovered in Dr. Mingyao analysis has revealed 23 putative tyrosine Liu's laboratory (University of Toronto) in the phosphorylation sites and 27 putative process of cloning human actin filament associated phosphorylation sites for serine/threonine kinases protein (afap) gene. Using chicken AFAP protein (Xu et al., 2007). The N-terminal of XB130 sequence to search human cDNA library in contains a proline rich, SH3 domain binding motif, GenBank, XB130 was found as an EST clone three tyrosine containing SH2 domain binding sites (GenBank accession number 1154093) with 34% (Xu et al., 2007), of which a YXXM motif is for sequence similarity to chicken AFAP protein. The PI3 kinase subunit p85 binding (Lodyga et al., clone contains partial coding sequence and 3' UTR. 2009). In the middle region, there are two pleckstrin The upstream sequence was obtained using 5' rapid homology domains and another tyrosine binding amplification of cDNA ends (RACE) from human motif (Xu et al., 2007). The C-terminal contains a lung alveolar epithelial cell mRNA. Western blot coiled-coil region, which may be important for shows the protein molecular weight is 130 kD (Xu molecular trafficking or dimerization (Xu et al., et al., 2007). 2007). In 2003, the XB130 knockout mice were Expression established through the collaboration of Drs. Mingyao Liu and Tak W. Mak at the University of In normal human tissue, the 4 kb mRNA transcript Toronto. of XB130 is expressed highly in spleen and thyroid with lower expression in kidney, brain, lung and Description pancreas (Xu et al., 2007). Human xb130 genes contains 19 exons, which are Newer RNA sequencing by Illumina body map covering the whole coding sequence. using RNA obtained from 16 normal human tissues shows high expression of XB130 in thyroid with Transcription lower expression in lymph nodes, brain, colon, The transcript size of xb130 is 3751 bp. There may adipocytes, kidney, lung, adrenal glands, breast, be 7 splicing variants based on Ensembl data ovary, prostate and testis followed by whole blood, (www.ensembl.org). XB130 mRNA is highly heart, skeletal muscles and liver expressed in the thyroid, parathyroid and spleen; (www.genecards.org). moderately expressed in brain, pancreas, lung and XB130 protein is detected in normal tissues of kidney. thyroid, parathyroid, brain, kidney, skin and GI- tracks (www.proteinatlas.org). Protein XB130 protein expresses in human thyroid, Description colorectal, gastric and hepatocellular carcinomas (Shi et al., 2012; Shiozaki et al., 2013; Shiozaki et XB130 is a novel adaptor protein, member of the al., 2011; Zuo et al., 2012). Expression of XB130 actin filament associated protein (AFAP) family has also been observed in a variety of cancer cell (Snyder et al., 2011). Accordingly, it is also known lines, including thyroid, lung, esophageal, as AFAP1L2. The full length protein consists of pancreatic and colon cancers (Shi et al., 2012; 818 amino acids with a molecular weight of Shiozaki et al., 2013; Zuo et al., 2012).

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 629 AFAP1L2 (actin filament associated protein 1-like 2) Bai X, et al.

Localisation lung, large intestine, ovary, skin, prostate, endometrium. XB130 is distributed mainly in the cytoplasm and Among these samples, 70% of identified cases are perinuclear region of lung epithelial BEAS-2B cells XB130 substitution missense mutation. and several other cell types (Xu et al., 2007). Unlike AFAP, XB130 does not associate or co- localize with actin filament stress fibler (Lodyga et Implicated in al., 2010). Various cancers Stimulation of cells with EGF, PMA, or overexpression of constitutive Rac results in a Note translocation of XB130 to the cell periphery and XB130 plays important roles in tumor progression leading edge of migrating cells (Lodyga et al., by promoting cell proliferation, survival, motility 2010). and invasion in various cancer cells. Recently, XB130 has been identified in thyroid Function carcinoma (Shiozaki et al., 2011), esophageal XB130 is an adaptor protein that acts as a key squamous cell carcinoma (Shiozaki et al., 2013), mediator to drive signal transduction pathways. and gastric cancer (Shi et al., 2012). XB130 has been shown to bind to tyrosine kinase c- Src to enhance kinase activity and subsequently Colorectal cancer regulates Src-mediated AP-1/SRE transcription Note activation (Xu et al., 2007). Tyrosine phosphorylated XB130 in colorectal XB130 is also highly involved in the PI3K/Akt cancer. pathway and effects cell proliferation, cell cycle Prognosis progression and cell survival through binding to Using mass spectrometry, Emaduddin et al. p85 alpha subunit of PI3K (Lodyga et al., 2009). reported several proteins as tyrosine phosphorylated XB130 may also play a role in the innate immune form are maintained at high level in colorectal response, where knockdown of XB130 was shown cancer cells isolated from patients. to decrease IL-6 and IL-8 cytokine levels in human XB130 is identified as one of these proteins. lung epithelial cells (Xu et al., 2007). XB130 is also Therefore, tyrosine phosphorylated XB130 has a involved in cell migration via association with Rac- potential to be a biomarker of colorectal cancer GTPase and plays a significant role in lateral cell (Emaduddin et al., 2008). migration and cell invasion in both normal and cancer cell lines (Lodyga et al., 2010). Gastric cancer (GC) Yamanaka et al. reported phosphorylated XB130 Note affects cAMP-dependent DNA synthesis in rat XB130 expression level associates with the thyroid cells (Yamanaka et al., 2012). XB130 is prognosis of gastric cancer. aberrantly expressed in human cancers and has been Prognosis shown to control tumour growth in vivo (Shiozaki Based on the anlysis GC tissue samples from 411 et al., 2011). patients with various stages, lower expression of XB130 regulates thyroid cancer cell proliferation XB130 mRNA as well as protein is significantly by controlling microRNA miR-33a, 149a and 193a correlated with reduced patient survival time and expression to alter oncogenes Myc, FosL1, and shorter disease-free period (Shi et al., 2012). SCL7A5 protein levels (Takeshita et al., 2013). Homology Thyroid cancer XB130 shares similar sequence and domain Note structure cellular as AFAP and AFAP1L1 (Snyder XB130 as a tumor promoting gene, enhancing et al., 2011). thyroid cancer cell growth. Oncogenesis Mutations Knockdown XB130 using siRNA in thyroid cancer cell (WRO) is accompanied with an inhibition of Somatic G1-S phase cell cycle progression and enhanced Somatic mutations of XB130 are reported in a apoptosis. variety of cancer tissues. The volume of tumors generated in nude mice after Based on the data of Sanger Institute database injecting these cells are smaller than those formed (www.sanger.ac.uk), XB130 mutation sites have from cells with a normal XB130 expression been identified in multiple tumor tissues, including (Shiozaki et al., 2011).

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 630 AFAP1L2 (actin filament associated protein 1-like 2) Bai X, et al.

Esophageal squamous cell is a Rac-controlled component of lamellipodia that regulates cell motility and invasion. J Cell Sci. 2010 Dec carcinoma (ESCC) 1;123(Pt 23):4156-69 Note Shiozaki A, Liu M. Roles of XB130, a novel adaptor XB130 protein is identified in ESCC primary cell protein, in cancer. J Clin Bioinforma. 2011 Mar 17;1(1):10 lines and tumor samples. Shiozaki A, Lodyga M, Bai XH, Nadesalingam J, Oyaizu T, Oncogenesis Winer D, Asa SL, Keshavjee S, Liu M. XB130, a novel adaptor protein, promotes thyroid tumor growth. Am J XB130 protein is highly expression in ESCC Pathol. 2011 Jan;178(1):391-401 primary cells. XB130 protein is examined by Snyder BN, Cho Y, Qian Y, Coad JE, Flynn DC, Cunnick immunohistochemistry staining from ESCC tissues JM. AFAP1L1 is a novel adaptor protein of the AFAP collected from 52 patients. Positive XB130 staining family that interacts with cortactin and localizes to is observed in 71% of ESCC samples, which invadosomes. Eur J Cell Biol. 2011 May;90(5):376-89 indicates the association of XB130 protein and Shi M, Huang W, Lin L, Zheng D, Zuo Q, Wang L, Wang ESCC (Shiozaki et al., 2013). N, Wu Y, Liao Y, Liao W. Silencing of XB130 is associated with both the prognosis and chemosensitivity of gastric Soft tissue tumor cancer. PLoS One. 2012;7(8):e41660 Note Shiozaki A, Shen-Tu G, Bai X, Iitaka D, De Falco V, Decreased XB130 expression leads to a local Santoro M, Keshavjee S, Liu M. XB130 mediates cancer aggressiveness of soft tissue tumor. cell proliferation and survival through multiple signaling events downstream of Akt. PLoS One. 2012;7(8):e43646 Oncogenesis Yamanaka D, Akama T, Fukushima T, Nedachi T, Analysis of gene expression profile of 102 tumor Kawasaki C, Chida K, Minami S, Suzuki K, Hakuno F, samples with varying stages of soft tissue tumor Takahashi S. Phosphatidylinositol 3-kinase-binding shows a decreased XB130 expression in malignant protein, PI3KAP/XB130, is required for cAMP-induced mesenchymal tumors (Cunha et al., 2010). amplification of IGF mitogenic activity in FRTL-5 thyroid cells. Mol Endocrinol. 2012 Jun;26(6):1043-55 References Zuo Q, Huang H, Shi M, Zhang F, Sun J, Bin J, Liao Y, Liao W. Multivariate analysis of several molecular markers Xu J, Bai XH, Lodyga M, Han B, Xiao H, Keshavjee S, Hu and clinicopathological features in postoperative prognosis J, Zhang H, Yang BB, Liu M. XB130, a novel adaptor of hepatocellular carcinoma. Anat Rec (Hoboken). 2012 protein for signal transduction. J Biol Chem. 2007 Jun Mar;295(3):423-31 1;282(22):16401-12 Shiozaki A, Kosuga T, Ichikawa D, Komatsu S, Fujiwara H, Emaduddin M, Edelmann MJ, Kessler BM, Feller SM. Odin Okamoto K, Iitaka D, Nakashima S, Shimizu H, Ishimoto T, (ANKS1A) is a Src family kinase target in colorectal cancer Kitagawa M, Nakou Y, Kishimoto M, Liu M, Otsuji E. cells. Cell Commun Signal. 2008 Oct 9;6:7 XB130 as an independent prognostic factor in human esophageal squamous cell carcinoma. Ann Surg Oncol. Lodyga M, De Falco V, Bai XH, Kapus A, Melillo RM, 2013 Sep;20(9):3140-50 Santoro M, Liu M. XB130, a tissue-specific adaptor protein that couples the RET/PTC oncogenic kinase to PI 3-kinase Takeshita H, Shiozaki A, Bai XH, Iitaka D, Kim H, Yang pathway. Oncogene. 2009 Feb 19;28(7):937-49 BB, Keshavjee S, Liu M. XB130, a new adaptor protein, regulates expression of tumor suppressive microRNAs in Cunha IW, Carvalho KC, Martins WK, Marques SM, Muto cancer cells. PLoS One. 2013;8(3):e59057 NH, Falzoni R, Rocha RM, Aguiar S, Simoes AC, Fahham L, Neves EJ, Soares FA, Reis LF. Identification of genes This article should be referenced as such: associated with local aggressiveness and metastatic behavior in soft tissue tumors. Transl Oncol. 2010 Bai X, Moodley S, Cho HR, Liu M. AFAP1L2 (actin filament Feb;3(1):23-32 associated protein 1-like 2). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):628-631. Lodyga M, Bai XH, Kapus A, Liu M. Adaptor protein XB130

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 631 Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Gene Section Review

ENOX2 (ecto-NOX disulfide-thiol exchanger 2) Xiaoyu Tang, Dorothy M Morré, D James Morré MorNuCo, Inc., 1201 Cumberland Avenue, Ste. B, Purdue Research Park,.West Lafayette, IN 47906 USA (XT, DMM, DJM)

Published in Atlas Database: January 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/ENOX2ID40134chXq26.html DOI: 10.4267/2042/54027 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

according to NCBI Refseq Gene Database (gene Abstract ID: 10495, RefSeq ID: NG_012562.1), and is Review on ENOX2, with data on DNA/RNA, on comprised of 279939 bp. the protein encoded and where the gene is ENOX2 is composed of 13 protein-coding exons implicated. between 71 bp and 2066 bp in length and 14 introns which vary greatly in length (1781 bp to 117994 Identity bp). It has a 501 bp 5' untranslated region and a long 3' Other names: APK1, COVA1, tNOX UTR (approximately 1935 bp). HGNC (Hugo): ENOX2 Transcription Location: Xq26.1 According to NCBI the human ENOX2 gene Note encodes a 4036 bp mRNA transcript, the coding Also termed APK1 antigen, or cytosolic ovarian sequence (CDS) located from base pairs 356 to carcinoma antigen 1, or tumor-associated 2101 (NM_001281736.1). hydroquinone oxidase (tNOX). The CDS from the Ensembl genome browser ECTO-NOX2 = Ecto-Nicotinamide Dinucleotide database (ENST00000370927, transcript length Oxidase Disulfide Thiol Exchange 2. 3788 bp) and NCBI (NM_001281736.1) are identical. DNA/RNA Transcripts NM_001281736.1 and ENST 00000370927 are also included in the human CCDS Description set (CCDS14626) and encode a 610 aa long protein. The human ENOX2 gene is located on the reverse Pseudogene strand of chromosome X (bases 4918 to 284856); None known.

Figure 1. ENOX2 mRNA.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 632 ENOX2 (ecto-NOX disulfide-thiol exchanger 2) Tang X, et al.

Figure 2. Deduced amino acid sequence and functional motifs of the bacterially expressed 46 kDa enzymatically active C- terminus of ENOX2.

for protein disulfide-thiol interchange (Morré and Protein Morré, 2013). Description The signature ENOX2 motif is that of the potential drug/antibody binding site E394EMTE. Antisera ENOX2 transcription variants all appear to be directed to this portion of the protein act as variations that include an exon 4 minus splicing competitive inhibitors to drug binding. The event that allows for down-stream initiation and sequence provides a putative quinone or expression of the ENOX2 protein at the cell surface sulfonylurea-binding site with four of the five of malignant cells (Tang et al., 2007a; Tang et al., amino acids in at least one other putative quinone 2007b). Without the exon 4 deletion, mRNA site in the same relative positions. derived from the gene does not appear to be The correctness of the various assignments has, for translated into protein. Thus, the exon 4 deletion is the most part, been confirmed by site-directed the basis for the cancer specificity of the ENOX2 mutagenesis (Chueh et al., 2002). transcription variants. An hnRNP splicing factor While amino acid replacements that block oxidation directs formation of the Exon 4 minus variants of of reduced pyridine nucleotide by ENOX2 also ENOX2 (Tang et al., 2011). The fully processed 34 eliminated protein disulfide-thiol interchange and kDa generic ENOX2 protein found on the cell vice versa (Chueh et al., 2002), the two activities surface of HeLa cells and in sera of about 23% of appear to occur independently. early cancer patients retains full-functional activity. One can be measured in the absence of the other. The deduced amino acid sequence of a bacterially The ENOX2 proteins have properties of prions and expressed 46 kDa functional C-terminus of ENOX2 are protease resistant (Kelker et al., 2001) and N- exhibits the same characteristics of alternation of terminal sequencing. the two activities and drug response as the cell Concentrated solutions aggregate and form surface and generic serums forms. Identified amyloid-like filaments. functional motifs include a quinone binding site, an adenine nucleotide binding site, a CXXXXC Expression cysteine motif as a potential disulfide-thiol Widely expressed in malignant cells but only as interchange site and two copper binding sites, one exon 4 minus splicing variants (Tang et al., 2007b). of which is conserved with superoxide dismutase. ENOX2 proteins lack flavin and only one of the Localisation two C-X-X-X-X-C motifs characteristic of External cell surface (Morré, 1995). flavoproteins are present in ENOX2. Yet the protein effectively carries out protein disulfide Function interchange. The motif C569-X-X-X-X-X-C575, ENOX2 is a member of a family of cell surface alone or together with a downstream histidine metalocatalysts with binuclear copper centers that (H582) provides an additional potential active site oscillate.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 633 ENOX2 (ecto-NOX disulfide-thiol exchanger 2) Tang X, et al.

Figure 3. Diagrammatic representation of the functional unit of the ENOX2 proteins which is a dimer, each monomer of which contains two copper centers. During the oxidative portion of the ENOX cycle on the right, the net result is the transfer of 4 electrons plus 4 protons to molecular oxygen to from 2H 2O. The left portion of the diagram illustrates the protein disulfide-thiol interchange activity portion of the cycle where the result is an interchange of protons and electrons that results in the breakage and formation of disulfide bonds important to cell enlargement.

They catalyze both NAD(P)H and hydroquinone The ENOX2 gene is present in the human genome oxidation in one configuration and carry out protein as a single copy, with no obvious homologs and a disulfide-thiol interchange in a second single constitutive ENOX1 (CNOX) ortholog with configuration (Figure 3). The two activities 64% identity and 80% similarity (Jiang et al., alternate creating a regular 22 min period to impart 2008). a time-keeping function (Morré and Morré, 2003). The oscillations are highly synchronous and phased Mutations by low frequency electromagnetic fields. Functionally ENOX2 proteins of cancer cells act as Somatic terminal oxidases of plasma membrane electron No reports of mutations leading to inactivation of or transport (PMET) whereby electrons coming from inability to express ENOX2. cytosolic NAD(P)H are transferred to membrane- located coenzyme Q with eventual transfer of Implicated in electrons and proteins to oxygen to form water (Figure 4). The released energy is presumably Various cancers utilized to drive cell enlargement. The protein Note disulfide-thiol interchange part of the cycle carries The ENOX2 protein is universally associated with out a function essential to the cell enlargement malignancies. It is not the result of an oncogenic mechanism (Morré et al., 2006). The phenotype of mutation but appears to be similar if not identical to unregulated accelerated growth is recapitulated in a a form of ENOX protein with characteristics of an transgenic mouse strain over expressing human oncofetal protein important to maintenance of ENOX2 (Yagiz et al., 2006). unregulated growth in very early development that may be re-expressed in malignancy (Cho and Homology Morré, 2009). Re-expression as an oncofetal protein RNA recognition motif (RRM) in the cell surface helps explain the role of ENOX2 of cancer cells in Ecto-NOX disulfide-thiol exchanger (ECTO-NOX acquiring the well-known characteristic of or ENOX) proteins. This subgroup corresponds to uncontrolled growth. Consistent with this the conserved RNA recognition motif (RRM) in interpretation are observations that the malignant ECTO-NOX proteins (also termed ENOX), phenotypes of invasiveness and growth on soft agar comprising a family of plant and animal NAD(P)H of cancer cells in culture are lost when cells are oxidases exhibiting both oxidative and protein transfected with ENOX2 antisense (Chueh et al., disulfide-like activities. 2004; Tang et al., 2007a).

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 634 ENOX2 (ecto-NOX disulfide-thiol exchanger 2) Tang X, et al.

Figure 4. Schematic representation of the utility of the ENOX2 proteins as cancer-specific cell surface proteins for diagnosis and therapeutic intervention in cancer. Modified from Morré and Morré (2013).

ENOX2 is the first reported cell surface change Breast cancer absent from non-cancer cells and associated with Note most, if not all, forms of human cancer (Morré and Sera of breast cancer patients contains an ENOX2 Morré, 2013). transcript variant of 64 to 69 kDa, isoelectric point As such, ENOX2 emerges as a potential universal 4.2 to 4.9. molecular cancer marker and, being an ecto-protein at the cell surface and shed into the circulation, a Lung cancer reliable cancer diagnostic marker both for cancer Note presence and tissue of cancer origin (Figure 4). Sera of patients with non-small cell lung cancer ENOX2 proteins are expressed differently by contain a 53 to 56 kDa ENOX2 transcript variant, different tissues of cancer origin within the body isoelectric point pH 4.7 to 5.3 while sera of small with each type of cancer being characterized by cell lung cancer contain a transcript variant of 52 one, two, three or more tissue specific transcript kDa, isoelectric point pH 4.1 to 4.6. variants of characteristic molecular weights and isoelectric points (Morré and Morré, 2012). Prostate cancer ENOX2 proteins are absent or reduced to below the Note limits of detection from sera of healthy individuals Sera of patients with prostate cancer contain a 71 to or patients with diseases other than cancer. 88 kDa ENOX2 transcript variant, isoelectric point Circulating ENOX2 has been detected in sera of pH 5.1 to 6.5. patients representing all major forms of human cancer including leukemias and lymphomas. Cervical cancer All ENOX2 transcript variants appear to share the Note common antigenic determinant recognized both by Sera of cervical cancer patients contain a 90 to 100 an ENOX2-specific monoclonal antibody (Cho et kDa transcript variant, isoelectric point pH 4.2 to al., 2002) and a corresponding scFv single chain 5.4. variable region recombinant antibody expressed in Malignant melanoma bacteria and derived from the monoclonal antibody- producing hybridoma cells with analysis by 2-D-gel Note electrophoresis and western blot (Hostetler et al., Sera from malignant melanoma patients contain an 2009). ENOX2 transcript variant of 37 to 41 kDa,

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 635 ENOX2 (ecto-NOX disulfide-thiol exchanger 2) Tang X, et al.

isoelectric point pH 4.6 to 5.3. by anticancer drugs such as doxorubicin, the Leukemias, lymphomas and anticancer sulfonylureas, the vanilloid capsaicin, the catechin EGCg and the cancer isoflavene myelomas phenoxodiol, all of which appear to function as Note quinone site inhibitors directed toward the EEMTE Sera of patients with leukemia, lymphoma or drug binding motif of ENOX2 (Morré and Morré, myeloma, cancers having blood as the common 2013; Hanau et al., 2014). The possibility of tissue of origin, all contain ENOX2 transcript ENOX2 as a drug target is enhanced by the external variants of 38 to 48 kDa and low isoelectric point location of the ENOX2 protein in a position to be pH 3.6 to 4.5. readily available to drugs or antibodies conjugated Ovarian cancer to impermeant supports. As the growth involvement of ENOX2 proteins is in cell enlargement, ENOX2 Note inhibitors also block cell proliferation. The blocked Sera from ovarian carcinoma patients contain two cells, unable to enlarge, also fail to divide and ENOX2 transcript variants of 72 to 90 kDa and 37 eventually undergo apoptosis (Figure 4). to 47 kDa, both having similar isoelectric points in the range of pH 3.7 to 5.0. References Bladder cancer Morré DJ. NADH oxidase activity of HeLa plasma Note membranes inhibited by the antitumor sulfonylurea N-(4- methylphenylsulfonyl)-N'-(4-chlorophenyl) urea Sera of patients with carcinoma of the bladder (LY181984) at an external site. Biochim Biophys Acta. contain two ENOX2 transcript variants of 63 to 66 1995 Dec 13;1240(2):201-8 kDa and 42 to 48 kDa with isoelectric points of 4.2 Kelker M, Kim C, Chueh PJ, Guimont R, Morré DM, Morré to 5.8 and 4.1 to 4.8, respectively. DJ. Cancer isoform of a tumor-associated cell surface NADH oxidase (tNOX) has properties of a prion. Uterine cancer Biochemistry. 2001 Jun 26;40(25):7351-4 Note Cho N, Chueh PJ, Kim C, Caldwell S, Morré DM, Morré Sera of patients with uterine carcinoma contain two DJ. Monoclonal antibody to a cancer-specific and drug- ENOX2 transcript variants of 64 to 69 kDa and 36 responsive hydroquinone (NADH) oxidase from the sera of to 48 kDa with isoelectric points of pH 4.2 to 4.9 cancer patients. Cancer Immunol Immunother. 2002 May;51(3):121-9 and pH 4.5 to 5.6. Chueh PJ, Kim C, Cho N, Morré DM, Morré DJ. Molecular Colorectal cancer cloning and characterization of a tumor-associated, Note growth-related, and time-keeping hydroquinone (NADH) oxidase (tNOX) of the HeLa cell surface. Biochemistry. Sera of patients with colorectal cancer contain at 2002 Mar 19;41(11):3732-41 least two of three possible ENOX2 transcript Morré DJ, Morré DM. Cell surface NADH oxidases (ECTO- variants of 80 to 96 kDa, isoelectric point pH 4.5 to NOX proteins) with roles in cancer, cellular time-keeping, 5.3, 50 to 60 kDa, isoelectric point pH 4.2 to 5.1 growth, aging and neurodegenerative diseases. Free and 33 to 46 kDa, isoelectric point pH 3.8 to 5.2. Radic Res. 2003 Aug;37(8):795-808 Other cancers Chueh PJ, Wu LY, Morré DM, Morré DJ. tNOX is both necessary and sufficient as a cellular target for the Note anticancer actions of capsaicin and the green tea catechin Unique patterns of ENOX2 transcript variant (-)-epigallocatechin-3-gallate. Biofactors. 2004;20(4):235- expression (number, molecular with and isoelectric 49 point) have been found as well associated with Morré DJ, Kim C, Hicks-Berger C. ATP-dependent and brain, endometrial, esophageal, gastric, drug-inhibited vesicle enlargement reconstituted using synthetic lipids and recombinant proteins. Biofactors. hepatocellular renal cell, squamous cell, testicular 2006;28(2):105-17 germ cell and thyroid cancer as well as mesothelioma and sarcomas. Yagiz K, Morré DJ, Morré DM. Transgenic mouse line overexpressing the cancer-specific tNOX protein has an Endometriosis enhanced growth and acquired drug-response phenotype. J Nutr Biochem. 2006 Nov;17(11):750-9 Note Invasive endometriosis is the only non-malignant Tang X, Morré DJ, Morré DM. Antisense experiments demonstrate an exon 4 minus splice variant mRNA as the disorder thus far characterized by the presence of basis for expression of tNOX, a cancer-specific cell surface unique ENOX2 transcript variants. protein. Oncol Res. 2007a;16(12):557-67 As a cancer therapeutic drug target Tang X, Tian Z, Chueh PJ, Chen S, Morré DM, Morré DJ. Alternative splicing as the basis for specific localization of Note tNOX, a unique hydroquinone (NADH) oxidase, to the ENOX2 is responsive to differentiating agents such cancer cell surface. Biochemistry. 2007b Oct as calcitriol and anticancer retinoids and inhibited 30;46(43):12337-46

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Jiang Z, Gorenstein NM, Morré DM, Morré DJ. Molecular Morre DJ, Morre DM.. Early detection: an opportunity for cloning and characterization of a candidate human growth- cancer prevention through early intervention. In: related and time-keeping constitutive cell surface Georgakilas AG (ed), Cancer Prevention. hydroquinone (NADH) oxidase. Biochemistry. 2008 Dec http://www.intechopen.com/download/pdf/35600. In Tech, 30;47(52):14028-38 Rijeka, 2012:389-402 pp. Cho N, Morré DJ. Early developmental expression of a Morre DJ, Morre DM.. ECTO-NOX Proteins. ISBM 978-1- normally tumor-associated and drug-inhibited cell surface- 4614-3957-8. Springer, New York, 2013, 507 pp. located NADH oxidase (ENOX2) in non-cancer cells. Cancer Immunol Immunother. 2009 Apr;58(4):547-52 Hanau C, Morre DJ, Morre DM.. Cancer prevention trial of a synergistic mixture of green tea concentrate plus Hostetler B, Weston N, Kim C, Morre DM, Morre DJ.. Capsicum (CAPSOL-T) in a random population of subjects Cancer site-specific isoforms of ENOX2 (tNOX), a cancer- ages 40-84. specific cell surface oxidase. http://link.springer.com/content/pdf/10.1007%252Fs12014- http://link.springer.com/content/pdf/10.1007%252Fs12014- 008-9016-x. Clin Proteomics 2014: 11(2). 008-9016-x. Clin Proteomics 2009:5:46-51. This article should be referenced as such: Tang X, Kane VD, Morre DM, Morre DJ.. hnRNP F directs formation of an exon 4 minus variant of tumor-associated Tang X, Morré DM, Morré DJ. ENOX2 (ecto-NOX disulfide- NADH oxidase (ENOX2). Mol Cell Biochem. 2011 thiol exchanger 2). Atlas Genet Cytogenet Oncol Nov;357(1-2):55-63. doi: 10.1007/s11010-011-0875-5. Haematol. 2014; 18(9):632-637. Epub 2011 May 28.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 637 Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Gene Section Review

FABP7 (fatty acid binding protein 7, brain) Roseline Godbout, Ho-Yin Poon, Rong-Zong Liu Department of Oncology, University of Alberta, Cross Cancer Institute, 11560 University Avenue, Edmonton, Alberta, T6G 1Z2 Canada (RG, HYP, RZL)

Published in Atlas Database: January 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/FABP7ID46256ch6p22.html DOI: 10.4267/2042/54028 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract Local order: PKIB → FABP7 → SMPDL3A. Review on FABP7, with data on DNA/RNA, on the DNA/RNA protein encoded and where the gene is implicated. Description The FABP7 gene is 4,5 kb long and contains 4 Identity exons, all of which contain coding sequences. The following FABP7 SNPs have been validated: 7 Other names: B-FABP, BLBP, FABPB, MRG in the 3' UTR, 6 in the 5' UTR, 5 missense and 4 HGNC (Hugo): FABP7 SNPs in the coding region that don't alter the amino Location: 6q22.31 acid code (dbSNP).

FABP7 gene. The FABP7 gene is located on chromosome 6 in the region of q22-q23 on the positive strand. Neighboring genes are indicated.

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Crystal structure of FABP7 bound to DHA. The structure of FABP7 is similar to that of other FABPs and consists of two N- terminal α-helices attached to a β-barrel motif (left). Three amino acids are predicted to be important for fatty acid binding: F104 (fuchsia), arginine 126 (red) and Y128 (teal) based on the structure of FABP7 bound to DHA. DHA is shown in yellow (right). Structural data were obtained from the Protein Data Bank (PDB ID: 1FE3) (Balendiran et al., 2000) and rendered using PyMOL (Beaulieu, 2012).

Based on EST data, FABP7 RNA is most highly expressed in the fetus, followed by adult brain, eye, Protein connective tissue, bone, heart, kidney, mammary Description gland, skin, uterus, lung and testis. Transcription regulators: Members of the nuclear FABP7 is a member of the intracellular lipid- factor I (NFI) family regulate the transcription of binding protein family. FABP7 is a 132 amino acid the FABP7 gene (Bisgrove et al., 2000; Brun et al., polypeptide with an estimated molecular mass of 15 2009). kDa. The phosphorylation state of NFI determines its It has a beta-clam structure made up of ten anti- regulatory activity, with FABP7 transcription up- parallel beta sheets capped by two alpha helices. regulated by hypophosphorylated NFI (Bisgrove et Fatty acid ligands reside inside the beta-clam al., 2000; Brun et al., 2013). Other transcription structure. factors implicated in the regulation of FABP7 Expression include Notch (Anthony et al., 2005), PAX6 (Arai FABP7 is expressed in radial glial cells during et al., 2005; Numayama-Tsuruta et al., 2010; Liu et brain development (Feng et al., 1994). FABP7 al., 2012b), and POU-domain protein PBX-1 persists in specific regions of the mature mouse (Josephson et al., 1998). brain, including glia limitans, in radial glial cells of Furthermore, ligands of peroxisome proliferator- the hippocampal dentate gyrus and Bergman glial activated receptors (PPARs) such as clofibrate and cells (Kurtz et al., 1994). FABP7 is also expressed omega-3 docosahexaenoic acid (DHA) have been in glial cells of the peripheral nervous system, and shown to up-regulate FABP7 expression ensheathing cells of the olfactory nerve (Kurtz et (Nasrollahzadeh et al., 2008; Venkatachalam et al., al., 1994). 2012). Post-transcriptional regulation: The 3' untranslated Localisation region of FABP7 contains phylogenetically The FABP7 protein is found in both the cytoplasm conserved cytoplasmic polyadenylation elements and nucleus of normal radial glial cells (Feng et al., (CPE) which have been implicated in the trafficking 1994) and tumor cells (Liang et al., 2006; Slipicevic and localized translation of FABP7 at perisynaptic et al., 2008). FABP7 is also found in perisynaptic processes of astrocytic cells (Gerstner et al., 2012). processes of astrocytes with localized translation of FABP7 at these sites (Gerstner et al., 2012). Pseudogene A predicted FABP7 pseudogene is located on Function chromosome 1 (NCBI nucleotide database Recombinant human FABP7 exhibits the highest NG_029025.1). There are two inferred human affinity for the polyunsaturated omega-3 fatty acids FABP7 pseudogenes listed in the Rat Genome α-linolenic acid, eicosapentaenoic acid, Database docosahexaenoic acid, and for monounsaturated (http://rgd.mcw.edu/rgdweb/report/gene/main.html? omega-9 oleic acid (Kd from 28 to 53 nM) and id=5132511 on chromosome 1 and moderate affinity for the polyunsaturated omega-6 http://www.rgd.mcw.edu/rgdweb/report/gene/main. fatty acids, linoleic acid and arachidonic acid (AA) html?id=6481032 on chromosome 2). (Kd from 115 to 206 nM) (Balendiran et al., 2000).

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 639 FABP7 (fatty acid binding protein 7, brain) Godbout R, et al.

FABP7 has low binding affinity for saturated long increased cell migration (Liang et al., 2005). A role chain fatty acids. Human FABP7 enhances DHA for FABP7 in malignant glioma cell migration was trafficking to the nucleus (Mita et al., 2010). confirmed by Mita et al. (2007) who used human FABP7 is required for the establishment of the U87 malignant glioma cell lines stably transfected radial glial fiber system along which neurons with a FABP7 expression construct to demonstrate migrate in order to reach their correct destination in a correlation between FABP7 expression and the developing brain (Feng et al., 1994). FABP7 is increased cell migration. also required for the maintenance of neuroepithelial In agreement with a role for FABP7 in migration cells in rat cortex (Arai et al., 2005). FABP7 knock- and infiltration, FABP7 was found to be out mice have a structurally normal brain; however, preferentially expressed at sites of infiltration and the mice show enhanced anxiety and increased fear surrounding blood vessels in glioblastoma memory, as well as decreased DHA in neonatal multiforme (Mita et al., 2007). brain and increased AA in adult brain amygdala Growth of malignant glioma cell lines in the (Owada et al., 2006). presence of polyunsaturated fatty acids omega-3 Homology DHA and omega-6 AA indicates that the ratio of AA:DHA affects migration in FABP7-positive Human FABP7 amino acid sequence is 86,4% cells, with a higher DHA:AA ratio resulting in identical to mouse FABP7, 90,9% identical to decreased migration (Mita et al., 2010). These chicken FABP7, 82,6% identical to zebrafish results suggest that glioblastoma tumour growth FABP7a and 78% identical to zebrafish FABP7b. and infiltration may be controlled by increasing Human FABP7 shows variable sequence identity levels of DHA in tumour tissue (Elsherbiny et al., with the other FABP paralogues, with the lowest 2013). identity to FABP1 (27,6%) and highest identity to Neurospheres derived from glioblastoma FABP3 (65,9%). multiforme express high levels of FABP7, suggesting the presence of FABP7-positive neural Mutations stem-like cells in glioblastoma (De Rosa et al., Note 2012). In keeping with this possibility, FABP7 is With the exception of SNPs, no mutations in the preferentially expressed in the subset of FABP7 gene have been reported. glioblastoma tumour cells that express the neural stem cell marker CD133 (Liu et al., 2009). Knock- Implicated in down of FABP7 in glioblastoma-derived Malignant glioma (grades III and IV neurosphere cultures results in decreased cell astrocytoma) / glioblastoma migration and reduced proliferation (De Rosa et al., 2012). multiforme (grade IV astrocytoma) The FABP7 promoter has been shown to be Note hypomethylated in glioblastoma tumours compared FABP7 was first reported to be expressed in to normal brain (Etcheverry et al., 2010). malignant glioma cell lines and malignant glioma tumour tissue in 1998 (Godbout et al., 1998). Liang Breast cancer et al. (2005) used gene expression profiling to Note demonstrate that FABP7 RNA levels were elevated MRG (mammary-derived growth inhibitor-related in glioblastoma tumours compared to normal brain. gene), later shown to be identical to FABP7 These authors showed that elevated levels of (Hohoff and Spener, 1998), was reported to be nuclear FABP7 protein were associated with expressed in normal and benign breast tissue but decreased survival in patients with glioblastoma only rarely in breast cancer (1 of 10 infiltrative multiforme, particularly in younger patients. breast cancers and 2 of 12 ductal carcinomas in Subsequent analysis of 123 glioblastomas by situ) (Shi et al., 1997). Transfection of a MRG Kaloshi et al. (2007) revealed a correlation between expression construct into the MDA-MB-231 breast nuclear FABP7, EGFR amplification and more cancer cell line suppressed cell proliferation and invasive tumours. De Rosa et al. (2012) also tumour growth in an orthotopic mouse model (Shi showed a correlation between elevated FABP7 et al., 1997). Subsequent work showed that MRG levels and decreased survival in patients with over-expression induced differentiation in human glioblastoma multiforme. breast cancer cells and that treatment of breast Transfection of a FABP7 expression construct into cancer cells with DHA causes MRG-dependent the SF767 malignant glioma cell line results in growth inhibition (Wang et al., 2000).

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FABP7 mRNA levels are upregulated in human glioblastoma. Comparison of FABP7 mRNA levels in normal human brain versus glioblastoma tissues. Database obtained from Oncomine website (www.oncomine.org; Bredel Brain 2).

Preferential expression of FABP7 in estrogen ligands available to nuclear receptors such as receptor-negative breast cancer compared to PPARs, understanding the roles of cytoplasmic and estrogen receptor-positive breast cancer has been nuclear FABP7 will help elucidate its biological reported by four separate groups (Tang et al., 2010; functions in breast cancer. Zhang et al., 2010; Graham et al., 2011; Liu et al., 2012a). In an analysis of 176 primary breast Renal cell carcinoma cancers, Liu et al. (2012) found a correlation Note between elevated FABP7 levels and poor FABP7 RNA and protein are up-regulated in renal prognosis. These authors further showed that cell carcinoma compared to normal kidney tissue depletion of FABP7 in FABP7-positive/estrogen- (Seliger et al., 2005; Teratani et al., 2007; Domoto negative MDA-MB-435S, reduced cell growth and et al., 2007). Analysis of a tissue microarray sensitized the cells to growth inhibition by DHA. In containing 272 renal cell carcinomas showed addition, FABP7 was found to mediate DHA- significantly lower levels of FABP7 in grades 3 and induced retinoid-X-receptor beta (RXR β) activation 4 compared to grades 1 and 2 renal cell carcinomas in triple-negative BT-20 breast cancer cells as well (Tölle et al., 2009). No correlation was found as MDA-MB-435S cells. In a study of 899 invasive between patient survival and FABP7 staining breast cancer cases, Zhang et al. (2010) showed that intensity. In agreement with malignant glioma basal breast cancers (estrogen/progesterone experiments, knock-down of FABP7 in human receptor-negative, HER2-negative) that were kidney carcinoma cells resulted in decreased cell FABP7-positive had significantly better outcomes migration (Tölle et al., 2011). than basal breast cancers that were FABP7- The regulation of FABP7 in renal cell carcinoma negative. Analysis of the subcellular localization of has been addressed by analysing the FABP7 FABP7 in 1249 unselected and 245 estrogen promoter (Takaoka et al., 2011). This analysis receptor-negative invasive breast cancers revealed indicates that BRN2 (POU3F2) and nuclear factor I both nuclear and cytoplasmic staining patterns, with (NFI) may be regulating the expression of FABP7 nuclear FABP7 associated with a high histological in renal cell carcinoma. grade, stage, mitotic frequency, as well as basal and triple-negative status (Alshareeda et al., 2012). Melanoma Within the basal category, elevated levels of Note nuclear FABP7 were associated with longer FABP7 been reported to be both down-regulated in disease-free survival. In light of the proposed roles melanoma compared to benign nevi (de Wit et al., for nuclear FABPs in making their fatty acid 2005) and widely expressed in melanoma (Goto et

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al., 2010). FABP7 immunostaining of 149 primary with schizophrenia were up-regulated in the melanomas revealed an association between dorsolateral prefrontal cortex. Furthermore, single FABP7 expression and tumour thickness, as well as nucleotide polymorphism (SNP) analysis revealed a trend towards increased relapse-free survival for an association between missense polymorphism patients who had tumors with low cytoplasmic Thr61Met 182C>T) and male patients with FABP7 levels (Slipicevic et al., 2008). Knock- schizophrenia (Watanabe et al., 2007). down of FABP7 in human melanoma cells resulted In a separate study, FABP7 SNPs F704, F705 and in decreased cell proliferation and invasion. There F709 showed nominal association with bipolar was no association between the nuclear expression disorder (Iwayama et al., 2010). Analysis of 6 of FABP7 and patient survival in this study FABP7 variants identified by polymorphic screen (Slipicevic et al., 2008). failed to identify any associations with autism or Gene expression analysis of 87 primary melanomas schizophrenia in 285 autistic and 1060 and 68 metastatic melanoma, combined with schizophrenic patients of Japanese descent immunohistochemical analysis of 37 paired primary (Maekawa et al., 2010). and metastatic melanomas, showed significantly decreased FABP7 levels in metastatic melanoma References compared to primary tumor tissue (Goto et al., Feng L, Hatten ME, Heintz N. Brain lipid-binding protein 2010). In metastatic melanoma, FABP7 mRNA (BLBP): a novel signaling system in the developing expression was associated with decreased relapse- mammalian CNS. Neuron. 1994 Apr;12(4):895-908 free survival and overall survival (Goto et al., Kurtz A, Zimmer A, Schnütgen F, Brüning G, Spener F, 2010). Loss of heterozygosity analysis using Müller T. The expression pattern of a novel gene encoding microsatellite markers specific to the FABP7 gene brain-fatty acid binding protein correlates with neuronal revealed that 10 of 20 metastatic melanomas (and 0 and glial cell development. Development. 1994 of 14 primary melanomas) had undergone loss of Sep;120(9):2637-49 one FABP7 allele, leading the authors to postulate Shi YE, Ni J, Xiao G, Liu YE, Fuchs A, Yu G, Su J, that genomic instability that favors loss of FABP7 Cosgrove JM, Xing L, Zhang M, Li J, Aggarwal BB, Meager A, Gentz R. Antitumor activity of the novel human expression may lead to better prognosis. breast cancer growth inhibitor, mammary-derived growth Neurological disorders inhibitor-related gene, MRG. Cancer Res. 1997 Aug 1;57(15):3084-91 Note Godbout R, Bisgrove DA, Shkolny D, Day RS 3rd. FABP7 is overexpressed in the brains of Down Correlation of B-FABP and GFAP expression in malignant syndrome patients and has been postulated to glioma. Oncogene. 1998 Apr 16;16(15):1955-62 contribute to Down syndrome-associated Hohoff C, Spener F. Correspondence re: Y.E. Shi et al., neurological disorders (Sánchez-Font et al., 2003). Antitumor activity of the novel human breast cancer growth Pelsers et al. (2004) measured FABP7 levels in inhibitor, mammary-derived growth inhibitor-related gene, various parts of the adult human brain, with a range MRG. Cancer Res., 57: 3084-3091, 1997. Cancer Res. of 0,8 µg/g wet weight in the striatum and 3,1 µg/g 1998 Sep 1;58(17):4015-7 in the frontal lobe. Measurement of FABP7 and Josephson R, Müller T, Pickel J, Okabe S, Reynolds K, FABP3 levels in the serum of patients with minor Turner PA, Zimmer A, McKay RD. POU transcription factors control expression of CNS stem cell-specific genes. brain injuries identified both these FABPs as more Development. 1998 Aug;125(16):3087-100 sensitive at detecting brain injury than markers currently in use for this purpose. Similarly, serum Balendiran GK, Schnutgen F, Scapin G, Borchers T, Xhong N, Lim K, Godbout R, Spener F, Sacchettini JC. FABP7 and FABP3 served as markers for Crystal structure and thermodynamic analysis of human individuals who had undergone ischaemic stroke brain fatty acid-binding protein. J Biol Chem. 2000 Sep (Wunderlich et al., 2005). FABP7 levels were also 1;275(35):27045-54 elevated in the serum of patients with Bisgrove DA, Monckton EA, Packer M, Godbout R. neurodegenerative diseases such as Alzheimer's Regulation of brain fatty acid-binding protein expression by disease, Parkinson's disease and other cognitive differential phosphorylation of nuclear factor I in malignant glioma cell lines. J Biol Chem. 2000 Sep disorders. Although elevated levels of FABP7 were 29;275(39):30668-76 found in only one-third of patients, FABP7 is still the most discriminatory serum marker identified to Wang M, Liu YE, Ni J, Aygun B, Goldberg ID, Shi YE. Induction of mammary differentiation by mammary-derived date (Teunissen et al., 2011). The authors propose growth inhibitor-related gene that interacts with an omega- that elevated levels of FABP7 in serum may reflect 3 fatty acid on growth inhibition of breast cancer cells. damage to the central nervous system. Cancer Res. 2000 Nov 15;60(22):6482-7 FABP7-deficient mice have characteristics Sánchez-Font MF, Bosch-Comas A, Gonzàlez-Duarte R, associated with schizophrenia such as decreased Marfany G. Overexpression of FABP7 in Down syndrome prepulse inhibition and shortened startle response fetal brains is associated with PKNOX1 gene-dosage latency (Watanabe et al., 2007). FABP7 RNA imbalance. Nucleic Acids Res. 2003 Jun 1;31(11):2769-77 levels in the postmortem brains of male patients

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Nuclear FABP7 immunoreactivity is preferentially expressed in infiltrative Goto Y, Koyanagi K, Narita N, Kawakami Y, Takata M, glioma and is associated with poor prognosis in EGFR- Uchiyama A, Nguyen L, Nguyen T, Ye X, Morton DL, Hoon overexpressing glioblastoma. BMC Cancer. 2006 Apr DS. Aberrant fatty acid-binding protein-7 gene expression 19;6:97 in cutaneous malignant melanoma. J Invest Dermatol. 2010 Jan;130(1):221-9 Owada Y, Abdelwahab SA, Kitanaka N, Sakagami H, Takano H, Sugitani Y, Sugawara M, Kawashima H, Kiso Y, Iwayama Y, Hattori E, Maekawa M, Yamada K, Toyota T, Mobarakeh JI, Yanai K, Kaneko K, Sasaki H, Kato H, Ohnishi T, Iwata Y, Tsuchiya KJ, Sugihara G, Kikuchi M, Saino-Saito S, Matsumoto N, Akaike N, Noda T, Kondo H. Hashimoto K, Iyo M, Inada T, Kunugi H, Ozaki N, Iwata N, Altered emotional behavioral responses in mice lacking Nanko S, Iwamoto K, Okazaki Y, Kato T, Yoshikawa T. brain-type fatty acid-binding protein gene. Eur J Neurosci. Association analyses between brain-expressed fatty-acid 2006 Jul;24(1):175-87 binding protein (FABP) genes and schizophrenia and bipolar disorder. Am J Med Genet B Neuropsychiatr Domoto T, Miyama Y, Suzuki H, Teratani T, Arai K, Genet. 2010 Mar 5;153B(2):484-93 Sugiyama T, Takayama T, Mugiya S, Ozono S, Nozawa R. Evaluation of S100A10, annexin II and B-FABP expression Maekawa M, Iwayama Y, Arai R, Nakamura K, Ohnishi T, as markers for renal cell carcinoma. Cancer Sci. 2007 Toyota T, Tsujii M, Okazaki Y, Osumi N, Owada Y, Mori N, Jan;98(1):77-82 Yoshikawa T. Polymorphism screening of brain-expressed FABP7, 5 and 3 genes and association studies in autism Kaloshi G, Mokhtari K, Carpentier C, Taillibert S, Lejeune and schizophrenia in Japanese subjects. J Hum Genet. J, Marie Y, Delattre JY, Godbout R, Sanson M. FABP7 2010 Feb;55(2):127-30 expression in glioblastomas: relation to prognosis, invasion and EGFR status. J Neurooncol. 2007 Sep;84(3):245-8 Mita R, Beaulieu MJ, Field C, Godbout R. Brain fatty acid- binding protein and omega-3/omega-6 fatty acids: Mita R, Coles JE, Glubrecht DD, Sung R, Sun X, Godbout mechanistic insight into malignant glioma cell migration. J R. B-FABP-expressing radial glial cells: the malignant Biol Chem. 2010 Nov 19;285(47):37005-15 glioma cell of origin? Neoplasia. 2007 Sep;9(9):734-44 Numayama-Tsuruta K, Arai Y, Takahashi M, Sasaki- Teratani T, Domoto T, Kuriki K, Kageyama T, Takayama T, Hoshino M, Funatsu N, Nakamura S, Osumi N. Ishikawa A, Ozono S, Nozawa R. Detection of transcript Downstream genes of Pax6 revealed by comprehensive for brain-type fatty Acid-binding protein in tumor and urine transcriptome profiling in the developing rat hindbrain. of patients with renal cell carcinoma. Urology. 2007 BMC Dev Biol. 2010 Jan 18;10:6 Feb;69(2):236-40

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 643 FABP7 (fatty acid binding protein 7, brain) Godbout R, et al.

Tang XY, Umemura S, Tsukamoto H, Kumaki N, Tokuda Y, Osamura RY. Overexpression of fatty acid binding protein-7 correlates with basal-like subtype of breast 10.1371/journal.pone.0052113. Epub 2012 Dec 21. cancer. Pathol Res Pract. 2010 Feb 15;206(2):98-101 Gerstner JR, Vanderheyden WM, LaVaute T, Westmark Zhang H, Rakha EA, Ball GR, Spiteri I, Aleskandarany M, CJ, Rouhana L, Pack AI, Wickens M, Landry CF.. Time of Paish EC, Powe DG, Macmillan RD, Caldas C, Ellis IO, day regulates subcellular trafficking, tripartite synaptic Green AR. The proteins FABP7 and OATP2 are localization, and polyadenylation of the astrocytic Fabp7 associated with the basal phenotype and patient outcome mRNA. J Neurosci. 2012 Jan 25;32(4):1383-94. doi: in human breast cancer. Breast Cancer Res Treat. 2010 10.1523/JNEUROSCI.3228-11.2012. May;121(1):41-51 Liu RZ, Graham K, Glubrecht DD, Lai R, Mackey JR, Graham K, Ge X, de Las Morenas A, Tripathi A, Godbout R.. A fatty acid-binding protein 7/RXRb pathway Rosenberg CL. Gene expression profiles of estrogen enhances survival and proliferation in triple-negative breast receptor-positive and estrogen receptor-negative breast cancer. J Pathol. 2012a Nov;228(3):310-21. doi: cancers are detectable in histologically normal breast 10.1002/path.4001. Epub 2012 Apr 18. epithelium. Clin Cancer Res. 2011 Jan 15;17(2):236-46 Liu RZ, Monckton EA, Godbout R.. Regulation of the Takaoka N, Takayama T, Teratani T, Sugiyama T, Mugiya FABP7 gene by PAX6 in malignant glioma cells. Biochem S, Ozono S. Analysis of the regulation of fatty acid binding Biophys Res Commun. 2012b Jun 8;422(3):482-7. doi: protein 7 expression in human renal carcinoma cell lines. 10.1016/j.bbrc.2012.05.019. Epub 2012 May 11. BMC Mol Biol. 2011 Jul 19;12:31 Venkatachalam AB, Lall SP, Denovan-Wright EM, Wright Teunissen CE, Veerhuis R, De Vente J, Verhey FR, JM.. Tissue-specific differential induction of duplicated fatty Vreeling F, van Boxtel MP, Glatz JF, Pelsers MA. Brain- acid-binding protein genes by the peroxisome proliferator, specific fatty acid-binding protein is elevated in serum of clofibrate, in zebrafish (Danio rerio). BMC Evol Biol. 2012 patients with dementia-related diseases. Eur J Neurol. Jul 9;12:112. doi: 10.1186/1471-2148-12-112. 2011 Jun;18(6):865-71 Brun M, Glubrecht DD, Baksh S, Godbout R.. Calcineurin Tölle A, Krause H, Miller K, Jung K, Stephan C. regulates nuclear factor I dephosphorylation and activity in Importance of brain type fatty acid binding protein for cell- malignant glioma cell lines. J Biol Chem. 2013 Aug biological processes in human renal carcinoma cells. 16;288(33):24104-15. doi: 10.1074/jbc.M113.455832. Oncol Rep. 2011 May;25(5):1307-12 Epub 2013 Jul 9. Alshareeda AT, Rakha EA, Nolan CC, Ellis IO, Green AR. Elsherbiny ME, Emara M, Godbout R.. Interaction of brain Fatty acid binding protein 7 expression and its sub-cellular fatty acid-binding protein with the polyunsaturated fatty localization in breast cancer. Breast Cancer Res Treat. acid environment as a potential determinant of poor 2012 Jul;134(2):519-29 prognosis in malignant glioma. Prog Lipid Res. 2013 Oct;52(4):562-70. doi: 10.1016/j.plipres.2013.08.004. Epub Beaulieu M.. Mechanistic Insight Into the Role of FABP7 in 2013 Aug 24. Malignant Glioma. Ann Arbor, 2012 University of Alberta (Canada) This article should be referenced as such: De Rosa A, Pellegatta S, Rossi M, Tunici P, Magnoni L, Godbout R, Poon HY, Liu RZ. FABP7 (fatty acid binding Speranza MC, Malusa F, Miragliotta V, Mori E, Finocchiaro protein 7, brain). Atlas Genet Cytogenet Oncol Haematol. G, Bakker A.. A radial glia gene marker, fatty acid binding 2014; 18(9):638-644. protein 7 (FABP7), is involved in proliferation and invasion of glioblastoma cells. PLoS One. 2012;7(12):e52113. doi:

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Gene Section Review

GSTA1 (glutathione S-transferase alpha 1) Ana Savic-Radojevic, Tanja Radic Institute of Medical and Clinical Biochemistry, Faculty of Medicine, University of Belgrade, Serbia (ASR, TR)

Published in Atlas Database: January 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/GSTA1ID40764ch6p12.html DOI: 10.4267/2042/54029 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

genes (GSTA1, GSTA2, GSTA3, GSTA4, GSTA5) Abstract and seven pseudogenes (Morel et al., 2002). Review on GSTA1, with data on DNA/RNA, on the protein encoded and where the gene is implicated. Description The GSTA1 gene is approximately 12 kb in length Identity and is closely flanked by other alpha class gene sequences. The complete sequence of the 1,7-kb Other names: GST2, GSTA1-1, GTH1 intergenic region between exon 7 of an upstream HGNC (Hugo): GSTA1 pseudogene and exon 1 of the GSTA1 gene has Location: 6p12.2 been determined (Suzuki et al., 1993). Local order Transcription Between the LOC647169 (similar to glutathione The 1276-nucleotide transcript encodes a protein of transferase) and GSTA6P (glutathione S-transferase 222 amino acid residues. alpha 6 pseudogene) (according to PubMed). Pseudogene Note The GSTA1 gene is composed of 7 exons spanning An additional gene that encodes an uncharacterized a region of 12487 bases. Alpha class GST has been identified. The protein derived from this gene would have 19 amino acid DNA/RNA substitutions compared with the GSTA1 isoenzyme. Several pseudogenes with single-base and/or Note complete exon deletions have been identified, but The human alpha class genes are located in a cluster no reverse-transcribed pseudogenes have been on chromosome 6p12 and comprise five functional detected (Suzuki et al., 1993).

GSTA1 gene. The GSTA1 gene spans a region of 12,5 kb composed of the seven exons (red) and six introns (green). Exons 1, 2, 3, 4, 5, 6 and 7 are 59 bp, 117 bp, 52 bp, 133 bp, 142 bp, 132 bp and 198 bp in length, respectively.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 645 GSTA1 (glutathione S-transferase alpha 1) Savic-Radojevic A, Radic T

Crystal structure of human glutathione transferase (GST) A1-1 in complex with glutathione. Adapted from PDB (Grahn et al., 2006).

Polymorphisms: GSTA1 has a functional three little effect on GSTA1 expression (Morel et al., apparently linked single nucleotide polymorphisms 2002). (SNPs) in an SP1-responsive element within the proximal promoter (G-52A, C-69T and T-567G), Protein plus at least four SNPs further upstream and a silent SNP A-375G. Two variants, GSTA1*A (-567T, - Note 69C,-52G) and GSTA1*B (-67G, -69T, -52A), have Glutathione S-transferase A1 is N-terminally been named according to the linked functional processed. SNPs. Specifically, these substitutions result in Amino acids: 222. differential expression with lower transcriptional Calculated molecular mass: 25,63 kDa. activation of variant GSTA1*B than common GSTA1*A allele. It has been suggested that this Description genetic variation can change an individual's The active GSTA1-1 enzyme is a homodimer, with susceptibility to carcinogens and toxins, as well as, each subunit containing a GSH-binding site (G-site) affect the efficacy of some drugs (Coles and and a second adjacent hydrophobic binding site for Kadlubar, 2003). In addition, the linkage the electrophilic substrate (H-site) (Wilce and disequilibrium between GSTA1*A/GSTA1*B and Parker, 1994). GSTA2G335C (Ser112Thr) has been shown in The C-terminal region of GSTA1-1 contributes to Caucasians: specifically, GSTA1*A/GSTA2C335 the catalytic and noncatalytic ligand-binding (Thr112) and GSTA1*B/GSTA2G335 (Ser112) functions of the enzyme, while the conserved G-site (Ning et al., 2004). It seems that the higher hepatic is located in the N-terminal domain (Balogh et al., expression of GSTA1 enzyme in homozygous 2009). GSTA1 individuals is associated with the lower Protein flexibility and dynamics in a molten hepatic expression of GSTA2 in GSTA2C335 globule-active site including the C-terminal α9 (Thr112) individuals (Coles et al., 2001a; Ning et helix and the protruding ends of the α4-α5 helices al., 2004). Other haplotypes within this result in achieving remarkable catalytic promiscuity nomenclature but including SNPs C-115T, T-631G, of GSTA1-1 (Wu and Dong, 2012; Honaker et al., and C-1142G also have been proposed 2013). It has been proposed that the α9 helix may (Bredschneider et al., 2002; Guy et al., 2004). function as a mobile gate to the active-site cavity, Polymorphisms upstream of G-52C seem to have controlling substrate access and product release.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 646 GSTA1 (glutathione S-transferase alpha 1) Savic-Radojevic A, Radic T

Structure determination and refinement of human alpha class glutathione transferase A1-1, and a comparison with the MU and PI class enzymes. Adapted from PDB (Sinning et al., 1993).

Expression Function GSTA1-1 is highly expressed (as mRNA and Human GSTA1-1 enzyme catalyzes the GSH- protein) in liver, intestine, kidney, adrenal gland, dependent detoxification of electrophiles showing pancreas and testis, while expression in a wide highly promiscuous substrate selectivity for many range of tissues is low (Hayes and Pulford, 1995; structurally unrelated chemicals, including Coles et al., 2001a). Both positive and negative environmental carcinogens (e.g. benzo(a)pyrene regulatory regions are present in the 5` noncoding diol epoxides), several alkylating chemotherapeutic region of GSTA1, including a polymorphic SP1- agents (such as busulfan, chlorambucil, melphalan, binding site within the proximal promoter. Binding phosphoramide mustard, cyclophosphamide, of the transcription factor AP1 has been suggested thiotepa), as well as, steroids and products of lipid as a common mechanism for up-regulation of GSTs degradation. GSTA1-1 is the most highly expressed (Hayes and Pulford, 1995). The results of recent GST of the liver and could therefore, be critical for study also implied the role of a Kelch-like ECH- "systemic" detoxification of electrophilic associated protein 1 (Keap1)-dependent signaling xenobiotics including carcinogens and drugs (Coles pathway for the induction of the constitutive and Kadlubar, 2005). GSTA1 expression during epithelial cell In addition to enzymatic detoxification, GSTA1 differentiation (Kusano et al., 2008). Regarding 5 4 acts as modulator of mitogen-activated protein GSH-dependent ∆ -∆ isomerase activity of this kinase (MAPK) signal transduction pathway via a class of enzyme, it has been shown that mechanism involving protein-protein interactions. steroidogenic factor 1 (SF-1) is involved in Namely, GSTA1 forms complexes with c-Jun N- regulation of expression of GSTA genes terminal kinase (JNK), modifying JNK activation (Matsumura et al., 2013). Aberrant overexpression during cellular stress (Adnan et al., 2012). has been observed in various malignancies such as Thus, it is possible that GSTA1 confer drug colorectal (Hengstler et al., 1998) and lung cancer resistance by two distinct means: by direct (Carmichael et al., 1988), while decrease in alpha inactivation (detoxification) of chemotherapeutic class GSTs has been observed in stomach and liver drugs and by acting as inhibitors of MAPK tumors (Howie et al., 1990). A detailed recent pathway. review on GSTA1 can be found in Wu and Dong, 2012. Homology Localisation The alpha class GSTs is showing strong intra-class sequence similarity (Balogh et al., 2009). Cytosolic.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 647 GSTA1 (glutathione S-transferase alpha 1) Savic-Radojevic A, Radic T

from acute myeloid leukemia patients, showing Mutations resistance to doxorubicin in vitro (Sargent et al., Germinal 1999). In addition, GSTA1 and CYP39A1 (member of cytochrome P450 family) polymorphisms were None described so far. found to be associated with pharmacokinetics of Somatic busulfan, which is used in preparative regimens 36 mutations (COSMIC): 26 substitution-missense, prior to stem cell transplantation in pediatric 4 substitution-nonsense, 5 substitution-coding patients (ten Brink et al., 2013). silent, 1 unknown type. Prostate cancer Note Implicated in Genetic variants of GSTA1 and GSTT1 may Colorectal cancer modify prostate cancer risk, especially among smokers (Komiya et al., 2005). Note Regarding the role of GSTA1 polymorphism in the Asthma risk of colorectal cancer, the results of Note epidemiological studies are still inconclusive. Genetic alterations in GST enzymes may influence Several studies showed that GSTA1*B genotype the detoxification of environmental toxic (low hepatic expression) is associated with substances in airway and increase the risk of increased susceptibility to colorectal cancer, which asthma. imply the possible inefficient hepatic detoxification Thus, it has been shown that subjects with at least of food-derived carcinogen metabolite N-acetoxy- one allele -69T in the GSTA1 genotype have an PhIP (Coles et al., 2001b; Sweeney et al., 2002). In increased risk of asthma (Polimanti et al., 2010). contrast, meta-analysis of Economopoulos representing the pooled analysis of four studies References (1648 cases, 2039 controls) does not confer this association. Carmichael J, Forrester LM, Lewis AD, Hayes JD, Hayes PC, Wolf CR. Glutathione S-transferase isoenzymes and Breast cancer glutathione peroxidase activity in normal and tumour samples from human lung. Carcinogenesis. 1988 Note Sep;9(9):1617-21 The role of GSTA1 polymorphism in breast cancer Howie AF, Forrester LM, Glancey MJ, Schlager JJ, Powis risk was mainly based on investigation on response G, Beckett GJ, Hayes JD, Wolf CR. Glutathione S- to chemotherapeutic drugs in these patients. In transferase and glutathione peroxidase expression in breast cancer patients on cyclophosphamide normal and tumour human tissues. Carcinogenesis. 1990 containing chemotherapy carriers of GSTA1*B/*B Mar;11(3):451-8 genotype showed significantly reduced five years Sinning I, Kleywegt GJ, Cowan SW, Reinemer P, Dirr HW, risk of death in comparison to GSTA1*A Huber R, Gilliland GL, Armstrong RN, Ji X, Board PG. Structure determination and refinement of human alpha homozygous carriers. This association was likely class glutathione transferase A1-1, and a comparison with caused by decreased detoxification of the the Mu and Pi class enzymes. J Mol Biol. 1993 Jul therapeutic metabolites of cyclophosphamide in 5;232(1):192-212 GSTA1*B/*B patients (Sweeney et al., 2003). Suzuki T, Johnston PN, Board PG. Structure and organization of the human alpha class glutathione S- Bladder cancer transferase genes and related pseudogenes. Genomics. Note 1993 Dec;18(3):680-6 Recent investigation indicates that the GSTA1-low Wilce MC, Parker MW. Structure and function of activity genotype in combination with the GSTM1- glutathione S-transferases. Biochim Biophys Acta. 1994 null genotype significantly increases the risk of Mar 16;1205(1):1-18 bladder cancer in smokers (Matic et al., 2013). In Hayes JD, Pulford DJ. The glutathione S-transferase addition, it seems that GSTA1 polymorphism may supergene family: regulation of GST and the contribution influence vulnerability to oxidative DNA damage, of the isoenzymes to cancer chemoprotection and drug resistance. Crit Rev Biochem Mol Biol. 1995;30(6):445-600 thereby contributing to the malignant potential of transitional cell carcinoma (Savic-Radojevic et al., Hengstler JG, Böttger T, Tanner B, Dietrich B, Henrich M, Knapstein PG, Junginger T, Oesch F. Resistance factors 2013). in colon cancer tissue and the adjacent normal colon Myeloid leukemia tissue: glutathione S-transferases alpha and pi, glutathione and aldehyde dehydrogenase. Cancer Lett. 1998 Jun Note 5;128(1):105-12 Aberant overexpression of both GSTA1 and Sargent JM, Williamson C, Hall AG, Elgie AW, Taylor CG. GSTA2 proteins was found in blast cells derived Evidence for the involvement of the glutathione pathway in

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 648 GSTA1 (glutathione S-transferase alpha 1) Savic-Radojevic A, Radic T

drug resistance in AML. Adv Exp Med Biol. 1999;457:205- Biol Crystallogr. 2006 Feb;62(Pt 2):197-207 9 Kusano Y, Horie S, Shibata T, Satsu H, Shimizu M, Hitomi Coles BF, Morel F, Rauch C, Huber WW, Yang M, Teitel E, Nishida M, Kurose H, Itoh K, Kobayashi A, Yamamoto CH, Green B, Lang NP, Kadlubar FF. Effect of M, Uchida K. Keap1 regulates the constitutive expression polymorphism in the human glutathione S-transferase A1 of GST A1 during differentiation of Caco-2 cells. promoter on hepatic GSTA1 and GSTA2 expression. Biochemistry. 2008 Jun 10;47(23):6169-77 Pharmacogenetics. 2001a Nov;11(8):663-9 Balogh LM, Le Trong I, Kripps KA, Tars K, Stenkamp RE, Coles B, Nowell SA, MacLeod SL, Sweeney C, Lang NP, Mannervik B, Atkins WM. Structural analysis of a Kadlubar FF. The role of human glutathione S-transferases glutathione transferase A1-1 mutant tailored for high (hGSTs) in the detoxification of the food-derived catalytic efficiency with toxic alkenals. Biochemistry. 2009 carcinogen metabolite N-acetoxy-PhIP, and the effect of a Aug 18;48(32):7698-704 polymorphism in hGSTA1 on colorectal cancer risk. Mutat Res. 2001b Oct 1;482(1-2):3-10 Economopoulos KP, Sergentanis TN. GSTM1, GSTT1, GSTP1, GSTA1 and colorectal cancer risk: a Bredschneider M, Klein K, Mürdter TE, Marx C, comprehensive meta-analysis. Eur J Cancer. 2010 Eichelbaum M, Nüssler AK, Neuhaus P, Zanger UM, Jun;46(9):1617-31 Schwab M. Genetic polymorphisms of glutathione S- transferase A1, the major glutathione S-transferase in Polimanti R, Piacentini S, Moscatelli B, Pellicciotti L, human liver: consequences for enzyme expression and Manfellotto D, Fuciarelli M. GSTA1, GSTO1 and GSTO2 busulfan conjugation. Clin Pharmacol Ther. 2002 gene polymorphisms in Italian asthma patients. Clin Exp Jun;71(6):479-87 Pharmacol Physiol. 2010 Aug;37(8):870-2 Morel F, Rauch C, Coles B, Le Ferrec E, Guillouzo A. The Adnan H, Antenos M, Kirby GM. The effect of menadione human glutathione transferase alpha locus: genomic on glutathione S-transferase A1 (GSTA1): c-Jun N- organization of the gene cluster and functional terminal kinase (JNK) complex dissociation in human characterization of the genetic polymorphism in the colonic adenocarcinoma Caco-2 cells. Toxicol Lett. 2012 hGSTA1 promoter. Pharmacogenetics. 2002 Oct 2;214(1):53-62 Jun;12(4):277-86 Wu B, Dong D. Human cytosolic glutathione transferases: Sweeney C, Coles BF, Nowell S, Lang NP, Kadlubar FF. structure, function, and drug discovery. Trends Pharmacol Novel markers of susceptibility to carcinogens in diet: Sci. 2012 Dec;33(12):656-68 associations with colorectal cancer. Toxicology. 2002 Dec Honaker MT, Acchione M, Zhang W, Mannervik B, Atkins 27;181-182:83-7 WM. Enzymatic detoxication, conformational selection, and Coles BF, Kadlubar FF. Detoxification of electrophilic the role of molten globule active sites. J Biol Chem. 2013 compounds by glutathione S-transferase catalysis: Jun 21;288(25):18599-611 determinants of individual response to chemical Matic M, Pekmezovic T, Djukic T, Mimic-Oka J, Dragicevic carcinogens and chemotherapeutic drugs? Biofactors. D, Krivic B, Suvakov S, Savic-Radojevic A, Pljesa- 2003;17(1-4):115-30 Ercegovac M, Tulic C, Coric V, Simic T. GSTA1, GSTM1, Sweeney C, Ambrosone CB, Joseph L, Stone A, Hutchins GSTP1, and GSTT1 polymorphisms and susceptibility to LF, Kadlubar FF, Coles BF. Association between a smoking-related bladder cancer: a case-control study. Urol glutathione S-transferase A1 promoter polymorphism and Oncol. 2013 Oct;31(7):1184-92 survival after breast cancer treatment. Int J Cancer. 2003 Matsumura T, Imamichi Y, Mizutani T, Ju Y, Yazawa T, Mar 1;103(6):810-4 Kawabe S, Kanno M, Ayabe T, Katsumata N, Fukami M, Guy CA, Hoogendoorn B, Smith SK, Coleman S, Inatani M, Akagi Y, Umezawa A, Ogata T, Miyamoto K. O'Donovan MC, Buckland PR. Promoter polymorphisms in Human glutathione S-transferase A (GSTA) family genes glutathione-S-transferase genes affect transcription. are regulated by steroidogenic factor 1 (SF-1) and are Pharmacogenetics. 2004 Jan;14(1):45-51 involved in steroidogenesis. FASEB J. 2013 Aug;27(8):3198-208 Ning B, Wang C, Morel F, Nowell S, Ratnasinghe DL, Carter W, Kadlubar FF, Coles B. Human glutathione S- Savic-Radojevic A, Djukic T, Simic T, Pljesa-Ercegovac M, transferase A2 polymorphisms: variant expression, Dragicevic D, Pekmezovic T, Cekerevac M, Santric V, distribution in prostate cancer cases/controls and a novel Matic M. GSTM1-null and GSTA1-low activity genotypes form. Pharmacogenetics. 2004 Jan;14(1):35-44 are associated with enhanced oxidative damage in bladder cancer. Redox Rep. 2013;18(1):1-7 Coles BF, Kadlubar FF. Human alpha class glutathione S- transferases: genetic polymorphism, expression, and ten Brink MH, van Bavel T, Swen JJ, van der Straaten T, susceptibility to disease. Methods Enzymol. 2005;401:9-42 Bredius RG, Lankester AC, Zwaveling J, Guchelaar HJ. Effect of genetic variants GSTA1 and CYP39A1 and age Komiya Y, Tsukino H, Nakao H, Kuroda Y, Imai H, Katoh on busulfan clearance in pediatric patients undergoing T. Human glutathione S-transferase A1, T1, M1, and P1 hematopoietic stem cell transplantation. polymorphisms and susceptibility to prostate cancer in the Pharmacogenomics. 2013 Nov;14(14):1683-90 Japanese population. J Cancer Res Clin Oncol. 2005 Apr;131(4):238-42 This article should be referenced as such: Grahn E, Novotny M, Jakobsson E, Gustafsson A, Grehn Savic-Radojevic A, Radic T. GSTA1 (glutathione S- L, Olin B, Madsen D, Wahlberg M, Mannervik B, Kleywegt transferase alpha 1). Atlas Genet Cytogenet Oncol GJ. New crystal structures of human glutathione Haematol. 2014; 18(9):645-649. transferase A1-1 shed light on glutathione binding and the conformation of the C-terminal helix. Acta Crystallogr D

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 649 Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Gene Section Short Communication

MOAP1 (Modulator Of Apoptosis 1) Gamze Ayaz, Mesut Muyan Department of Biological Sciences, Middle East Technical University, Ankara, Turkey (GA, MM)

Published in Atlas Database: January 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/MOAP1ID46494ch14q32.html DOI: 10.4267/2042/54030 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract Protein Short communication on MOAP1, with data on Note DNA/RNA, on the protein encoded and where the MOAP1 is a short-lived protein with a half-life of gene is implicated. 25 minutes. It is degraded by the ubiquitin-proteosome system Identity (Fu et al., 2007). Other names: MAP-1, PNMA4 Description HGNC (Hugo): MOAP1 MOAP-1 is a 351 amino-acid long protein with a Location: 14q32.12 molecular mass of 39.5 kDa. Isoelectric point (pI) of MOAP-1 is 4.939 at pH 7.0. DNA/RNA MOAP-1 contains a BH3 (Bcl-2 homology 3) like domain required for homodimerization and Description interaction with Bcl-2 associated X (Bax) protein. Under normal condition, MOAP1 is held as an Human MOAP1 gene contains three exons and the inactive conformation through intramolecular encoding sequence of 1056 bases is in the exon 3. interactions. Interaction between RASSF1A (ras- Transcription association domain family 1, isoform A) and MOAP1 is a 351 amino-acid long protein. MOAP1 reduces the inhibitory intramolecular interaction of MOAP1 and allows MOAP1 through Pseudogene the BH3 like domain to bind Bax (Baksh et al., No reported pseudogenes. 2005).

The MOAP1 gene is located in the reverse strand. Exons are shown as boxes and introns as lines. The filled box in the exon 3 is the coding sequence of MOAP1. Numbers below represent the size of exon or intron in base pair.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 650 MOAP1 (Modulator Of Apoptosis 1) Ayaz G, Muyan M

Expression the downregulation of MOAP1 expression and the aggressiveness of breast cancer (Law, 2012). MOAP1 is expressed in the adipose, adrenal, blood, brain, breast, colon and heart. MOAP1 is expressed in higher level in the heart and brain. To be noted Localisation Note miRNA: It is reported that miR-1228 prevents MOAP-1 protein localizes in the cytoplasm (Law, cellular apoptosis by binding to the 3'UTR of 2012) and also seen in the mitochondria (Tan et al., MOAP1 mRNA, thereby decreasing MOAP-1 2005). protein levels (Yan and Zhao, 2012). Function MOAP1 is a BH3-like protein that acts as a pro- References apoptotic molecule (Tan et al., 2001). When Tan KO, Tan KM, Chan SL, Yee KS, Bevort M, Ang KC, overexpressed, MOAP1 induces caspase-dependent Yu VC. MAP-1, a novel proapoptotic protein containing a apoptosis in mammalian cells (Tan et al., 2005). BH3-like motif that associates with Bax through its Bcl-2 Studies showed that TNF α stimulation recruits homology domains. J Biol Chem. 2001 Jan 26;276(4):2802-7 RASSF1A and MOAP1 to death receptors complexes. This recruitment leads to the Baksh S, Tommasi S, Fenton S, Yu VC, Martins LM, association of RASSF1A with MOAP1 and to the Pfeifer GP, Latif F, Downward J, Neel BG. The tumor suppressor RASSF1A and MAP-1 link death receptor induction of a conformational change in MOAP1. signaling to Bax conformational change and cell death. Mol This results in the opening of the BH3 domain to Cell. 2005 Jun 10;18(6):637-50 allow MOAP1 to interact with Bax. Bax is Tan KO, Fu NY, Sukumaran SK, Chan SL, Kang JH, Poon subsequently inserted into the mitochondrial KL, Chen BS, Yu VC. MAP-1 is a mitochondrial effector of membrane leading to apoptosis (Baksh et al., 2005; Bax. Proc Natl Acad Sci U S A. 2005 Oct Foley et al., 2008). 11;102(41):14623-8 Homology Fu NY, Sukumaran SK, Yu VC. Inhibition of ubiquitin- mediated degradation of MOAP-1 by apoptotic stimuli Also highly conserved in rat (Rattus norvegicus) promotes Bax function in mitochondria. Proc Natl Acad Sci and mouse (Mus musculus), human MOAP1 U S A. 2007 Jun 12;104(24):10051-6 protein shares 99% amino acids identity with that of Foley CJ, Freedman H, Choo SL, Onyskiw C, Fu NY, Yu chimpanzee (Pan troglodytes) (Law et al., 2012). VC, Tuszynski J, Pratt JC, Baksh S. Dynamics of RASSF1A/MOAP-1 association with death receptors. Mol Mutations Cell Biol. 2008 Jul;28(14):4520-35 Law J.. MOAP1: A Candidate Tumor Suppressor Protein Note Master Thesis, University of Alberta, Canada, 2012 Gene mutations have not been described yet. http://hdl.handle.net/10402/era.24828. Law J, Yu VC, Baksh S.. Modulator of Apoptosis 1: A Implicated in Highly Regulated RASSF1A-Interacting BH3-Like Protein. Mol Biol Int. 2012;2012:536802. doi: Breast cancer 10.1155/2012/536802. Epub 2012 Jun 14. Disease Yan B, Zhao JL.. miR-1228 prevents cellular apoptosis through targeting of MOAP1 protein. Apoptosis. 2012 Microarray analysis of 176 primary, treatment- Jul;17(7):717-24. doi: 10.1007/s10495-012-0710-9. naive breast cancer and of 10 normal breast tissue samples suggests that the MOAP1 gene expression This article should be referenced as such: is significantly down-regulated in breast cancer. It Ayaz G, Muyan M. MOAP1 (Modulator Of Apoptosis 1). appears that there is a positive correlation between Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):650- 651.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 651 Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Gene Section Short Communication

PHLDA1 (pleckstrin homology-like domain, family A, member 1) Maria Aparecida Nagai Discipline of Oncology, Department of Radiology and Oncology, Medical School, University of Sao Paulo, Center for Translational Investigation in Oncology, Cancer Institute from Sao Paulo State, Sao Paulo, Brazil (MAN)

Published in Atlas Database: January 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/PHLDA1ID41707ch12q15.html DOI: 10.4267/2042/54031 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

PHLDA1 protein has a modular structure Abstract containing a central pleckstrin homology-like Short communication on PHLDA1, with data on domain (PHL) and prolin-glutamine (PQ) and DNA/RNA, on the protein encoded and where the proline-histidine (PH) repeats in the C-terminal gene is implicated. region (see figure above). Identity Expression PHLDA1 is widely expressed in mammalian tissues Other names: DT1P1B11, PHRIP, TDAG51 displaying cytoplasmic, vesicle membrane, plasma HGNC (Hugo): PHLDA1 membrane and nuclear subcellular localization. Location: 12q21.2 PHLDA1 expression is up-regulated by estrogen, IGF-1 (insulin-like growth factor 1), FGF Local order: Minus strand. (fibroblast growth factor), TPA (phorbol ester), and ER (endoplamic reticulum)-stress agents such as DNA/RNA homocysteine, tunicamicyne, and farnesol. Description Localisation PHLDA1 gene contains 2 exons, 1 intervening Cytoplasm, vesicle membrane, plasma membrane, sequence and spans 6,3 kb of genomic DNA. nucleus. Transcription Function 1,2 kb mRNA. Protein binding. Several evidences have implicate Pseudogene PHLDA1 as a potential transcriptional activator that acts as a pro-apoptotic and antiproliferative factor, Not identified. however the mechanisms by which PHLDA1 Protein mediates cell survival is still under investigation. Homology Description PHLDA2 (pleckstrin homology-like domain, family The PHLDA1 gene encodes for a 401 amino acid A, member 2) and PHLDA3 (pleckstrin homology- protein that is a member of the evolutionarily like domain, family A, member 3) are paralogs for conserved pleckstrin homology-like domain family. PHLDA1.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 652 PHLDA1 (pleckstrin homology-like domain, family A, member 1) Nagai MA

Schematic representation of the modular structure of PHLDA1 protein. PHL: pleckstrin homology-like domain spanning amino acids residues from 150 to 283; QQ: proline/glutamine rich sequence (aa residues from 189 to 204); PQ: proline- glutamine tracts (aa residues from 311 to 346); PH: proline-histidine-rich tracts (aa residues from 352 to 389); *: indicates phosphorylation sites.

significantly better in patients with tumors that Mutations were negative for PHLDA1, and a multivariate Note analysis suggested that PHLDA1 is an independent Short genetic variation - dbSNP: rs139162669, prognostic factor in OSCC patients (Coutinho- rs73385441, rs74620794, rs147230079, Camillo et al., 2013). rs76437300, rs186978611, rs140610935, Colon cancer rs144470255, rs79545253, rs147644129. Note Implicated in Altered PHLDA1 expression has been shown to be associated with the process of intestinal Melanoma tumorigenesis (Sakthianandeswaren et al., 2011). Note Basal cell carcinoma PHLDA1 expression was associated with reduced Note cell growth and colony formation and with PHLDA1 has also been shown to be a follicular and increased apoptotic rates and drug sensitivity in epithelial stem cell marker (Ohyama et al., 2006; melanoma cell lines. Loss of PHLDA1 has been Sakthianandeswaren et al., 2011) with potential to correlated with melanoma progression (Neef et al., differentiates between trichoepithelioma and basal 2002). cell carcinoma (Sellheyer and Nelson, 2011). Breast cancer Atherosclerosis Note Note Down-regulation of PHLDA1 mRNA and protein In vivo and in vitro studies demonstrated that expression is frequently observed in primary increased PHLDA1 expression induced by invasive breast tumours. homocysteine promotes detachment-mediated Down-regulation of PHLDA1 protein has been programmed cell death and contributes to the shown to be a strong predictor of poor prognosis for development of atherosclerosis (Hossain et al., breast cancer patients, indicating that reduced 2003). PHLDA1 expression contribute for breast cancer Genetic variant in an intergenic region of the progression and might serve as useful prognostic PHLDA1 gene (rs2367446) has been shown to be biomarker of disease outcome (Nagai et al., 2007). associated with the development of cardiovascular Oral cancer diseases (Hossain et al., 2013). Note Epilepsy Reduced expression of PHLDA1 was observed in Note 60,7% of oral squamous cell carcinomas (OSCC), PHLDA1 expression has been shown to be higher especially in well-differentiated tumors. in the anterior temporal neocortex from patients Positive PHLDA1 immunostaining was associated with intractable epilepsy when compared with the with advanced clinical stages of the disease, levels observed in the neocortex from the control suggesting that PHLDA1 has a functional role in group, suggesting a possible association of oral tumorigenesis. PHLDA1 in the physiopathology of the disease (Xi Overall and disease-free survival rates were et al., 2007).

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 653 PHLDA1 (pleckstrin homology-like domain, family A, member 1)Nagai MA

neocortex of patients with intractable epilepsy. Neurosci References Lett. 2007 Sep 20;425(1):53-8 Park CG, Lee SY, Kandala G, Lee SY, Choi Y. A novel Marchiori AC, Casolari DA, Nagai MA. Transcriptional up- gene product that couples TCR signaling to Fas(CD95) regulation of PHLDA1 by 17beta-estradiol in MCF-7 breast expression in activation-induced cell death. Immunity. cancer cells. Braz J Med Biol Res. 2008 Jul;41(7):579-82 1996 Jun;4(6):583-91 Johnson EO, Chang KH, de Pablo Y, Ghosh S, Mehta R, Frank D, Mendelsohn CL, Ciccone E, Svensson K, Badve S, Shah K. PHLDA1 is a crucial negative regulator Ohlsson R, Tycko B. A novel pleckstrin homology-related and effector of Aurora A kinase in breast cancer. J Cell gene family defined by Ipl/Tssc3, TDAG51, and Tih1: Sci. 2011 Aug 15;124(Pt 16):2711-22 tissue-specific expression, chromosomal location, and parental imprinting. Mamm Genome. 1999 Sakthianandeswaren A, Christie M, D'Andreti C, Tsui C, Dec;10(12):1150-9 Jorissen RN, Li S, Fleming NI, Gibbs P, Lipton L, Malaterre J, Ramsay RG, Phesse TJ, Ernst M, Jeffery RE, Poulsom Kuske MD, Johnson JP. Assignment of the human R, Leedham SJ, Segditsas S, Tomlinson IP, Bernhard OK, PHLDA1 gene to chromosome 12q15 by radiation hybrid Simpson RJ, Walker F, Faux MC, Church N, Catimel B, mapping. Cytogenet Cell Genet. 2000;89(1-2):1 Flanagan DJ, Vincan E, Sieber OM. PHLDA1 expression marks the putative epithelial stem cells and contributes to Neef R, Kuske MA, Pröls E, Johnson JP. Identification of intestinal tumorigenesis. Cancer Res. 2011 May the human PHLDA1/TDAG51 gene: down-regulation in 15;71(10):3709-19 metastatic melanoma contributes to apoptosis resistance and growth deregulation. Cancer Res. 2002 Oct Sellheyer K, Nelson P. Follicular stem cell marker PHLDA1 15;62(20):5920-9 (TDAG51) is superior to cytokeratin-20 in differentiating between trichoepithelioma and basal cell carcinoma in Hossain GS, van Thienen JV, Werstuck GH, Zhou J, Sood small biopsy specimens. J Cutan Pathol. 2011 SK, Dickhout JG, de Koning AB, Tang D, Wu D, Falk E, Jul;38(7):542-50 Poddar R, Jacobsen DW, Zhang K, Kaufman RJ, Austin RC. TDAG51 is induced by homocysteine, promotes Nagai MA.. PHLDA1 (pleckstrin homology-like domain, detachment-mediated programmed cell death, and family A, member). Encyclopedia of Signaling Molecules contributes to the cevelopment of atherosclerosis in 2012, pp 1365 - 1369. ISBN: 978-4419-04060-7 (Editor: hyperhomocysteinemia. J Biol Chem. 2003 Aug Sandun Choi) 8;278(32):30317-27 Coutinho-Camillo CM, Lourenco SV, Nonogaki S, Oberg HH, Sipos B, Kalthoff H, Janssen O, Kabelitz D. Vartanian JG, Nagai MA, Kowalski LP, Soares FA.. Regulation of T-cell death-associated gene 51 (TDAG51) Expression of PAR-4 and PHLDA1 is prognostic for overall expression in human T-cells. Cell Death Differ. 2004 and disease-free survival in oral squamous cell Jun;11(6):674-84 carcinomas. Virchows Arch. 2013 Jul;463(1):31-9. doi: 10.1007/s00428-013-1438-9. Epub 2013 Jun 9. Toyoshima Y, Karas M, Yakar S, Dupont J, Lee Helman, LeRoith D. TDAG51 mediates the effects of insulin-like Hossain GS, Lynn EG, Maclean KN, Zhou J, Dickhout JG, growth factor I (IGF-I) on cell survival. J Biol Chem. 2004 Lhotak S, Trigatti B, Capone J, Rho J, Tang D, McCulloch Jun 11;279(24):25898-904 CA, Al-Bondokji I, Malloy MJ, Pullinger CR, Kane JP, Li Y, Shiffman D, Austin RC.. Deficiency of TDAG51 protects Ohyama M, Terunuma A, Tock CL, Radonovich MF, Pise- against atherosclerosis by modulating apoptosis, Masison CA, Hopping SB, Brady JN, Udey MC, Vogel JC. cholesterol efflux, and peroxiredoxin-1 expression. J Am Characterization and isolation of stem cell-enriched human Heart Assoc. 2013 May 17;2(3):e000134. doi: hair follicle bulge cells. J Clin Invest. 2006 Jan;116(1):249- 10.1161/JAHA.113.000134. 60 Nagai MA, Fregnani JH, Netto MM, Brentani MM, Soares This article should be referenced as such: FA. Down-regulation of PHLDA1 gene expression is Nagai MA. PHLDA1 (pleckstrin homology-like domain, associated with breast cancer progression. Breast Cancer family A, member 1). Atlas Genet Cytogenet Oncol Res Treat. 2007 Nov;106(1):49-56 Haematol. 2014; 18(9):652-654. Xi ZQ, Wang LY, Sun JJ, Liu XZ, Zhu X, Xiao F, Guan LF, Li JM, Wang L, Wang XF. TDAG51 in the anterior temporal

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 654 Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Gene Section Review

ADAMTS15 (ADAM Metallopeptidase With Thrombospondin Type 1 Motif, 15) Santiago Cal, Alvaro J Obaya Departamento de Bioquimica y Biologia Molecular, Instituto Universitario de Oncologia (IUOPA), Universidad de Oviedo, 33006, Asturias, Spain (SC), Biologia Funcional, Instituto Universitario de Oncologia (IUOPA), Universidad de Oviedo, 33006, Asturias, Spain (AJO)

Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/ADAMTS15ID45587ch11q24.html DOI: 10.4267/2042/54032 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

protein, with an estimated molecular weight of Abstract 103,2 kDa. ADAMTS-15 shares a structural Review on ADAMTS15, with data on DNA/RNA, multidomain complex architecture with the rest of on the protein encoded and where the gene is the members of the ADAMTS family. implicated. This organization includes a signal peptide, a prodomain involved in maintaining enzyme latency Identity and a catalytic domain that contains the consensus sequence HEXXHGXXHD involved in the HGNC (Hugo): ADAMTS15 coordination of the zinc atom necessary for Location: 11q24.3 catalytic activiy of the enzyme. This sequence ends in an Asp residue which DNA/RNA distinguishes ADAMTSs from other metalloproteases such as MMPs. Following this Description catalytic region there are several other domains 8 exons, spans approximately 27.66 Kb of genomic characterized as disintegrin-like domain, a central DNA in the centromere-to-telomere orientation. thrombospondin-1 (TSP-1) motif, a cysteine-rich The translation initiation codon is located to exon 1, domain, a spacer region and two more TSP-1 and the stop codon to exon 8. domains (Cal et al., 2002). Protein Expression ADAMTS15 cDNA was originally cloned from Description both, a human liver and kidney fetal cDNA library The open reading fame encodes a 950 amino acid (Cal et al., 2002).

Domain organization of ADAMTS-15. Pro: prodomain; TSP: thrombospondin type-1 domains.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 655 ADAMTS15 (ADAM Metallopeptidase With Thrombospondin Type 1 Motif, 15) Cal S, Obaya AJ

Later on, in the search for proteinases and chicken (Refseq: XM_417874), and zebrafish proteinase inhibitors in articular cartilage from (Refseq: XM_001341842). femoral heads of patients with end-stage osteoarthritis (OA) Kevorkian et al. found high Mutations levels of ADAMTS-15 expression in samples from both, OA patients as well as normal controls Somatic (Kevorkian et al., 2004). In relation with ADAMTS15 was identified as one of the so-called ADAMTS-15 participation in tumor progression its CAN genes found to be mutated in a small set of expression has been described in either normal cells colorectal cancers (Sjöblom et al., 2006). Two or cells adjacent or marginal to cancer tissue in heterozigous somatic mutations were described out samples from colon adenocarcinoma as well as in of eleven human cancer samples (cDNA: samples from head and neck squamous cell 2309A>G, cDNA: 2632T>G). Functional relevance carcinoma (Viloria et al., 2009; Stokes et al., 2010). of mutations found in colorectal cancer were Additionally ADAMTS-15 presence has also been described for a deleterious single base mutation detected in some breast and prostate cancer cell 24544 ∆G affecting the two carboxy-terminal lines (Molokwu et al., 2010). thrombospondin motifs of ADAMTS-15 (Viloria et Localisation al., 2009). The derived truncated form of ADAMTS-15 (ADAMTS15_G849fs) is barely Extracellular, mostly pericellular. found in the pericellular space of the cell being Function mostly liberated to the culture media. Functional Few studies describe ADAMTS-15 function studies revealed ADAMTS15_G849fs not showing beyond those describing its participation in cancer the anti-tumoral properties of full length and osteoartritic processes. Regarding cancer, ADAMTS-15. In the same study, three other ADAMTS-15 has recently emerged as a putative mutations where identified, a base pair mutation tumor suppresor gene since it is downregulated in affecting the second TSP-1 domain (24616C>T), a breast cancer, and functionally inactivated through silent base pair change (13777C>T) and another specific mutations in colorectal cancer (Porter et al., base deletion generating a completely truncated 2004; Porter et al., 2006; Viloria et al., 2009). In form of ADAMTS-15 (366 ∆) (Viloria et al., 2009). addition, aberrant expression of ADAMTS-15 is implicated in prostate cancer progression Implicated in (Molokwu et al., 2010). The latest apparently Various cancers results from the relationship between ADAMTS-15 expression and versican degradation. Thus, Note ADAMTS-15 seems to be acting as a versican- ADAMTS-15 has recently emerged as a putative degrading enzyme whose accumulation potentially tumor suppresor gene since it is downregulated in contributes to prostate cancer pathology (Cross et breast cancer, and functionally inactivated through al., 2005). In this regard, versican seems to be one specific mutations in colorectal cancer (Porter et al., of the targets of ADAMTS-15 proteolityc activity 2006; López-Otín et al., 2009; Viloria et al., 2009). which involves this protein in processes such as In addition, aberrant expression of ADAMTS-15 is cancer or skeletal muscle fiber formation (Croos et implicated in prostate cancer progression (Cross et al., 2005; Stupka et al., 2013; Dancevic et al., al., 2005; Molokwu et al., 2010). The first 2013). indication regarding a potential protective role for ADAMTS15 derived from the observation that low Homology ADAMTS15 expression levels coupled to high ADAMTS-15 belongs to the A Disintegrin And ADAMTS8 levels conferred poor prognosis to Metalloprotease Domains with ThromboSpondin breast cancer patients (Porter et al., 2006). motifs (ADAMTS) family, which consists of 19 Moreover, ADAMTS15 was identified as one of the secreted zinc metalloproteinases (Porter et al., so-called CAN genes found to be mutated in a 2005). All members of the family share the same small set of colorectal cancers (Sjöblom et al., structural domain design. ADAMTS-15 is, among 2006). Functional support to the putative relevance all the members, closely related to ADAMTS-1 of ADAMTS-15 as a tumor suppresor protease was which suggested its involvement in angiogenic described after finding four additional mutations in processes (Cal et al., 2002). ADAMTS-15 gene sequence in human colon The ADAMTS15 gene is conserved in chimpanzee carcinomas (Viloria et al., 2009). Two of the new (Refseq: XM_522253), macaque (Refseq: mutations resulted in the generation of truncated XM_001113698), dog (Refseq: XM_005620295), forms of ADAMTS-15, one of them lacking the last cow (Refseq: NM_001192390), mouse (Refseq: two thrombospondin domains whereas the other NM_001024139), rat (Refseq: NM_001106810), originating a complete ADAMTS-15 knock-down.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 656 ADAMTS15 (ADAM Metallopeptidase With Thrombospondin Type 1 Motif, 15) Cal S, Obaya AJ

Functional analysis revealed that the presence of the Breast cancer two last thrombospondin domains is important for Note the pericellular loacalization of ADAMTS-15 and ADAMTS15 elevated expression correlates with affects the anti-tumoral function of full length favorable outcome in patients with breast cancer ADAMTS-15 (Viloria et al., 2009; Dancevic et al., (Porter et al., 2006). 2013). More recently, ADAMTS-15 has been described as a head and neck squamous cell carcinoma References (HNSCC)-associated proteinase since its expression Ricciardelli C, Mayne K, Sykes PJ, Raymond WA, McCaul is elevated (together with ADAMTS-1 and K, Marshall VR, Horsfall DJ. Elevated levels of versican ADAMTS-8) in areas surrounding HNSCC tumor but not decorin predict disease progression in early-stage prostate cancer. Clin Cancer Res. 1998 Apr;4(4):963-71 microenvironment (Demircan et al., 2009; Stokes et al., 2010). Cal S, Obaya AJ, Llamazares M, Garabaya C, Quesada V, López-Otín C. Cloning, expression analysis, and structural In addition, these three members of the ADAMTS characterization of seven novel human ADAMTSs, a family family have elevated expression levels in HNSCC of metalloproteinases with disintegrin and thrombospondin- tumor versus normal tissue and in HNSCC derived 1 domains. Gene. 2002 Jan 23;283(1-2):49-62 cell lines vs normal keratinocytes (Stokes et al., Luo J, Dunn T, Ewing C, Sauvageot J, Chen Y, Trent J, 2010). Isaacs W. Gene expression signature of benign prostatic ADAMTS-15 has also been indirectly involved in hyperplasia revealed by cDNA microarray analysis. androgen-mediated prostate cancer growth and Prostate. 2002 May 15;51(3):189-200 proliferation, function that depends on ADAMTS- Kevorkian L, Young DA, Darrah C, Donell ST, Shepstone 15 versicanolytic activity (Cross et al., 2005; L, Porter S, Brockbank SM, Edwards DR, Parker AE, Clark Molokwu et al., 2010). IM. Expression profiling of metalloproteinases and their inhibitors in cartilage. Arthritis Rheum. 2004 Jan;50(1):131- Molokwu et al identified one androgen-responsive 41 element (ARE) in ADAMTS-15 promoter and 12 Porter S, Scott SD, Sassoon EM, Williams MR, Jones JL, more AREs in its gene sequence. In the same article Girling AC, Ball RY, Edwards DR. Dysregulated the authors demonstrated ADAMTS-15 reduction expression of adamalysin-thrombospondin genes in both, at mRNA and protein levels, in the presence human breast carcinoma. Clin Cancer Res. 2004 Apr of dihidrotestorone (DHT). 1;10(7):2429-40 ADAMTS-15 down-regulation in prostate cancer Cross NA, Chandrasekharan S, Jokonya N, Fowles A, resulted in high versican levels which is a poor Hamdy FC, Buttle DJ, Eaton CL. The expression and prognosis indicator in these type of tumors regulation of ADAMTS-1, -4, -5, -9, and -15, and TIMP-3 by TGFbeta1 in prostate cells: relevance to the (Ricciardelli et al., 1998; Luo et al., 2002; accumulation of versican. Prostate. 2005 May Molokwu et al., 2010). 15;63(3):269-75 Colon cancer Porter S, Clark IM, Kevorkian L, Edwards DR. The ADAMTS metalloproteinases. Biochem J. 2005 Feb Note 15;386(Pt 1):15-27 ADAMTS15 expression inversely correlates with Porter S, Span PN, Sweep FC, Tjan-Heijnen VC, histopathologic differentiation grade in human Pennington CJ, Pedersen TX, Johnsen M, Lund LR, colorectal carcinomas when analyzing ADAMTS- Rømer J, Edwards DR. ADAMTS8 and ADAMTS15 15 inmunostaining in normal colon epithelia, well- expression predicts survival in human breast carcinoma. differentiated tumors, moderately differentiated Int J Cancer. 2006 Mar 1;118(5):1241-7 tumors, and poorly differentiated colorectal Sjöblom T, Jones S, Wood LD, Parsons DW, Lin J, Barber carcinomas (Viloria et al., 2009). TD, Mandelker D, Leary RJ, Ptak J, Silliman N, Szabo S, Buckhaults P, Farrell C, Meeh P, Markowitz SD, Willis J, Head and neck squamous carcinoma Dawson D, Willson JK, Gazdar AF, Hartigan J, Wu L, Liu C, Parmigiani G, Park BH, Bachman KE, Papadopoulos N, (HNSCC) Vogelstein B, Kinzler KW, Velculescu VE. The consensus Note coding sequences of human breast and colorectal cancers. ADAMTS15 mRNA levels, together with those of Science. 2006 Oct 13;314(5797):268-74 other ADAMTS members (ADAMTS1, Demircan K, Gunduz E, Gunduz M, Beder LB, Hirohata S, ADAMTS4, ADAMTS5, ADAMTS8, Nagatsuka H, Cengiz B, Cilek MZ, Yamanaka N, Shimizu K, Ninomiya Y. Increased mRNA expression of ADAMTS ADAMTS9), were reduced in HNSCC primary metalloproteinases in metastatic foci of head and neck tumors compared with paired non-cancerous tissues cancer. Head Neck. 2009 Jun;31(6):793-801 (Demircan et al., 2009). Regarding tumor López-Otín C, Palavalli LH, Samuels Y. Protective roles of microenvironment ADAMTS15 expression is matrix metalloproteinases: from mouse models to human elevated in adjacent and margin tissue when cancer. Cell Cycle. 2009 Nov 15;8(22):3657-62 compared with tumor center tissue (Stokes et al., Viloria CG, Obaya AJ, Moncada-Pazos A, Llamazares M, 2010). Astudillo A, Capellá G, Cal S, López-Otín C. Genetic

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 657 ADAMTS15 (ADAM Metallopeptidase With Thrombospondin Type 1 Motif, 15) Cal S, Obaya AJ

inactivation of ADAMTS15 metalloprotease in human disintegrin-like and metalloproteinase domain with colorectal cancer. Cancer Res. 2009 Jun 1;69(11):4926-34 thrombospondin-1 repeats-15: a novel versican-cleaving proteoglycanase. J Biol Chem. 2013 Dec Molokwu CN, Adeniji OO, Chandrasekharan S, Hamdy FC, 27;288(52):37267-76 Buttle DJ. Androgen regulates ADAMTS15 gene expression in prostate cancer cells. Cancer Invest. 2010 Stupka N, Kintakas C, White JD, Fraser FW, Hanciu M, Aug;28(7):698-710 Aramaki-Hattori N, Martin S, Coles C, Collier F, Ward AC, Apte SS, McCulloch DR. Versican processing by a Stokes A, Joutsa J, Ala-Aho R, Pitchers M, Pennington CJ, disintegrin-like and metalloproteinase domain with Martin C, Premachandra DJ, Okada Y, Peltonen J, thrombospondin-1 repeats proteinases-5 and -15 facilitates Grénman R, James HA, Edwards DR, Kähäri VM. myoblast fusion. J Biol Chem. 2013 Jan 18;288(3):1907-17 Expression profiles and clinical correlations of degradome components in the tumor microenvironment of head and This article should be referenced as such: neck squamous cell carcinoma. Clin Cancer Res. 2010 Apr 1;16(7):2022-35 Cal S, Obaya AJ. ADAMTS15 (ADAM Metallopeptidase With Thrombospondin Type 1 Motif, 15). Atlas Genet Dancevic CM, Fraser FW, Smith AD, Stupka N, Ward AC, Cytogenet Oncol Haematol. 2014; 18(9):655-658. McCulloch DR. Biosynthesis and expression of a

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Gene Section Review

ADRB2 (adrenoceptor beta 2, surface) Denise Tostes Oliveira, Diego Mauricio Bravo-Calderón Department of Stomatology, Area of Pathology, Bauru School of Dentistry - University of Sao Paulo, Bauru, Brazil (DTO, DMBC)

Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/ADRB2ID43818ch5q32.html DOI: 10.4267/2042/54033 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

2006). The receptor is comprised of 413 amino acid Abstract residues of approximately 46500 daltons (Johnson, Review on ADRB2, with data on DNA/RNA, on 2006). β2 adrenergic receptor is N-glycosylated at the protein encoded and where the gene is amino acids 6, 15, and 187; these are important for implicated. roper insertion of the receptor into the membrane as well as for agonist trafficking (McGraw and Identity Liggett, 2005; Johnson, 2006). Other names: ADRB2R, ADRBR, B2AR, BAR, Expression BETA2AR β2 adrenergic receptor is widely distributed, this HGNC (Hugo): ADRB2 protein is expressed by airway smooth muscle (30- Location: 5q32 40000 per cell), epithelial and endothelial cells of the lung, smooth muscle of blood vessels, skeletal DNA/RNA muscle, mast cells, lymphocytes, oral and skin keratinocytes and also by diverse cancer cells Description (Kohm and Sanders, 2001; Lutgendorf et al., 2003; ADBR2 gene spans about 2,04 kb and consists of Johnson, 2006; Sood et al., 2006; Thaker et al., one exon. 2006; Yang et al., 2006; Sastry et al., 2007; Yu et al., 2007; Liu et al., 2008a; Liu et al., 2008b; Shang Transcription et al., 2009; Sivamani et al., 2009; Yang et al., ADBR2 no has introns in either their coding or 2009; Bernabé et al., 2011; Bravo-Calderón et al., untranslated sequences. The primary transcripts are 2011-2012; Steenhuis et al., 2011; Zhang et al., processed at their 5' and 3' ends like other 2011; Loenneke et al., 2012). premessenger RNAs, but no splicing is needed. Localisation Pseudogene β2 adrenergic receptor is a transmembrane protein. No pseudogenes have been reported. Like all GPCRs, the β2 adrenergic receptor has seven transmembrane a domains that form a pocket Protein containing binding sites for agonists and competitive antagonists (McGraw and Liggett, Description 2005; Johnson, 2006). There are 3 extracellular β2 adrenergic receptor is a member of the loops, with one being the amino terminus, and 3 superfamily of G-protein coupled receptors intracellular loops, with a carboxy terminus (GPCRs) (McGraw and Liggett, 2005; Johnson, (McGraw and Liggett, 2005; Johnson, 2006).

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Activation of protein kinase A (PKA) by signal transduction of β2 adrenergic receptor (adapted of Rosenbaum et al., 2009).

Function foradrenergic receptors in these experimental effects. Agonist binding of β2 adrenergic receptor results in Norepinephrine was later found to increase the in activation of Gs protein. The Gs protein a subunit vitro invasive potential of ovarian cancer cells, an stimulates adenylyl cyclase to generate cyclic 3'-5'- effect that was blocked by propranolol (Sood et al., adenosine monophosphate (cAMP), which in 2006). Norepinephrine also increased tumor cell sequence activates the cAMP-dependent protein expression of matrix metalloproteinase-2 (MMP-2) kinase A (PKA) and the agonist-occupied receptor and MMP-9, and pharmacologic blockade of is phosphorylated. MMPs abrogated the effects of norepinephrine on After phosphorylation, the receptor switches its tumor cell invasive potential (Sood et al., 2006). coupling specificity to Gi. GTP-bound Gi α In the same way, Thaker et al. (Thaker et al., 2006) dissociates from the heterodimeric G βγ , and free correlated chronic behavioral stress with higher Gβγ subunits mediate activation of the MAP kinase levels of tissue catecholamines and more invasive signaling pathway in the same way as Gi-coupled growth of ovarian carcinoma cells in an orthotopic receptors. Increase of intracellular cAMP levels mouse model. These effects were mediated through leads diverse cell functions as cell proliferation, β2 adrenergic receptor activation of PKA signaling differentiation, angiogenesis and migration (Daaka pathway (Thaker et al., 2006). Tumors in stressed et al., 1997). animals showed increased vascularization and enhanced expression of VEGF, MMP2 and MMP9; Implicated in these effects could be abrogated by propranolol Ovarian carcinoma (Thaker et al., 2006). Note Prostate cancer Reverse transcriptase-PCR studies indicated Note constitutive expression of β2 adrenergic receptor on β2 adrenergic receptor signaling was related to ovarian carcinoma cell lines (Lutgendorf et al., prostate cancer cell progression (Sastry et al., 2007; 2003). Lutgendorf et al. (Lutgendorf et al., 2003) Zhang et al., 2011). β2 adrenergic receptor investigated the effects of norepinephrine and activation of PKA signaling pathway has been isoproterenol (a nonspecific-adrenergic agonist) on associated with reduction of sensitivity of prostate the production of vascular endothelial growth factor cancer cells to apoptosis (Sastry et al., 2007) and (VEGF) by ovarian cancer cell lines; and found that promotion of cell proliferation and cell migration both, norepinephrine and isoproterenol, (Zhang at al., 2011). significantly enhanced VEGF production. These Contrastingly, other investigation demonstrated that effects were blocked by thenon-specific β the genetic silencing of β2 adrenergic receptor antagonist propranolol, supporting a role increases cell migration and invasion of normal

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prostate cells and that the weak expression of this carcinoma cells lines (Yang et al., 2006); as well protein is associated with metastases and with worst upregulated the production of VEGF, interleukin survival rates in prostate cancer patients (Yu et al., (IL)-8, and IL-6 in human melanoma tumor cell 2007). lines (Yang et al., 2009). Esophageal squamous cell References carcinoma Daaka Y, Luttrell LM, Lefkowitz RJ. Switching of the Note coupling of the beta2-adrenergic receptor to different G Liu et al. (Liu et al., 2008b) demonstrated that proteins by protein kinase A. Nature. 1997 Nov stimulation of β2 adrenergic receptor with 6;390(6655):88-91 epinephrine significantly increase the esophageal Kohm AP, Sanders VM. Norepinephrine and beta 2- cancer cell proliferation accompanied by elevation adrenergic receptor stimulation regulate CD4+ T and B of the expression of VEGF, VEGF receptor lymphocyte function in vitro and in vivo. Pharmacol Rev. VEGFR-1 and VEGFR-2. In addition, it has been 2001 Dec;53(4):487-525 shown that the epidermal growth factor mediates Lutgendorf SK, Cole S, Costanzo E, Bradley S, Coffin J, the mitogenic signals in esophageal cancer cells Jabbari S, Rainwater K, Ritchie JM, Yang M, Sood AK. Stress-related mediators stimulate vascular endothelial through transactivation of β2 adrenergic receptor growth factor secretion by two ovarian cancer cell lines. (Liu et al., 2008a). Clin Cancer Res. 2003 Oct 1;9(12):4514-21 Oral squamous cell carcinoma McGraw DW, Liggett SB. Molecular mechanisms of beta2- adrenergic receptor function and regulation. Proc Am (OSCC) Thorac Soc. 2005;2(4):292-6; discussion 311-2 Note Johnson M. Molecular mechanisms of beta(2)-adrenergic Genetic and protein expression of β2 adrenergic receptor function, response, and regulation. J Allergy Clin receptor was demonstrated in OSCC by using RT- Immunol. 2006 Jan;117(1):18-24; quiz 25 PCR assay, Western blot and Sood AK, Bhatty R, Kamat AA, Landen CN, Han L, Thaker immunohistochemistry (Shang et al., 2009; Bernabé PH, Li Y, Gershenson DM, Lutgendorf S, Cole SW. Stress et al., 2011; Bravo-Calderón et al., 2011-2012). hormone-mediated invasion of ovarian cancer cells. Clin Investigations performed in different oral cancer Cancer Res. 2006 Jan 15;12(2):369-75 cell lines demonstrated that β2 adrenergic receptor Thaker PH, Han LY, Kamat AA, Arevalo JM, Takahashi R, signaling by norepinephrine increases cell Lu C, Jennings NB, Armaiz-Pena G, Bankson JA, Ravoori proliferation and invasion, and upregulates M, Merritt WM, Lin YG, Mangala LS, Kim TJ, Coleman RL, Landen CN, Li Y, Felix E, Sanguino AM, Newman RA, interleukin-6 (IL-6) gene expression and protein Lloyd M, Gershenson DM, Kundra V, Lopez-Berestein G, release (Shang et al., 2009; Bernabé et al., 2011). Lutgendorf SK, Cole SW, Sood AK. Chronic stress Furthermore, Shang et al. (Shang et al., 2009) promotes tumor growth and angiogenesis in a mouse reported that malignant cell positive model of ovarian carcinoma. Nat Med. 2006 Aug;12(8):939-44 immunoexpression of β2-AR was significantly correlated with age, tumor size, clinical stage and Yang EV, Sood AK, Chen M, Li Y, Eubank TD, Marsh CB, Jewell S, Flavahan NA, Morrison C, Yeh PE, Lemeshow S, cervical lymph node metastasis in OSCC patients, Glaser R. Norepinephrine up-regulates the expression of and that β2-AR may play an important role in the vascular endothelial growth factor, matrix formation and metastasis of oral cancer. However, a metalloproteinase (MMP)-2, and MMP-9 in retrospective clinical study of a large number of nasopharyngeal carcinoma tumor cells. Cancer Res. 2006 patients showed that patients with OSCC who Nov 1;66(21):10357-64 exhibited strong β2-AR immunohistochemical Sastry KS, Karpova Y, Prokopovich S, Smith AJ, Essau B, expression by malignant epithelial cells Gersappe A, Carson JP, Weber MJ, Register TC, Chen YQ, Penn RB, Kulik G. Epinephrine protects cancer cells demonstrated higher survival rates compared to from apoptosis via activation of cAMP-dependent protein patients with weak/negative β2-AR expression kinase and BAD phosphorylation. J Biol Chem. 2007 May (Bravo-Calderón et al., 2011-2012). Therefore, 11;282(19):14094-100 further clinical and laboratory studies are warranted Yu J, Cao Q, Mehra R, Laxman B, Yu J, Tomlins SA, to elucidate the role of β2 adrenergic receptor Creighton CJ, Dhanasekaran SM, Shen R, Chen G, Morris activation in oral squamous cell carcinoma. DS, Marquez VE, Shah RB, Ghosh D, Varambally S, Chinnaiyan AM. Integrative genomics analysis reveals Various cancers silencing of beta-adrenergic signaling by polycomb in prostate cancer. Cancer Cell. 2007 Nov;12(5):419-31 Note β2 adrenergic receptor was also Liu X, Wu WK, Yu L, Li ZJ, Sung JJ, Zhang ST, Cho CH. Epidermal growth factor-induced esophageal cancer cell immunohistochemically identified in proliferation requires transactivation of beta- nasopharyngeal carcinoma (Yang et al., 2006) and adrenoceptors. J Pharmacol Exp Ther. 2008a in melanoma (Yang et al., 2009). Norepinephrine Jul;326(1):69-75 treatment increased MMP-2, MMP-9, and VEGF Liu X, Wu WK, Yu L, Sung JJ, Srivastava G, Zhang ST, levels in culture supernatants of nasopharyngeal Cho CH. Epinephrine stimulates esophageal squamous-

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cell carcinoma cell proliferation via beta-adrenoceptor- carcinoma cells. Brain Behav Immun. 2011 Mar;25(3):574- dependent transactivation of extracellular signal-regulated 83 kinase/cyclooxygenase-2 pathway. J Cell Biochem. 2008b Sep 1;105(1):53-60 Bravo-Calderón DM, Oliveira DT, Marana AN, Nonogaki S, Carvalho AL, Kowalski LP. Prognostic significance of beta- Rosenbaum DM, Rasmussen SG, Kobilka BK. The 2 adrenergic receptor in oral squamous cell carcinoma. structure and function of G-protein-coupled receptors. Cancer Biomark. 2011-2012;10(1):51-9 Nature. 2009 May 21;459(7245):356-63 Steenhuis P, Huntley RE, Gurenko Z, Yin L, Dale BA, Shang ZJ, Liu K, Liang de F. Expression of beta2- Fazel N, Isseroff RR. Adrenergic signaling in human oral adrenergic receptor in oral squamous cell carcinoma. J keratinocytes and wound repair. J Dent Res. 2011 Oral Pathol Med. 2009 Apr;38(4):371-6 Feb;90(2):186-92 Sivamani RK, Pullar CE, Manabat-Hidalgo CG, Rocke DM, Zhang P, He X, Tan J, Zhou X, Zou L. β-arrestin2 Carlsen RC, Greenhalgh DG, Isseroff RR. Stress-mediated mediates β-2 adrenergic receptor signaling inducing increases in systemic and local epinephrine impair skin prostate cancer cell progression. Oncol Rep. 2011 wound healing: potential new indication for beta blockers. Dec;26(6):1471-7 PLoS Med. 2009 Jan 13;6(1):e12 Loenneke JP, Wilson JM, Thiebaud RS, Abe T, Lowery Yang EV, Kim SJ, Donovan EL, Chen M, Gross AC, RP, Bemben MG. β2 Adrenoceptor signaling-induced Webster Marketon JI, Barsky SH, Glaser R. muscle hypertrophy from blood flow restriction: is there Norepinephrine upregulates VEGF, IL-8, and IL-6 evidence? Horm Metab Res. 2012 Jun;44(7):489-93 expression in human melanoma tumor cell lines: implications for stress-related enhancement of tumor This article should be referenced as such: progression. Brain Behav Immun. 2009 Feb;23(2):267-75 Oliveira DT, Bravo-Calderón DM. ADRB2 (adrenoceptor Bernabé DG, Tamae AC, Biasoli ÉR, Oliveira SH. Stress beta 2, surface). Atlas Genet Cytogenet Oncol Haematol. hormones increase cell proliferation and regulates 2014; 18(9):659-662. interleukin-6 secretion in human oral squamous cell

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Gene Section Review

IRF4 (interferon regulatory factor 4) Vipul Shukla, Runqing Lu Department of Genetics, Cell Biology & Anatomy, University of Nebraska Medical Center, Omaha, NE 68118, USA (VS, RL)

Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/IRF4ID231ch6p25.html DOI: 10.4267/2042/54034 This article is an update of : Rasi S, Gaidano G. IRF4 (interferon regulatory factor 4). Atlas Genet Cytogenet Oncol Haematol 2009;13(12):941-943.

mRNA is expressed at high levels in lymphoid Abstract tissues, in skin and in tonsils. Review on IRF4, with data on DNA/RNA, on the protein encoded and where the gene is implicated. Protein Identity Description Protein length: 451 amino acids. Other names: IRF-4, LSIRF, MUM1, NF-EM5 Calculated molecular weight of 51,8 kDa. HGNC (Hugo): IRF4 Expression Location: 6p25.3 IRF4 protein is expressed predominantly in blood Local order: IRF4 is located on chromosome 6 at cells. However, its expression can also be detected the telomeric extremity of the short arm, and lies in adipocytes and melanocytes. between the DUSP22 (dual specificity phosphatase In blood cells, expression of IRF4 can be detected 22) and EXOC2 (exocyst complex component 2) in T, B, DC and macrophages. Expression of IRF4 genes. in T and B cells is strongly induced by antigen Note receptor signaling. IRF4 belongs to the IRF (interferon regulatory Localisation factors) family of transcription factors and is a critical transcriptional regulator of immune system Nucleus. development and function. Function In the immune system, IRF4 is critical for DNA/RNA development and maturation of multiple lineages of blood cells. In T cells development, IRF4 is Description essential for the differentiation of Th1, Th2, Th9, Gene of 19,4 kb with 9 exons and 8 introns. Th17 and T reg subsets. In B lymphocytes, IRF4 Exon 1, the 5' part of exon 2 and the 3' part of exon promotes light chain rearrangement and 9 are non coding. transcription and is critical for B cell development Transcription at the pre-B stage. IRF4 antagonizes Notch signaling and limits the Length of the transcript is 5314 bp. size of marginal zone B cells (Simonetti et al., Coding sequence: CDS 114-1469. 2013).

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In addition, IRF4 is essential for class-switching course. IRF4 is obligatory required for the terminal and plasma cell differentiation. In B cells, IRF4 differetiation of mature B cells to plasma cells and interacts with Ets family of trancription factor has been shown to play a central role in the (PU.1/spi-B) through EICE site whereas in T cells, pathogenesis of MM. IRF4 is recurrently IRF4 interacts with AP-1 family of trancription translocated and juxtaposed to the IgH promoter factor (BATF) through AICE site. Also, IRF4 is t(6;14)(p25;q32) in a significant proportion (~21%) required for the differentiation of dendritic cells of MM cases. More commonly, IRF4 have been (DCs), particularly the CD11b(+) subset (Schlitzer shown to be overexpressed without genetic et al., 2013). In macrophages, IRF4 promotes the alterations in majority of MM cases and MM cells differentiation and polarisation to the M2 subtype are particularly sensitive to the down-regulation of also known as the tumor associated macrophages. IRF4. Recent studies have identified a role of IRF4 in Cytogenetics adipocyte biology. IRF4 has been shown to regulate t(6;14)(p25;q32) --> IRF4 - IgH. enzymes required for lipolysis in adipocytes. Therefore, an adipocyte specific deletion of IRF4 Hybrid/Mutated gene causes enhanced lipid synthesis, dysregulated lipid The translocation juxtaposes the IgH locus to the homeostasis eventually leading to obesity. IRF4 gene. Interestingly, in melanocytes IRF4 was recently Oncogenesis identified to cooperate with another transcription The precise mechanism for pathogenesis of MM in factor, MITF to positively regulate the expression presence of high levels of IRF4 is mediated by an of tyrosinase gene required for melanin synthesis. autoregulatory loop established between IRF4 and Additionally, the SNPs in the IRF4 gene locus have c-myc in MM cells. Recently, IRF4 has been shown been identified as risk alleles for developing to regulate caspase-10 leading to disruption of melanoma. normal autophagy mechanisms in MM cells Homology thereby, causing prolonged survival of these cells. Among IRF family members, IRF4 is highly Chronic lymphocytic leukemia (CLL) homologous to IRF8. Disease CLL is the most common adult leukemia in the Mutations western countries. It is a heterogeneous B-cell Germinal malignancy marked by progressive accumulation of CD5 positive mature B lymphocytes. A Genome SNPs in the IRF4 gene locus have been identified Wide Association Study (GWAS) recently in patients with chronic lymphocytic leukemia and identified SNPs in the 3' UTR of IRF4 gene locus in melanoma. patients with CLL. The individuals carrying the risk Somatic alleles harboring the SNPs have lower levels of Somatic mutations in DNA binding domain of IRF4 IRF4 and poorer outcomes compared to individuals have been identified in a small subset (1,5%) of carrying the non-risk allele. Another study chronic lymphocytic leukemia (CLL) patients. identified mutations in the DNA binding domain of IRF4 in a small subset (1,5%) of CLL cases. More Implicated in recently, using two distinct murine genetic models, it has been shown that low levels of IRF4 are Multiple myeloma (MM) causally related to the development of CLL. Disease Prognosis Multiple myeloma (MM) is a plasma cell derived CLL patients with low levels of IRF4 have malignancy with a particularly aggressive clinical aggressive disease course and poor prognosis.

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Cytogenetics Hodgkins lymphoma (HL) Although reciprocal translocations are extremely Disease rare in CLL, a translocation disrupting IRF4 gene Hodgkins lymphoma (HL) is an enigmatic B cell locus t(1;6)(p35.3;p25.2) was identified in a small malignancy that is characterized by lack of subset of CLL patients with aggressive disease. expression of several B cell markers. Hybrid/Mutated gene The Hodgkin and Reed Sternberg (HRS) cells Mutations in the DNA binding domain of IRF4 present in HL cases are presumably derived from with a yet undefined function in B cells were germinal center B cells. IRF4 is overexpressed in identified in a small subset of CLL cases. majority of classical HL cases and is shown to Oncogenesis mediate the survival of these cells. Paradoxically, The precise mechanism for oncogenesis of CLL in the SNPs in IRF4 linked to its lower expression presence of low levels of IRF4 is not yet known. levels and associated with the development of CLL are also shown to be linked to the risk of Diffused large B cell lymphoma developing HL. (DLBCL) Oncogenesis Disease Whether the overexpression of IRF4 in HRS cells Diffuse large B cell lymphoma represents a of HL is causal is unclear. However, some studies heterogeneous malignancy that arises have linked the survival and proliferation of HRS spontaneously or develop from pre-existing cells to the expression of IRF4. leukemia. On the basis of gene expression profiling DLBCL is divided into three distinct subtypes Primary cutaneous anaplastic large namely the germinal center subtype (GCB), the cell lymphoma (C-ALCL) activated B cell subtype (ABC) and the mediastinal Disease subtype. The three subtypes presumably arise from Primary cutaneous anaplastic large cell lymphoma three distinct B cell subtypes. IRF4 is primarily (C-ALCL) is a T cell lymphoma with an indolent overexpressed in the ABC type of DLBCL while disease course and presence of tumor lesions in the GCB subtype is marked by lower expression of skin. The lesions in C-ALCL almost never spread IRF4. extra-cutaneously and often regress spontaneously. Prognosis IRF4 is overexpressed in C-ALCL but not in the IRF4 is overexpressed in the ABC type DLBCL more aggressive form of the disease known as which is most aggressive form of DLBCL and have peripheral T cell lymphoma not otherwise specified poorer patient outcomes compared to other (PTCL-NOS). The overexpression of IRF4 in some subtypes. cases is associated with a recurrent translocations a subset of them placing the IRF4 gene next to the T Cytogenetics cell receptor alpha (TCRA) promoter IRF4 is overexpressed in a small group of patients t(6;14)(p25;q11.2). Other translocations identified with a reciprocal translocation between IgG locus do not involve TCRA. and the IRF4 t(1;6)(p35.3;p25.2). The patients carrying the translocation primarily belong to GCB Cytogenetics or follicular lymphoma grade 3 type is associated IRF4 is translocated primarily in the C-ALCL with favorable patient outcomes. however the precise breakpoints are not defined. In a small subset of the cases with translocations IRF4 Oncogenesis is juxtaposed to the TCRA locus IRF4 induces the expression of transcription factor t(6;14)(p25;q11.2). Blimp-1 and directly suppresses the expression of Bcl-6 to allow terminal differentiation of activated B cell acute lymphoblastic leukemia B cells to plasma cells. However, this molecular (B-ALL) network is short circuited in ABC DLBCL by recurrent mutational inactivation of Blimp-1. Disease Additionally, mutations located in the promoter B cell acute lymphoblastic leukemia (B-ALL) is a region of Bcl-6 that disrupt the IRF4 binding sites B cell malignancy derived from early B cells. IRF4 and leads to enhanced expression of Bcl-6 were is shown to play a tumor suppressive role in B- identified in a small group of patients. These ALL. IRF4 is shown to suppress the oncogenesis of genetic events disrupt the molecular network both BCR-ABL and c-myc induced B-ALL. required for plasma cell differentiation. However, Oncogenesis the precise functional role of IRF4 in pathogenesis IRF4 inhibits B-ALL by regulating the expression of ABC type DLBCL is not well defined. of negative regulators of cell cycle p27.

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Chronic myeloid leukemia (CML) the screen map to a putative enhancer region in the IRF4 gene locus. Disease Chronic myeloid leukemia (CML) is a Oncogenesis myeloproliferative disorder marked by clonal Recently, the SNP identified in IRF4 locus were expansion of granulocytes. It is associated with a demonstrated to decrease IRF4 expression by hallmark translocation and presence of a fusion disruption of specific transcription factor binding BCR-ABL protein in majority of patients. IRF4 is sites. Additionally, IRF4 corroborates with shown to be underexpressed in CML patients along micropthalmia associated transcription factor with its highly homologous family member IRF8. (MITF) to regulate the expression of enzyme However the functional role of IRF4 in CML is not tyrosinase responsible for melanin production. well characterized. These studies point towards a critical role for IRF4 in melanocyte biology and also its association with Virus implicated malignancies skin cancer. Disease Viruses like Epstein Barr virus (EBV), human T References cell leukemia virus-1 (HTLV1) and Kaposi Grossman A, Mittrücker HW, Nicholl J, Suzuki A, Chung S, Sarcoma associated herpes virus (KSHV/HHV-8) Antonio L, Suggs S, Sutherland GR, Siderovski DP, Mak are implicated in B cell malignancies, adult T cell TW. Cloning of human lymphocyte-specific interferon leukemia (ATL) and primary effusion lymphoma regulatory factor (hLSIRF/hIRF4) and mapping of the gene (PEL) respectively. The proteins encoded by these to 6p23-p25. Genomics. 1996 Oct 15;37(2):229-33 viruses, directly or indirectly activate NF-kB Iida S, Rao PH, Butler M, Corradini P, Boccadoro M, Klein signaling which in turn activates the expression of B, Chaganti RS, Dalla-Favera R. Deregulation of IRF4. As a result IRF4 is overexpressed in these MUM1/IRF4 by chromosomal translocation in multiple myeloma. Nat Genet. 1997 Oct;17(2):226-30 virus implicated malignancies. The knockdown of IRF4 in EBV transformed B cells lead to down- Mittrücker HW, Matsuyama T, Grossman A, Kündig TM, Potter J, Shahinian A, Wakeham A, Patterson B, Ohashi regulation of genes involved in cellular PS, Mak TW. Requirement for the transcription factor proliferation. The role of IRF4 in HTLV-1 induced LSIRF/IRF4 for mature B and T lymphocyte function. ATL is not clear however few reports indicate its Science. 1997 Jan 24;275(5299):540-3 involvement in regulation of cell cycle associated Tsuboi K, Iida S, Inagaki H, Kato M, Hayami Y, Hanamura genes. The role of IRF4 in KSHV induced kaposi's I, Miura K, Harada S, Kikuchi M, Komatsu H, Banno S, sarcoma and PEL is ambiguous. KSHV encodes Wakita A, Nakamura S, Eimoto T, Ueda R. MUM1/IRF4 viral homologs of cellular IRFs called vIRFs. The expression as a frequent event in mature lymphoid malignancies. Leukemia. 2000 Mar;14(3):449-56 vIRF4 is shown to inhibit the function of cellular IRF4 leading to induction of lytic cycle for KSHV Chang CC, Lorek J, Sabath DE, Li Y, Chitambar CR, Logan B, Kampalath B, Cleveland RP. Expression of replication. MUM1/IRF4 correlates with clinical outcome in patients Oncogenesis with B-cell chronic lymphocytic leukemia. Blood. 2002 Dec The role of IRF4 in these viral implicated 15;100(13):4671-5 malignancies is still unclear. However, the Falini B, Mason DY. Proteins encoded by genes involved activation status of NF-kB by these viruses in chromosomal alterations in lymphoma and leukemia: clinical value of their detection by immunocytochemistry. invariably co-relates with IRF4 expression in these Blood. 2002 Jan 15;99(2):409-26 cells. Ito M, Iida S, Inagaki H, Tsuboi K, Komatsu H, Yamaguchi Skin cancer M, Nakamura N, Suzuki R, Seto M, Nakamura S, Morishima Y, Ueda R. MUM1/IRF4 expression is an Disease unfavorable prognostic factor in B-cell chronic lymphocytic Skin cancer is associated with malignant or non- leukemia (CLL)/small lymphocytic lymphoma (SLL). Jpn J malignant lesions on the skin. Based on the cell of Cancer Res. 2002 Jun;93(6):685-94 origin, skin cancer can be divided into three types: Mamane Y, Grandvaux N, Hernandez E, Sharma S, basal cell carcinoma, squamous cell carcinoma and Innocente SA, Lee JM, Azimi N, Lin R, Hiscott J. melanoma. The differential skin pigmentation Repression of IRF-4 target genes in human T cell leukemia induced by melanin production alters the risk for virus-1 infection. Oncogene. 2002 Oct 3;21(44):6751-65 skin cancer. Particularly individuals with light skin Nishiya N, Yamamoto K, Imaizumi Y, Kohno T, tones and hence low melanin secretion are more Matsuyama T. Identification of a novel GC-rich binding protein that binds to an indispensable element for predisposed to developing skin cancer. Until constitutive IRF-4 promoter activity in B cells. Mol recently there were no known reports for a role of Immunol. 2004 Jul;41(9):855-61 IRF4 in melanocytes. However recently, SNPs Honma K, Udono H, Kohno T, Yamamoto K, Ogawa A, identified in the IRF4 gene locus have been shown Takemori T, Kumatori A, Suzuki S, Matsuyama T, Yui K. to be associated with skin pigmentation and the risk Interferon regulatory factor 4 negatively regulates the for developing skin cancer. The SNP identified in production of proinflammatory cytokines by macrophages

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Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 667 Atlas of Genetics and Cytogenetics in Oncology and Haematology

OPEN ACCESS JOURNAL INIST -CNRS

Gene Section Review

PLCG1 (Phospholipase C, Gamma 1) Rebeca Manso Pathology Department, Fundacion Conchita Rabago, IIS "Fundacion Jimenez Diaz", E-28040 Madrid, Spain (RM)

Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/PLCG1ID44163ch20q12.html DOI: 10.4267/2042/54035 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract Expression PLCG1 is expressed ubiquitously, especially in the Review on PLCG1, with data on DNA/RNA, on the brain, thymus, intestine and lungs. Additionally, protein encoded and where the gene is implicated. PLCG1 is overexpressed in numerous cancer types such as human colorectal cancer (Noh et al., 1994), Identity breast carcinoma (Arteaga et al., 1991), prostate Other names: NCKAP3, PLC-II, PLC1, PLC148, carcinoma (Peak et al., 2008), familial adenomatous PLCgamma1 polyposis (Park et al., 1994) and human skins under hypeproliferative conditions (Nanney et al., 1992). HGNC (Hugo): PLCG1 Location: 20q12 Localisation Local order: From the cytosol. PLCG1 localizes predominantly in the plasmatic membrane, cytoplasm and nucleus. DNA/RNA Function Description PLCG1 is a protein involved in multiple cellular The PLCG1 gene spans 38.762 kb on the genomic processes. A potent inhibitor of PLCG1 (U-73122) DNA. The gene includes 32 exons. has been reported to inhibit PLCG1-dependent processes in cells (Smith et al., 1990; Thompson et Transcription al., 1991; Thomas et al., 2003; Li et al., 2005). There are two transcript variants: 5205 bp (isoform The inhibition of PLCG1 may be an important a) and 5202 bp (isoform b). mechanism for an antiproliferative effect on the human cancer cells. Protein Role in the production of the second messenger molecules: PLCG1 mediates the production of Description diacylglycerol (DAG) and inositol 1,4,5- The PLCG1 protein encodes two alternative trisphosphate (IP3) from the hydrolysis of isoforms: variant a (P19174-1)-1290 amino acids, phosphatidynositol-4,5-bisphosphate (PIP 2) 148.53 Da; variant b (P19174-2)-1291 amino acids, (Williams et al., 1996). These second messengers 148.66 Da. are essential for T cell activation (Lin et al., 2001).

Figure 1. Schematic diagram of PLCG1 location on chromosome 20. PLCG1 localizes to chromosome 20q12, which is represented graphically. PLCG1 gene spans 38.762 kb on the genomic DNA.

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Figure 2. Schematic representation of the domains of PLCG1. The protein contains eight domains, four of which are unique to PLCG family. The PLCG 'specific array' of domains, comprising a "split" PH domain flanking two tandem SH2 domains and one SH3 domain, is inserted between the two halves (X and Y) of the TIM-barrel catalytic domain. Several other domains including two PH domains, one C2 domain and one EF hand motifs. The numering of the amino acid residues is for human PLCG1 (Suh et al., 2008; Bunney and Katan, 2011).

Role in cellular proliferation: PLCG1 is associated dependent on PLCG1 activation (Wozniak et al., with tumor development, and it is overexpressed in 2006). some tumors (Shin et al., 2007). This Role in transformation: PLCG1 interacts with overexpression stimulates MMP-3 expression. Middle tumor antigen (MT). PLCG1 is required for metastasis development The tyrosine phosphorylation level of PLCG1 is (Sala et al., 2008). elevated in cells expressing wild type MT but not in Role in angiogenesis: PLCG1 plays an important cells expressing Tyr322 →Phe MT (Su et al., 1995). role in angiogenesis (Husain et al., 2010). PLCG1 Role in autoimmune symptoms: PLCG1 deficiency is activated by vascular endothelial growth factor impairs the development and function regulatory receptor-2 (VEGFR-2) in endothelial cells (Singh et cells (FoxP3+), causing inflammatory/autoimmune al., 2007) and in neoplastic Barrett's cells (Zhang et symptoms (Fu et al., 2010). al., 2013). Role in the regulation of intracellular signaling: Homology PLCG1 plays a role in mediating T-cell activities The protein contains eight domains, four of which downstream of TCR activity. are unique to PLCG family (Suh et al., 2008). PLCG1 can be activated by receptor tyrosine The PLCG 'specific array' of domains, comprising a kinases: EGFR (Nishibe et al., 1990; Wu et al., "split" PH domain flanking two tandem SH2 2009), PDGFR (Larose et al., 1993), FGFR (Peters domains and one SH3 domain, is inserted between et al., 1992), NGFR (Middlemas et al., 1994) and the two halves (X and Y) of the TIM-barrel HGFR (Davies et al., 2008). PLCG1 is a molecule catalytic domain (Bunney and Katan, 2011). associate with lipid rafts, it translocates from the Several other domains including two PH domains, cytosol to lipid rafts during TCR signaling (Verí et one C2 domain and one EF hand motifs (Suh et al., al., 2001). 2008). Role in the mobilization of Ca 2+ : this process is to activate phosphatase calcineurin, which in turn Mutations dephosphorylates and activates NFAT (Rao et al., 1997). Somatic Truncation of the N terminus of Vav1 is 99 mutations have been described in the PLCG1 accompanied by a decrease in PLCG1 gene, according to the Catalogue of Somatic phosphorylation and this inhibits calcium Mutations in Cancer (COSMIC) database. mobilization (Knyazhitsky et al., 2012). De novo mutation has been described in patients Role in cytoskeleton: PLCG1 plays a role in actin with Cutaneous T-cell lymphoma (CTCL): S345F reorganization (Pei et al., 1996; Wells, 2000; Wang (10/53 analyzed CTCL samples, 19%) (Vaqué et et al., 2007; Li et al., 2009). al., 2014). Role in adhesion and migration: PLCG1 mediates cell adhesion and migration through an undefined mechanism (Wang et al., 2007; Crooke et al., Implicated in 2009). PLCG1 plays a role in integrin-mediated cell motility processes (Jones et al., 2005). Breast cancer Role in apoptosis: PLCG1 is proteolytically cleaved Oncogenesis by group II caspases especially by caspase-3 and Overexpression of PLCG1 is a marker of caspase-7 during apoptosis. This results in the loss development of metastases in breast cancer of receptor-mediated tyrosine phosphorylation (Bae (Lattanzio et al., 2013). et al., 2000). PLCG1 plays a protective role in Loss of PLCG1 in part mimicked the effect of miR- H2O2-induced PC12 cells death (Yuan et al., 2009). 200b/miR-c/miR-429 overexpression in viability, The Fas-mediated apoptosis requires endoplasmic apoptosis and EGF-driven cell invasion of breast reticulum-mediated calcium release in a mechanism cancer cells (Uhlmann et al., 2010).

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Colorectal cancer signaling towards NFAT activation (Vaqué et al., 2014). Oncogenesis PLCG1 has a potencial role in colon cancer Brain disorders (Nomoto et al., 1995; Li et al., 2005; Reid et al., Note 2009). The activity of PLCG1 is reduced in STAT3 Jang et al., 2013. Y705F mutant colorectal cancer cells (Zhang et al., Oncogenesis 2011), it shows that there is crosstalk between STAT3 and PLCG1 signaling pathways. PLCG1 is highly expressed in brain. Abnormal expression and activation of PLCG1 appears in Prostate carcinoma epilepsy (He et al., 2010), bipolar disorder (Løvlie Oncogenesis et al., 2001), depression (Dwivedi et al., 2005), PLCG1 has a role in the regulation of PC3LN3 Huntington's disease (Giralt et al., 2009) and (human prostate carcinoma cells) cell adhesion that Alzheimer's disease (Shimohama et al., 1995). appears to be independent of its effects on tumour Myocardial dysfunction in sepsis cell chemotactic migration and spreading in Oncogenesis response to extracellular matrix (Peak et al., 2008). PLCG1 signaling induces cardiac TNF-alpha Gastric cancer expression and myocardial dysfunction during Oncogenesis Lipopolysaccharide (LPS) stimulation. Inhibition of PLCG1 plays a role in RhoGDI2-mediated cisplatin PLCG1 decreased cardiac TNF-alpha expression resistance and cell invasion in gastric cancer (Cho and LPS-induced myocardial dysfunction was also et al., 2011). attenuated (Peng et al., 2008). Squamous cell carcinoma (SCC) To be noted Oncogenesis PLCG1 is a downstream target of EGFR signaling. Note PLCG1 is required for EGFR-induced SCC cell miR that target PLCG1: PLCG1 is target of mitogenesis (Xie et al., 2010). different microRNAs, according to the bioinformatic algorithms microRNA Oral potentially malignant lesions (microRNA.org). (OPLs) Oncogenesis References PLCG1 is highly expressed in oral cancer lesions Nishibe S, Wahl MI, Wedegaertner PB, Kim JW, Rhee SG, compared with normal oral mucosa (Ma et al., Carpenter G. Selectivity of phospholipase C 2013). phosphorylation by the epidermal growth factor receptor, the insulin receptor, and their cytoplasmic domains. Proc Esophageal adenocarcinoma Natl Acad Sci U S A. 1990 Jan;87(1):424-8 Oncogenesis Smith RJ, Sam LM, Justen JM, Bundy GL, Bala GA, PLCG1-PKC-ERK pathway promotes proliferation Bleasdale JE. Receptor-coupled signal transduction in human polymorphonuclear neutrophils: effects of a novel and it is activated by VEGFR-2 in neoplastic inhibitor of phospholipase C-dependent processes on cell Barrett's cells (Zhang et al., 2013). responsiveness. J Pharmacol Exp Ther. 1990 Pheochromocytoma (PC) May;253(2):688-97 Arteaga CL, Johnson MD, Todderud G, Coffey RJ, Oncogenesis Carpenter G, Page DL. Elevated content of the tyrosine PLCG1 plays a role in apoptosis of PC12 cells kinase substrate phospholipase C-gamma 1 in primary induced by H 2O2 (Yuan et al., 2009). human breast carcinomas. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10435-9 Glioblastoma Thompson AK, Mostafapour SP, Denlinger LC, Bleasdale Oncogenesis JE, Fisher SK. The aminosteroid U-73122 inhibits PLCG1 is associated with lifetime and overall muscarinic receptor sequestration and phosphoinositide hydrolysis in SK-N-SH neuroblastoma cells. A role for Gp survival in glioblastoma and it can be a novel in receptor compartmentation. J Biol Chem. 1991 Dec biomarker of this desease (Serão et al., 2011). 15;266(35):23856-62 Cutaneous T-cell lymphoma (CTCL) Nanney LB, Gates RE, Todderud G, King LE Jr, Carpenter G. Altered distribution of phospholipase C-gamma 1 in Oncogenesis benign hyperproliferative epidermal diseases. Cell Growth The mutation in the catalytic domain of PLCG1 Differ. 1992 Apr;3(4):233-9 (S345F) is detection in patients with CTCL. PLCG1 Peters KG, Marie J, Wilson E, Ives HE, Escobedo J, Del mutants induced enhanced PLCG1 downstream Rosario M, Mirda D, Williams LT. Point mutation of an FGF

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J Neurosci. 2010 Jan;42(1):47-52 May 5;30(18):6188-96 Vaqué JP, Gómez-López G, Monsálvez V, Varela I, Husain D, Meyer RD, Mehta M, Pfeifer WM, Chou E, Martínez N, Pérez C, Domínguez O, Graña O, Rodríguez- Navruzbekov G, Ahmed E, Rahimi N. Role of c-Cbl- Peralto JL, Rodríguez-Pinilla SM, González-Vela C, Rubio- dependent regulation of phospholipase Cgamma1 Camarillo M, Martín-Sánchez E, Pisano DG, Papadavid E, activation in experimental choroidal neovascularization. Papadaki T, Requena L, García-Marco JA, Méndez M, Invest Ophthalmol Vis Sci. 2010 Dec;51(12):6803-9 Provencio M, Hospital M, Suárez-Massa D, Postigo C, San Segundo D, López-Hoyos M, Ortiz-Romero PL, Piris MA, Uhlmann S, Zhang JD, Schwäger A, Mannsperger H, Sánchez-Beato M. PLCG1 mutations in cutaneous T-cell Riazalhosseini Y, Burmester S, Ward A, Korf U, Wiemann lymphomas. Blood. 2014 Mar 27;123(13):2034-43 S, Sahin O. miR-200bc/429 cluster targets PLCgamma1 and differentially regulates proliferation and EGF-driven Zhang Q, Yu C, Peng S, Xu H, Wright E, Zhang X, Huo X, invasion than miR-200a/141 in breast cancer. Oncogene. Cheng E, Pham TH, Asanuma K, Hatanpaa KJ, Rezai D, 2010 Jul 29;29(30):4297-306 Wang DH, Sarode V, Melton S, Genta RM, Spechler SJ, Souza RF. Autocrine VEGF signaling promotes Xie Z, Chen Y, Liao EY, Jiang Y, Liu FY, Pennypacker SD. proliferation of neoplastic Barrett's epithelial cells through a Phospholipase C-gamma1 is required for the epidermal PLC-dependent pathway. Gastroenterology. 2014 growth factor receptor-induced squamous cell carcinoma Feb;146(2):461-72.e6 cell mitogenesis. Biochem Biophys Res Commun. 2010 Jun 25;397(2):296-300 This article should be referenced as such: Bunney TD, Katan M. PLC regulation: emerging pictures Manso R. PLCG1 (Phospholipase C, Gamma 1). Atlas for molecular mechanisms. Trends Biochem Sci. 2011 Genet Cytogenet Oncol Haematol. 2014; 18(9):668-672.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 672 Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Gene Section Review

SLC1A5 (solute carrier family 1 (neutral amino acid transporter), member 5) Cesare Indiveri, Lorena Pochini, Michele Galluccio, Mariafrancesca Scalise Department DiBEST (Biologia, Ecologia, Scienze della Terra) Unit of Biochemistry and Molecular Biotechnology, University of Calabria, 87036 Arcavacata di Rende, Italy (CI, LP, MG, MS)

Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/SLC1A5ID42313ch19q13.html DOI: 10.4267/2042/54036 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

constituted by 1737 nucleotides and differs in the 5' Abstract UTR from the variant NM_005628. Review on human SLC1A5, with data on In NM_001145144 the translation starts DNA/RNA, on the protein encoded and downstream the third exon generating a shorter pathological and physiological implications. peptide of 313 aa. The third isoform NM_ 001145145 has 1927 Identity nucleotides and lacks the first exon. It presents a different translation start at 5', coding a peptide of Other names: AAAT, ASCT2, ATBO, M7V1, 339 amino acids. A longer transcript, M7VS1, R16, RDRC XM_005259167, is reported only in NCBI HGNC (Hugo): SLC1A5 database. Location: 19q13.32 It has been identified by automated computational Local order: Orientation: minus strand. analysis. More than 400 SNP(s), both in coding and non-coding regions of the SLC1A5 gene, are DNA/RNA reported in dbSNP database (dbSNP). More than 40 are responsible of amino acid substitutions with Description unknown significance. Only the variant SLC1A5- The SLC1A5 gene, located at 19q13.3, counts P17A (rs3027956) is associated with breast cancer 28692 nucleotides with 8 exons. It has been found (Savas et al., 2006). A region constituted by 907 bp in 56 different organisms (NCBI). The gene upstream of the ASCT2 gene possesses promoter encodes a protein involved in sodium-dependent activity (Bungard and McGivan, 2004). In this neutral amino acid transport (Kekuda et al., 1996; region the following putative elements have been Pingitore et al., 2013). identified: an amino acid-regulatory element, a consensus site for binding of the transcription factor Transcription activator protein 1 (AP1) and a consensus binding Three isoforms (transcripts) are reported either on sites for nuclear and hepatocyte nuclear factors. NCBI and Ensembl databases for SLC1A5 human gene, deriving from different translation start. They Pseudogene differ in length, particularly at 5' extremity. The The gene is virtually present in all vertebrates. The first variant NM_005628 represents the longest better known orthologous of the human gene are transcript, constituted by 2873 nucleotides and 8 those from rat, mouse and rabbit. Identity between exons. This transcript encodes a peptide of 541 the human and rat, mouse, rabbit are 79%, 82% and amino acids. The second variant NM_001145144 is 85%, respectively.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 673 SLC1A5 (solute carrier family 1 (neutral amino acid transporter), member 5) Indiveri C, et al.

Figure 1. Isoforms of SLC1A5 gene. The three isoforms are present in the minus strand of the chromosome 19 in position 19q13.3. NM_005628: isoform one, encodes for the longest peptide and is constituted by 8 exons; NM_001145144: isoform two, due to alternative splicing is characterized by only four exons; NM_ 001145145: isoform three presents seven exons. The nucleotide sequence is depicted as black lines. Coding nucleotides and untranslated (UTR) regions are indicated by red and white boxes, respectively. Exons are indicated by roman numbers.

Protein Function Transport mediated by the human ASCT2 has been Description originally studied in intact cell systems over- 541 amino acids; molecular mass 56598,34 Da. expressing the transport protein (Kekuda et al., Human SLC1A5 is a permease (membrane 1996; Kekuda et al., 1997). transporter). Recently, hASCT2 was over-expressed in the yeast The 3D structure is not available. Homology P. pastoris, purified and reconstituted in artificial modeling highlights a structure similar to that of the phospholipid vesicles (proteoliposomes), in absence glutamate transporter of P. horikoshii (1XFH). N- of other interfering transporters. and C-terminal ends are intracellular. Potential site All experimental systems concur in demonstrating of N-glycosylation and phosphorilation are that hASCT2 is an obligate exchanger of neutral predicted. amino acid. In the structural model, at least one glycosylation This antiport requires the presence of extracellular site is extracellular and the phosphorilation sites are Na + which cannot be substituted by Li + or K +. The + intracellular (Fig. 2). Na ex :amino acid ex stoichiometry of the human Expression transporter is likely to be 1:1. Competition studies on 3H-glutamine, 3H-threonine or 3H-alanine Human SLC1A5 has been originally named ASCT2 transport performed in cells indicated that other from AlaSerCysTransporter2 or ATB0. potential substrates of hASCT2 are valine, leucine, The acronym ASCT2 is the most frequently used to serine, cysteine, asparagine, methionine, isoleucine, designate this transport system. tryptophan, histidine, phenylalanine. While It is expressed in many tissues, including brain, glutamate, lysine, arginine along with MeAIB [ α- (Bröer and Brookes, 2001; Deitmer et al., 2003; (methylamino)isobutyric acid] and BCH [2- Gliddon et al., 2009). aminobicyclo-(2,2,1)-heptane-2-carboxylic acid] There is functional evidence of the expression of are neither transported nor inhibit hASCT2. ASCT2 in kidney and intestine (Bode, 2001). Experiments with radioactive compounds Besides Caco-2 cells, apparently, also the HT-29 confirmed the competition data (Torres-Zamorano intestinal cell line functionally expresses ASCT2 et al., 1998). In proteoliposomes, inhibition has (Kekuda et al., 1996; Kekuda et al., 1997). been confirmed for most but not for all of the amino Poly(A)1 RNA isolated from several tissues of acids. Moreover, proteoliposome studies human origin revealed expression in placenta, lung, highlighted an asymmetric specificity for amino skeletal muscle, kidney, and pancreas (Kekuda et acids allowing to distinguish the amino acids al., 1996). inwardly transported (alanine, cysteine, valine, Localisation methionine) from those bi-directionally transported The protein is localized in the plasma membrane. (glutamine, serine, asparagine, and threonine).

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 674 SLC1A5 (solute carrier family 1 (neutral amino acid transporter), member 5) Indiveri C, et al.

Figure 2. Homology structural model of hASCT2. Ribbon diagram viewing of the transporter from the lateral side. The model was built using the glutamate transporter Glpth from Pyrococcus horikoshii crystal structure (1XFH) as the template by Modeller V9.13. The homology model was represented using SpdbViewer 4.01. Asn 163 and 212, predicted as glycosilation sites, are highlighted in blue; Ser 183, 261 and Thr 206, 207, 329, predicted as phosphorilation sites are highlighted in red and orange, respectively. Prediction according to Scan Prosite.

The functional asymmetry was also confirmed by regulation of mTOR pathway, translation and the kinetic analysis of [ 3H]glutamine/glutamine autophagy. The transporter regulates an increase in antiport: different Km values were measured on the the intracellular concentration of glutamine which external and internal sides of proteoliposomes, is then used by another plasma membrane 0,097 and 1,8 mM, respectively. transporter, named LAT1 (SLC7A5) (Galluccio et The SH reagents HgCl 2, mersalyl and pOHMB al., 2013) as efflux substrate to regulate the uptake potently inhibited hASCT2 mediated transport of extracellular leucine with subsequent activation (Pingitore et al., 2013). of mTORC1 (Nicklin et al., 2009). Moreover, it has The physiological role of hASCT2 consists in been proposed that a group of retroviruses providing cells with some neutral amino acids specifically uses the hASCT2 as a common cell exporting others on the basis of the metabolic need surface receptor following a co-evolution of cells consistently with the intra and extracellular phenomenon. The orthologous murine transporter amino acid concentrations. In brain, particularly, mASCT2 is inactive as a viral receptor (Marin et hASCT2 contributes to glutamine homeostasis of al., 2003). neurons and astrocytes. On the basis of experiments performed with animal models, it was hypothesized Implicated in that hASCT2 mediates efflux of glutamine from astrocytes, a process that is critical for the Molecular basis of cancerogenesis functioning of the glutamate-glutamine cycle to Note recover synaptically released glutamate in exchange Tumor cells acquire altered metabolism. Due to with glutamine efflux (Bröer et al., 1999). The these changes, the expression of membrane glutamine-glutamate cycle has been shown also in transporters involved in providing nutrients is placenta. Glutamine crosses the placenta and enters altered. The plasma membrane transporter for the fetal liver where it is deamidated to glutamate. glutamine ASCT2 has been clearly associated to About 90% of glutamate generated by the liver is cancer development and progression, together with taken up by the placenta and used in the another amino acid membrane transporter, LAT1 metabolism. The glutamine-glutamate cycle specific for glutamine and other neutral amino acids between the placenta and the fetal liver is (Fuchs and Bode, 2005). The energetic needs of obligatory for the generation of NADPH in the cancer cells are different from normal ones due to placenta (Torres-Zamorano et al., 1998). Among the Warburg effect. According to this phenomenon other functions reported for hASCT2 there is the ATP derives from anaerobic glycolisis bypassing

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 675 SLC1A5 (solute carrier family 1 (neutral amino acid transporter), member 5) Indiveri C, et al.

mitochondrial function (Ganapathy et al., 2009). In Breast cancer this scenario glutamine provided by means of Note ASCT2 and LAT1 transport function sustains tumor In breast cancer ASCT2 has been found over growth and signaling through mTOR pathway expressed together with other proteins related to (Nicklin et al., 2009). glutamine metabolism like glutamminase and The importance of ASCT2 in this network is glutamate dehydrogenase (Kim et al., 2012). revealed by induction of apoptosis when silencing The study revealed that this metabolism is essential its gene in human hepatoma cells (Fuchs et al., for sustaining breast cancer development and that 2004). the protein levels are different according to In the following paragraphs specific examples of different subtypes of cancer. The subtype HER2 human cancers are reported. showed the highest level of glutamine related Prostate cancer proteins and that the basal-like breast cancers are Note more dependent on glutamine compared to luminal- Tissue microarray technology (TMA) has been used likeones. for studying ASCT2 in normal prostatic tissue, in Other diseases benign prostatic hyperplasia and in prostate Note adenocarcinoma. Due to importance of glutamine in cell metabolism In particular, a negative prognosis and a shorter and the chromosomal localization of SLC1A5 gene, time of recurrence for adenocarcinoma were several association studies have been conducted to associated to hASCT2 expression. Moreover, a ascertain the involvement of hASCT2 in more aggressive behavior of adenocarcinoma is pathologies like cystinuria, cystic fibrosis, described (Li et al., 2003). schizophrenia, Hartnup disorder and pre-eclampsia. Colorectal carcinoma However, no genetic associations have been Note revealed. The expression of ASCT2 in colorectal carcinoma is normally associated to a decrease of percentage To be noted in patient survival (Witte et al., 2002). Note Neuroblastoma and glioma Aknowledgements: This work was supported by Note funds from: Programma Operativo Nazionale [PON Neuroblastoma are childhood tumors very often 01_00937] "Modelli sperimentali Biotecnologici benign. In some cases, however, neuroblastoma integrati per lo sviluppo e la selezione di molecole became malignant. One of the biological marker of di interesse per la salute dell'uomo", Ministero this second category is the increased uptake of Istruzione Università e Ricerca (MIUR). glutamine and other neutral aminoacids via ASCT2 (Wasa et al., 2002). Human glioma C6 cells have References been demonstrated to mediate uptake of glutamine Kekuda R, Prasad PD, Fei YJ, Torres-Zamorano V, Sinha via ASCT2 (Dolinska et al., 2003). S, Yang-Feng TL, Leibach FH, Ganapathy V. Cloning of the sodium-dependent, broad-scope, neutral amino acid Hepatoma transporter Bo from a human placental choriocarcinoma Note cell line. J Biol Chem. 1996 Aug 2;271(31):18657-61 Hepatocell carcinoma (HCC) is the most common Kekuda R, Torres-Zamorano V, Fei YJ, Prasad PD, Li HW, malignant tumor of liver and one of the main cause Mader LD, Leibach FH, Ganapathy V. Molecular and functional characterization of intestinal Na(+)-dependent of death. A study reported that higher rate of neutral amino acid transporter B0. Am J Physiol. 1997 glutamine uptake via ASCT2 is a common feature Jun;272(6 Pt 1):G1463-72 of six examined hepatoma cell line (Bode et al., Torres-Zamorano V, Leibach FH, Ganapathy V. Sodium- 2002; Fuchs et al., 2004). dependent homo- and hetero-exchange of neutral amino acids mediated by the amino acid transporter ATB degree. Lung cancer Biochem Biophys Res Commun. 1998 Apr 28;245(3):824-9 Note Bröer A, Brookes N, Ganapathy V, Dimmer KS, Wagner ASCT2 has been found over expressed in lung CA, Lang F, Bröer S. The astroglial ASCT2 amino acid cancer by proteomic approach and then confirmed transporter as a mediator of glutamine efflux. J Neurochem. 1999 Nov;73(5):2184-94 at molecular level. Pharmacologic and genetic targeting of ASCT2 decreased cell growth and Bröer A, Wagner C, Lang F, Bröer S. Neutral amino acid transporter ASCT2 displays substrate-induced Na+ viability in lung cancer cells, an effect mediated in exchange and a substrate-gated anion conductance. part by mTOR signaling (Hassanein et al., 2013). Biochem J. 2000 Mar 15;346 Pt 3:705-10

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 676 SLC1A5 (solute carrier family 1 (neutral amino acid transporter), member 5) Indiveri C, et al.

Bode BP. Recent molecular advances in mammalian Fuchs BC, Bode BP. Amino acid transporters ASCT2 and glutamine transport. J Nutr. 2001 Sep;131(9 Suppl):2475S- LAT1 in cancer: partners in crime? Semin Cancer Biol. 85S; discussion 2486S-7S 2005 Aug;15(4):254-66 Bröer S, Brookes N. Transfer of glutamine between Savas S, Schmidt S, Jarjanazi H, Ozcelik H. Functional astrocytes and neurons. J Neurochem. 2001 nsSNPs from carcinogenesis-related genes expressed in May;77(3):705-19 breast tissue: potential breast cancer risk alleles and their distribution across human populations. Hum Genomics. Bode BP, Fuchs BC, Hurley BP, Conroy JL, Suetterlin JE, 2006 Mar;2(5):287-96 Tanabe KK, Rhoads DB, Abcouwer SF, Souba WW. Molecular and functional analysis of glutamine uptake in Ganapathy V, Thangaraju M, Prasad PD. Nutrient human hepatoma and liver-derived cells. Am J Physiol transporters in cancer: relevance to Warburg hypothesis Gastrointest Liver Physiol. 2002 Nov;283(5):G1062-73 and beyond. Pharmacol Ther. 2009 Jan;121(1):29-40 Wasa M, Wang HS, Okada A. Characterization of L- Gliddon CM, Shao Z, LeMaistre JL, Anderson CM. Cellular glutamine transport by a human neuroblastoma cell line. distribution of the neutral amino acid transporter subtype Am J Physiol Cell Physiol. 2002 Jun;282(6):C1246-53 ASCT2 in mouse brain. J Neurochem. 2009 Jan;108(2):372-83 Witte D, Ali N, Carlson N, Younes M. Overexpression of the neutral amino acid transporter ASCT2 in human Nicklin P, Bergman P, Zhang B, Triantafellow E, Wang H, colorectal adenocarcinoma. Anticancer Res. 2002 Sep- Nyfeler B, Yang H, Hild M, Kung C, Wilson C, Myer VE, Oct;22(5):2555-7 MacKeigan JP, Porter JA, Wang YK, Cantley LC, Finan PM, Murphy LO. Bidirectional transport of amino acids Deitmer JW, Bröer A, Bröer S. Glutamine efflux from regulates mTOR and autophagy. Cell. 2009 Feb astrocytes is mediated by multiple pathways. J 6;136(3):521-34 Neurochem. 2003 Oct;87(1):127-35 Galluccio M, Pingitore P, Scalise M, Indiveri C. Cloning, Doli ńska M, Dybel A, Zabłocka B, Albrecht J. Glutamine large scale over-expression in E. coli and purification of the transport in C6 glioma cells shows ASCT2 system components of the human LAT 1 (SLC7A5) amino acid characteristics. Neurochem Int. 2003 Sep-Oct;43(4-5):501- transporter. Protein J. 2013 Aug;32(6):442-8 7 Hassanein M, Hoeksema MD, Shiota M, Qian J, Harris BK, Li R, Younes M, Frolov A, Wheeler TM, Scardino P, Ohori Chen H, Clark JE, Alborn WE, Eisenberg R, Massion PP. M, Ayala G. Expression of neutral amino acid transporter SLC1A5 mediates glutamine transport required for lung ASCT2 in human prostate. Anticancer Res. 2003 Jul- cancer cell growth and survival. Clin Cancer Res. 2013 Aug;23(4):3413-8 Feb 1;19(3):560-70 Marin M, Lavillette D, Kelly SM, Kabat D. N-linked Kim S, Kim do H, Jung WH, Koo JS. Expression of glycosylation and sequence changes in a critical negative glutamine metabolism-related proteins according to control region of the ASCT1 and ASCT2 neutral amino molecular subtype of breast cancer. Endocr Relat Cancer. acid transporters determine their retroviral receptor 2013 Jun;20(3):339-48 functions. J Virol. 2003 Mar;77(5):2936-45 Pingitore P, Pochini L, Scalise M, Galluccio M, Hedfalk K, Bungard CI, McGivan JD. Glutamine availability up- Indiveri C. Large scale production of the active human regulates expression of the amino acid transporter protein ASCT2 (SLC1A5) transporter in Pichia pastoris--functional ASCT2 in HepG2 cells and stimulates the ASCT2 and kinetic asymmetry revealed in proteoliposomes. promoter. Biochem J. 2004 Aug 15;382(Pt 1):27-32 Biochim Biophys Acta. 2013 Sep;1828(9):2238-46 Fuchs BC, Perez JC, Suetterlin JE, Chaudhry SB, Bode BP. Inducible antisense RNA targeting amino acid This article should be referenced as such: transporter ATB0/ASCT2 elicits apoptosis in human Indiveri C, Pochini L, Galluccio M, Scalise M. SLC1A5 hepatoma cells. Am J Physiol Gastrointest Liver Physiol. (solute carrier family 1 (neutral amino acid transporter), 2004 Mar;286(3):G467-78 member 5). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):673-677.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 677 Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Gene Section Short Communication

USB1 (U6 snRNA biogenesis 1) Elisa Adele Colombo Genetica Medica, Dipartimento di Scienze della Salute, Universita degli Studi di Milano, Italy (EAC)

Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/USB1ID44608ch16q21.html DOI: 10.4267/2042/54037 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract DNA/RNA C16orf57 alias USB1 is the gene which mutations Description underlie poikiloderma with neutropenia (PN) According to UCSC database (GRCh37/hg19, syndrome, a rare genodermatosis with autosomic Feb.2009), USB1 gene maps in the region between recessive inheritance. 58035277 and 58055527 bp from pter of PN patients have an increased risk to develop chromosome 16 with a centromeric-telomeric myelodysplasia and acute myeloid leukaemia in the orientation. second decade of life. It spans 20 kb and is composed of seven exons In 2012, the protein encoded by USB1 has been (GI:305855061; NM_024598.3) (Fig.2). recognised to be a 2H phosphodiesterase involved in the processing of U6 snRNA, but its action Transcription pathway and hence role in the pathogenesis of PN Two physiological isoforms, generated by has not yet been elucidated. alternative splicing (Fig. 2), have been detected in normal samples (leucocytes, keratinocytes, Identity melanocytes and fibroblasts). The major transcript of 2282 nt (isoform 1, NM_024598.3) includes all Other names: C16orf57, EC 3.1.4., hUsb1, the seven exons of the gene, while the shorter HVSL1, Mpn1, PN isoform of 1217 nt (NM_001204911.1) comprises HGNC (Hugo): USB1 the first three exons and an alternative terminal Location: 16q21 fourth exon located in IVS3 (Arnold et al., 2010).

Figure 1. The region on chromosome 16q21 containing USB1 and its neighbouring genes ZNF139 (zinc finger protein 319) and MMP15 (matrix metalloproteinase 15) (UCSC database -GRCh37/hg19, Feb 2009).

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 678 USB1 (U6 snRNA biogenesis 1) Colombo EA

Figure 2. Schematic representation of exon-intron structure of USB1 and the two major transcripts resulting from alternative splicing of the two mutually exclusive exons 4.

Several additional transcripts, a few detected in and serine residues (H120, S122, and H208, S210) cancer samples, are reported in the Ensembl which are essential for its catalytic activity. database. Recognition of these motifs by computational Pseudogene analysis of the protein sequence has predicted USB1 belongs to the 2H phosphodiesterase No pseudogene for USB1 is known. superfamily present in bacteria, archea and eukaryotes (Colombo et al., 2012). Protein The protein has a globular architecture with two juxtaposed lobes with a pseudo two-fold symmetry Description separated by a central groove, which exposes the The crystal structure of the human USB1 protein, two HLSL motifs of the active site (Fig.3). translated by isoform 1 mRNA has been recently resolved (Hilcenko et al., 2013). Expression The main USB1 protein comprises 265 aa, while USB1 is ubiquitously expressed in humans (Volpi translation of isoform 4 mRNA predicts a 186 et al., 2010). amino acid protein with a different C-terminus. The high evolutionary conservation of the protein is The USB1 protein is characterized by two consistent with the housekeeping function of the tetrapeptide motifs (HLSL), containing histidine gene.

Figure 3. Ribbon model of the USB1 protein showing its globular symmetrical conformation with two lobes separated by a central groove that exposes the catalytic site containing the two HLSL motifs (encircled). The terminal lobe comprises both the N- and the C-termini. Both the terminal and transit lobe consist of antiparallel β-sheets and α-helices (modified from Colombo et al., 2012).

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 679 USB1 (U6 snRNA biogenesis 1) Colombo EA

Localisation Mutations A nuclear localization of USB1 has been demonstrated in HeLa cells (Mroczek et al., 2012); Germinal both nuclear and mitochondrial localizations have Biallelic mutations in USB1 gene (OMIM*613276) been observed for the yeast orthologue (Glatigny et cause poikiloderma with neutropenia syndrome al., 2011). (OMIM#604173). Function To date, 19 different "loss-of-function" mutations have been identified in 38 molecularly tested PN Usb1 is a 3'-5' RNA exoribonuclease that trims the patients: 7 non-sense mutations, 6 out-of-frame 3' end of the U6 snRNA leading to the formation of deletions and 6 canonical splice site mutations. The a terminal 2',3' cyclic phosphate. This post- latter also include the only missense mutation so far transcriptionally modification influences U6 reported which however leads to exon skipping stability and recycling. Evidence has been obtained (Volpi et al., 2010). Recurrent mutations can be in yeast where Usb1 depletion leads to reduced identified in patients of Navajo, Turkish and levels of U6, generalized pre-mRNA splicing Caucasian origin attesting a founder effect defects and shorter telomeres. In human use of PN (Colombo et al., 2012). cell lines confirmed that U6 is a substrate of USB1, but failed to reveal a splicing defect leaving Somatic unsolved how PN develops (Hilcenko et al., 2012; No information is currently available on mutations Mroczek et al., 2012; Shchepachev et al., 2012). of USB1 in sporadic cancers.

Figure 4. Map across the USB1 gene of the currently known 19 mutations. Nonsense mutations are represented with a red hexagon, deletions with a yellow star and splicing mutations with a blue triangle. The Table lists for each mutation the intragenic position, the description (cDNA nomenclature) and the effect at the protein level.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 680 USB1 (U6 snRNA biogenesis 1) Colombo EA

stratify the patients according to life-long cancer Implicated in risk (myelodysplasia and solid tumours). Poikiloderma with neutropenia Further studies focussing on the alternative syndrome (PN) transcript are necessary to establish the role of isoform 4 on PN pathogenesis and prognosis. Note The disease is caused by mutations affecting the References gene represented in this entry. The clinical presentation of PN patients partially Arnold AW, Itin PH, Pigors M, Kohlhase J, Bruckner- Tuderman L, Has C. Poikiloderma with neutropenia: a overlaps that of patients affected with Rothmund- novel C16orf57 mutation and clinical diagnostic criteria. Br Thomson syndrome (RTS; OMIM#268400) and J Dermatol. 2010 Oct;163(4):866-9 dyskeratosis congenita (DC; OMIM#613987, Volpi L, Roversi G, Colombo EA, Leijsten N, Concolino D, #613988, #613989, #615190, #224230). Calabria A, Mencarelli MA, Fimiani M, Macciardi F, Pfundt Disease R, Schoenmakers EF, Larizza L. Targeted next-generation sequencing appoints c16orf57 as clericuzio-type Poikiloderma with neutropenia is a rare inherited poikiloderma with neutropenia gene. Am J Hum Genet. genodermatosis characterized by skin alterations 2010 Jan;86(1):72-6 (poikiloderma, nail dystrophy, palmo-plantar Glatigny A, Mathieu L, Herbert CJ, Dujardin G, Meunier B, hyperkeratosis), short stature and non cyclic Mucchielli-Giorgi MH. An in silico approach combined with neutropenia. in vivo experiments enables the identification of a new In infancy, neutropenia is responsible of the protein whose overexpression can compensate for specific respiratory defects in Saccharomyces cerevisiae. BMC recurrent infections, mainly of the respiratory Syst Biol. 2011 Oct 25;5:173 system, observed in PN patients and, later in life, may lead to myelodysplastic syndrome and acute Colombo EA, Bazan JF, Negri G, Gervasini C, Elcioglu NH, Yucelten D, Altunay I, Cetincelik U, Teti A, Del Fattore myeloid leukaemia. Squamous cell carcinoma has A, Luciani M, Sullivan SK, Yan AC, Volpi L, Larizza L. also been reported in PN patients. Novel C16orf57 mutations in patients with Poikiloderma To date, 38 out of 66 PN patients described in with Neutropenia: bioinformatic analysis of the protein and literature have been molecularly tested and found to predicted effects of all reported mutations. Orphanet J Rare Dis. 2012 Jan 23;7:7 carry biallelic mutations of the USB1 gene. Most of the reported patients carry homozygous mutations, Mroczek S, Krwawicz J, Kutner J, Lazniewski M, Kuciński I, Ginalski K, Dziembowski A. C16orf57, a gene mutated in attesting inheritance by descent of the same poikiloderma with neutropenia, encodes a putative ancestral mutation. phosphodiesterase responsible for the U6 snRNA 3' end Prognosis modification. Genes Dev. 2012 Sep 1;26(17):1911-25 The knowledge of USB1 3D structure with the Shchepachev V, Wischnewski H, Missiaglia E, Soneson C, essential amino acid motifs of the catalytic site Azzalin CM. Mpn1, mutated in poikiloderma with neutropenia protein 1, is a conserved 3'-to-5' RNA might enhance the prediction of USB1 mutation exonuclease processing U6 small nuclear RNA. Cell Rep. effects. 2012 Oct 25;2(4):855-65 All the mutations reported so far in PN patients (no. Hilcenko C, Simpson PJ, Finch AJ, Bowler FR, Churcher 19) interfere with USB1 function: 16 disrupt the MJ, Jin L, Packman LC, Shlien A, Campbell P, Kirwan M, catalytic activity due to the loss of one or both Dokal I, Warren AJ. Aberrant 3' oligoadenylation of HLSL motifs, while the remaining 3 mutations, spliceosomal U6 small nuclear RNA in poikiloderma with although not affecting the catalytically active neutropenia. Blood. 2013 Feb 7;121(6):1028-38 tetrapeptide motifs destroy the internal symmetry of This article should be referenced as such: the protein. Owing to the restricted number of molecularly characterised PN patients no mutation- Colombo EA. USB1 (U6 snRNA biogenesis 1). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):678-681. phenotype correlations have emerged suitable to

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Leukaemia Section Short Communication t(9;15)(p13;q24) PAX5/PML Jean-Loup Huret Genetics, Dept Medical Information, University of Poitiers, CHU Poitiers Hospital, F-86021 Poitiers, France (JLH)

Published in Atlas Database: January 2014 Online updated version : http://AtlasGeneticsOncology.org/Anomalies/t0915p13q24ID1561.html DOI: 10.4267/2042/54038 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract Genes involved and Short Communication on t(9;15)(p13;q24) proteins PAX5/PML, with data on clinics, and the genes implicated. PAX5 Location Identity 9p13.2 Protein Note 391 amino acids; from N-term to C-term, PAX5 The translocation is noted with various breakpoints contains: a paired domain (aa: 16-142); an on chromosome 15, ranging from q22 to q25 (this is octapeptide (aa: 179-186); a partial homeodomain reminiscent of the t(15;17) PML/RARA). (aa: 228-254); a transactivation domain (aa: 304- 359); and an inhibitory domain (aa: 359-391). Clinics and pathology Lineage-specific transcription factor; recognizes the concensus recognition sequence Disease GNCCANTGAAGCGTGAC, where N is any B-cell acute lymphoblastic leukemia (B-ALL) nucleotide. Involved in B-cell differentiation. Entry Epidemiology of common lymphoid progenitors into the B cell lineage depends on E2A, EBF1, and PAX5; Two cases to date, a 9-month old girl and a 1.5-year activates B-cell specific genes and repress genes old boy, both with a CD10+ ALL (Nebral et al., involved in other lineage commitments. Activates 2007; Nebral et al., 2009). the surface cell receptor CD19 and repress FLT3. Prognosis Pax5 physically interacts with the RAG1/RAG2 complex, and removes the inhibitory signal of the A patient remains in complete remission 84 months lysine-9-methylated histone H3, and induces V-to- from diagnosis, while the other one had a testicular DJ rearrangements. Genes repressed by PAX5 relapse 2 years after diagnosis and died. expression in early B cells are restored in their function in mature B cells and plasma cells, and Cytogenetics PAX5 repressed (Fuxa et al., 2004; Johnson et al., 2004; Zhang et al., 2006; Cobaleda et al., 2007; Cytogenetics morphological Medvedovic et al., 2011). The translocation was the sole abnormality.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 682 t(9;15)(p13;q24) PAX5/PML Huret JL

PAX5/PML fusion protein.

PML Fusion protein Location Description 15q24.1 1099 amino acids. The predicted fusion protein Protein contains the DNA binding paired domain of PAX5 882 amino-acids (aa) and shorter isoforms with (260 aa from PAX5) and most of PML (839 aa). distinct C terminus sequences; from N-term to C- term, PML contains: a proline rich domain (aa 3- References 46); a zinc finger (RING finger type) (aa 57-92); Grignani F, Testa U, Rogaia D, Ferrucci PF, Samoggia P, two zinc fingers (B-box types) (aa 124-166 and aa Pinto A, Aldinucci D, Gelmetti V, Fagioli M, Alcalay M, 183-236); a coiled coil made of hydrophobic aa Seeler J, Grignani F, Nicoletti I, Peschle C, Pelicci PG. Effects on differentiation by the promyelocytic leukemia heptad repeats (aa 228-253); an interaction domain PML/RARalpha protein depend on the fusion of the PML with PER2 (aa 448-555); a nuclear localization protein dimerization and RARalpha DNA binding domains. signal (aa 476-490); a proline rich domain (aa 504- EMBO J. 1996 Sep 16;15(18):4949-58 583); a serine rich domain (aa 506-540); and a Fuxa M, Skok J, Souabni A, Salvagiotto G, Roldan E, sumo interaction motif (aa 556-562). Busslinger M. Pax5 induces V-to-DJ rearrangements and The RING finger, B-boxes, and coiled-coil region locus contraction of the immunoglobulin heavy-chain gene. form a tripartite motif known as the TRIM or the Genes Dev. 2004 Feb 15;18(4):411-22 RBCC motif, and is associated with E3 ubiquitin Johnson K, Pflugh DL, Yu D, Hesslein DG, Lin KI, Bothwell ligase activity. AL, Thomas-Tikhonenko A, Schatz DG, Calame K. B cell- PML is the organizer of nuclear domains called specific loss of histone 3 lysine 9 methylation in the V(H) locus depends on Pax5. Nat Immunol. 2004 Aug;5(8):853- nuclear bodies, which recruit a wide variety of 61 proteins, most often sumoylated. PML is involved Zhang Z, Espinoza CR, Yu Z, Stephan R, He T, Williams in DNA damage response, cell division control, GS, Burrows PD, Hagman J, Feeney AJ, Cooper MD. chromosome instability, and is a clock regulator via Transcription factor Pax5 (BSAP) transactivates the RAG- regulation of PER2 expression. PML has pro- mediated V(H)-to-DJ(H) rearrangement of immunoglobulin apoptotic functions, induces senescence, inhibits genes. Nat Immunol. 2006 Jun;7(6):616-24 angiogenesis and cell migration (Grignani et al., Cobaleda C, Schebesta A, Delogu A, Busslinger M. Pax5: 1996; Chen et al., 2012; de Thé et al., 2012). the guardian of B cell identity and function. Nat Immunol. 2007 May;8(5):463-70 Result of the chromosomal Nebral K, König M, Harder L, Siebert R, Haas OA, Strehl S. Identification of PML as novel PAX5 fusion partner in anomaly childhood acute lymphoblastic leukaemia. Br J Haematol. 2007 Oct;139(2):269-74 Hybrid gene Nebral K, Denk D, Attarbaschi A, König M, Mann G, Haas Description OA, Strehl S. Incidence and diversity of PAX5 fusion genes in childhood acute lymphoblastic leukemia. Fusion of PAX5 exon 6 to PML exon 2. Leukemia. 2009 Jan;23(1):134-43

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 683 t(9;15)(p13;q24) PAX5/PML Huret JL

Medvedovic J, Ebert A, Tagoh H, Busslinger M. Pax5: a de Thé H, Le Bras M, Lallemand-Breitenbach V. The cell master regulator of B cell development and biology of disease: Acute promyelocytic leukemia, arsenic, leukemogenesis. Adv Immunol. 2011;111:179-206 and PML bodies. J Cell Biol. 2012 Jul 9;198(1):11-21

Chen RH, Lee YR, Yuan WC. The role of PML This article should be referenced as such: ubiquitination in human malignancies. J Biomed Sci. 2012 Aug 30;19:81 Huret JL. t(9;15)(p13;q24) PAX5/PML. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):682-684.

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Leukaemia Section Short Communication t(1;9)(p13;p12) PAX5/HIPK1 Jean-Loup Huret Genetics, Dept Medical Information, University of Poitiers, CHU Poitiers Hospital, F-86021 Poitiers, France (JLH)

Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Anomalies/t0109p13p12ID1557.html DOI: 10.4267/2042/54039 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

degradation of proteins) (aa 892-972), SUMO Abstract interaction motifs, required for nuclear localization Short communication on t(1;9)(p13;p12) and kinase activity (aa 902-926), a domain PAX5/HIPK1, with data on clinics, and the genes interacting with DAB2IP/MAP3K5 (aa 973-1209), implicated. a histidine-rich region (aa 1086-1154), and a tyrosine-rich region (aa 1175-1209) (YH region). Clinics and pathology The lysine residues at the sumoylation motifs are the following: K25, K317, K440, K556, and K1202 Disease (Kim et al., 1998; Li et al., 2005; Swiss-Prot). B-cell acute lymphoblastic leukemia (B-ALL) First identified as a nuclear serine/threonine kinase. Homeodomain-interacting protein kinase. HIPK1 Epidemiology positively or negatively modulate signaling Only one case to date, a 3-year old boy with a pathways controlling cell proliferation and/or CD10+ ALL (Nebral et al., 2009). apoptosis (review in Rinaldo et al., 2008). HIPK1 Prognosis and HIPK2 act cooperatively as corepressors in the transcriptional activation of angiogenic genes, The patient was noted at an intermediate risk, and including MMP10 (11q22.2) and VEGFA (6p21.1), was in complete remission 6 months after that are critical for the early stage of vascular diagnosis. development (Shang et al., 2013). HIPK1 regulates the p53 signaling pathway. Genes involved and PARK7 (also called DJ-1, 1p36.23), a protein also proteins linked to the p53 signaling pathway, is able to induce HIPK1 degradation. HIPK1 HIPK1 directly phophorylates TP53 on its serine- 15. Serine 15 phosphorylation induces a rise in Location CDKN1A (p21, 6p21.2) expression and cell cycle 1p13.2 arrest (Rey et al., 2013). Protein HIPK1 phosphorylates DAXX (6p21.32), a protein 1210 amino acids (aa). From N-term to C-term, which interacts with PML (15q24.1), the organizer contains a protein kinase domain (aa 190-518), a of nuclear bodies (Ecsedy et al., 2003), and which nucleotide binding motif (aa 196-204), a domain relocalizes from the nucleus to the cytoplasm in interacting with DAB2IP (AIP1, or AF9q34, response to stress. During glucose deprivation, a 9q33.2) (aa 518-889), a nuclear localization signal pathway involving MAP3K5 (also called ASK1, (aa 844-847), a region interacting with TP53 (aa 6q23.3), -> MAP2K4 (SEK1, 17p12) -> MAPK8 885-1093), a nuclear speckle retention signal (aa (JNK1, 10q11.22) -> HIPK1 is activated, and 887-992), (corresponding to aa 860-967 in HIPK2), DAXX is relocated in the cytoplasm (Song and a PEST domain (enriched in proline (P), glutamate Lee, 2003). (E), serine (S), and threonine (T), expedite the

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 685 t(1;9)(p13;p12) PAX5/HIPK1 Huret JL

t(1;9)(p13;p13) PAX5/HIPK1 fusion protein.

TNF (TNF-alpha, 6p21.33) induces desumoylation Protein and cytoplasm translocation of HIPK1 leading to 391 amino acids; from N-term to C-term, PAX5 apoptosis (Li et al., 2005). HIPK1 and HIPK2 bind contains: a paired domain (aa: 16-142); an HOX genes homeodomains and regulate their octapeptide (aa: 179-186); a partial homeodomain expression, as well as PAX1 (20p11.22) and PAX3 (aa: 228-254); a transactivation domain (aa: 304- (2q36.1) transcription (Isono et al., 2006). HIPK1 359); and an inhibitory domain (aa: 359-391). and HIPK2 phosphorylates EP300 (22q13.2) and Lineage-specific transcription factor; recognizes the RUNX1 (21q22.12) during embryonic concensus recognition sequence development, and Hipk1/Hipk2-deficient mice GNCCANTGAAGCGTGAC, where N is any show defective definitive hematopoiesis (Aikawa et nucleotide. al., 2006). HIPK1 phosphorylates MYB (6q23.3), a Involved in B-cell differentiation. Entry of common transcriptional activator essential for the lymphoid progenitors into the B cell lineage establishment of haemopoiesis, and causes depends on E2A, EBF1, and PAX5; activates B-cell repression of MYB activation (Matre et al., 2009). specific genes and repress genes involved in other HIPK1 interacts with DVL1 (1p36.33) and TCF3 lineage commitments. Activates the surface cell (E2A, 19p13.3) and regulates Wnt/b-catenin target receptor CD19 and repress FLT3. Pax5 physically genes during early embryonic development (Louie interacts with the RAG1/RAG2 complex, and et al., 2009). HIPK1 is highly overexpressed in removes the inhibitory signal of the lysine-9- colorectal carcinomas compared with healthy methylated histone H3, and induces V-to-DJ mucosa. The highest peak of HIPK1 expression rearrangements. Genes repressed by PAX5 occurred at early stages and decreased in latter expression in early B cells are restored in their stages. HIPK1 appeared to be induced as a defense function in mature B cells and plasma cells, and mechanism to fight against intern deregulations and PAX5 repressed (Fuxa et al., 2004; Johnson et al., stressful conditions, rather than produced by the 2004; Zhang et al., 2006; Cobaleda et al., 2007; cancer cells as an indispensable factor for tumor Medvedovic et al., 2011). evolution (Rey et al., 2013). HIPK1 is expressed only in invasive breast cancer cells with three different subcellular localization, associated with Result of the chromosomal different tumor histopathologic characteristics (Park anomaly et al., 2012). PAX5 Hybrid gene Location Description 9p13.2 Fusion of PAX5 exon 5 to HIPK1 exon 9.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 686 t(1;9)(p13;p12) PAX5/HIPK1 Huret JL

Fusion protein homeodomain-interacting protein kinases hipk1 and hipk2 in the mediation of cell growth in response to Description morphogenetic and genotoxic signals. Mol Cell Biol. 2006 751 amino acids. The predicted fusion protein Apr;26(7):2758-71 contains the DNA binding paired domain of PAX5 Zhang Z, Espinoza CR, Yu Z, Stephan R, He T, Williams and the nuclear localization signal, the region GS, Burrows PD, Hagman J, Feeney AJ, Cooper MD. interacting with TP53, the nuclear speckle retention Transcription factor Pax5 (BSAP) transactivates the RAG- mediated V(H)-to-DJ(H) rearrangement of immunoglobulin signal, the PEST domain, the SUMO interaction genes. Nat Immunol. 2006 Jun;7(6):616-24 motifs, the domain interacting with Cobaleda C, Schebesta A, Delogu A, Busslinger M. Pax5: DAB2IP/MAP3K5, and theYH region from HIPK1. the guardian of B cell identity and function. Nat Immunol. 2007 May;8(5):463-70 References Rinaldo C, Siepi F, Prodosmo A, Soddu S. HIPKs: Jack of Kim YH, Choi CY, Lee SJ, Conti MA, Kim Y. all trades in basic nuclear activities. Biochim Biophys Acta. Homeodomain-interacting protein kinases, a novel family 2008 Nov;1783(11):2124-9 of co-repressors for homeodomain transcription factors. J Louie SH, Yang XY, Conrad WH, Muster J, Angers S, Biol Chem. 1998 Oct 2;273(40):25875-9 Moon RT, Cheyette BN. Modulation of the beta-catenin Ecsedy JA, Michaelson JS, Leder P. Homeodomain- signaling pathway by the dishevelled-associated protein interacting protein kinase 1 modulates Daxx localization, Hipk1. PLoS One. 2009;4(2):e4310 phosphorylation, and transcriptional activity. Mol Cell Biol. Matre V, Nordgård O, Alm-Kristiansen AH, Ledsaak M, 2003 Feb;23(3):950-60 Gabrielsen OS. HIPK1 interacts with c-Myb and modulates Song JJ, Lee YJ. Role of the ASK1-SEK1-JNK1-HIPK1 its activity through phosphorylation. Biochem Biophys Res signal in Daxx trafficking and ASK1 oligomerization. J Biol Commun. 2009 Oct 9;388(1):150-4 Chem. 2003 Nov 21;278(47):47245-52 Nebral K, Denk D, Attarbaschi A, König M, Mann G, Haas Fuxa M, Skok J, Souabni A, Salvagiotto G, Roldan E, OA, Strehl S. Incidence and diversity of PAX5 fusion Busslinger M. Pax5 induces V-to-DJ rearrangements and genes in childhood acute lymphoblastic leukemia. locus contraction of the immunoglobulin heavy-chain gene. Leukemia. 2009 Jan;23(1):134-43 Genes Dev. 2004 Feb 15;18(4):411-22 Medvedovic J, Ebert A, Tagoh H, Busslinger M. Pax5: a Johnson K, Pflugh DL, Yu D, Hesslein DG, Lin KI, Bothwell master regulator of B cell development and AL, Thomas-Tikhonenko A, Schatz DG, Calame K. B cell- leukemogenesis. Adv Immunol. 2011;111:179-206 specific loss of histone 3 lysine 9 methylation in the V(H) Park BW, Park S, Koo JS, Kim SI, Park JM, Cho JH, Park locus depends on Pax5. Nat Immunol. 2004 Aug;5(8):853- HS. Homeodomain-interacting protein kinase 1 (HIPK1) 61 expression in breast cancer tissues. Jpn J Clin Oncol. Li X, Zhang R, Luo D, Park SJ, Wang Q, Kim Y, Min W. 2012 Dec;42(12):1138-45 Tumor necrosis factor alpha-induced desumoylation and Rey C, Soubeyran I, Mahouche I, Pedeboscq S, Bessede cytoplasmic translocation of homeodomain-interacting A, Ichas F, De Giorgi F, Lartigue L. HIPK1 drives p53 protein kinase 1 are critical for apoptosis signal-regulating activation to limit colorectal cancer cell growth. Cell Cycle. kinase 1-JNK/p38 activation. J Biol Chem. 2005 Apr 2013 Jun 15;12(12):1879-91 15;280(15):15061-70 Shang Y, Doan CN, Arnold TD, Lee S, Tang AA, Reichardt Aikawa Y, Nguyen LA, Isono K, Takakura N, Tagata Y, LF, Huang EJ. Transcriptional corepressors HIPK1 and Schmitz ML, Koseki H, Kitabayashi I. Roles of HIPK1 and HIPK2 control angiogenesis via TGF-β-TAK1-dependent HIPK2 in AML1- and p300-dependent transcription, mechanism. PLoS Biol. 2013;11(4):e1001527 hematopoiesis and blood vessel formation. EMBO J. 2006 Sep 6;25(17):3955-65 This article should be referenced as such: Isono K, Nemoto K, Li Y, Takada Y, Suzuki R, Katsuki M, Huret JL. t(1;9)(p13;p12) PAX5/HIPK1. Atlas Genet Nakagawara A, Koseki H. Overlapping roles for Cytogenet Oncol Haematol. 2014; 18(9):685-687.

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Solid Tumour Section Short Communication

Lung: t(6;12)(q22;q14.1) LRIG3/ROS1 in lung adenocarcinoma Kana Sakamoto, Yuki Togashi, Kengo Takeuchi Pathology Project for Molecular Targets, The Cancer Institute, Japanese Foundation for Cancer Research, Tokyo, Japan (KS, YT, KT)

Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Tumors/t0612q22q14inLungID6496.html DOI: 10.4267/2042/54040 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

UFT. Although not administered in this case, Abstract Crizotinib and other ALK inhibitors have been Short communication on t(6;12)(q22;q14.1) reported to be effective in lung cancers with ROS1 LRIG3/ROS1 in lung adenocarcinoma with data on translocations (Bergethon et al., 2012; Shaw et al., clinics. 2012). Clinics and pathology Prognosis With 5 years of follow-up, the patient was alive Disease without relapse. Lung adenocarcinoma Epidemiology Genes involved and ROS1 translocations are found in 0.9 to 1.7% of proteins non small cell lung carcinomas and the majority of the cases are adenocarcinoma (Bergethon et al., LRIG3 2012; Davies et al., 2012; Takeuchi et al., 2012). Location Multiple fusion partners have been identified and 12q14.1 LRIG3 is one of them. As for LRIG3-ROS1, only one case, a 57-year-old Japanese male patient, has DNA / RNA been reported to date (Takeuchi et al., 2012). Leucine-Rich Repeats And Immunoglobulin-Like Domains Protein 3. Clinics The patient had a 5 pack year of smoking history ROS1 and was diagnosed as having stage 1A lung Location adenocarcinoma. 6q22 Pathology DNA / RNA This case showed moderately differentiated C-Ros Oncogene 1, Receptor Tyrosine Kinase. micropapillary pattern. A mucinous cribriform pattern which is frequently seen in cancers with Result of the chromosomal kinase fusions was not found. This case was negative for EGFR and KRAS mutations as with anomaly the most cases harboring ROS1 gene fusions. Hybrid Gene Treatment Transcript The primary tumor was surgically removed and the LRIG3-ROS1 fusion transcript was detected. patient received post-operative chemotherapy with

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 688 Lung: t(6;12)(q22;q14.1) LRIG3/ROS1 in lung adenocarcinomaSakamoto K, et al.

A. The schematic structure of LRIG3, ROS1, and LRIG3-ROS1 proteins and the cDNA sequence around the fusion point: Exon 16 of LRIG3 fused to exon 35 of ROS1. The break point of ROS1 allows the resulting fusion protein to retain the kinase domain (red). LRIG3 contains a transmembrane domain (orange). B: RT-PCR confirmation of LRIG3-ROS1 fusion: Lane M and N represent the size standard (20-bp ladder) and the non-template control, respectively. C. Fusion FISH analysis: A fusion signal (yellow) was observed in consequence of the fusion of LRIG3 (red) and ROS1 (green).

Detection Oncogenesis A 218 bp cDNA fragment harboring the fusion The oncogenicity of LRIG3-ROS1 fusion was point can be detected with LRIG3 forward primer proven in a focus formation assay and a nude (5'-ACACAGATGAGACCAACTTGC-3') and mouse tumorigenicity assay (Takeuchi et al., 2012). ROS1 reverse primer (5'- CACTGTCACCCCTTCCTTG-3'). References Fusion Protein Bergethon K, Shaw AT, Ou SH, Katayama R, Lovly CM, McDonald NT, Massion PP, Siwak-Tapp C, Gonzalez A, Description Fang R, Mark EJ, Batten JM, Chen H, Wilner KD, Kwak The fusion protein encompasses the constitutive EL, Clark JW, Carbone DP, Ji H, Engelman JA, Mino- activation of ROS1 tyrosine kinase. Kenudson M, Pao W, Iafrate AJ. ROS1 rearrangements define a unique molecular class of lung cancers. J Clin However, the mechanism of it is largely unknown. Oncol. 2012 Mar 10;30(8):863-70 The role of LRIG3 here has not been clarified. LRIG3 protein does not contain a coiled-coil Chin LP, Soo RA, Soong R, Ou SH. Targeting ROS1 with anaplastic lymphoma kinase inhibitors: a promising domain as in the case with most of the other ROS1 therapeutic strategy for a newly defined molecular subset fusion partners (Takeuchi et al., 2012). of non-small-cell lung cancer. J Thorac Oncol. 2012 In respect of the downstream signaling, several Nov;7(11):1625-30 growth and survival signaling pathways which are Davies KD, Le AT, Theodoro MF, Skokan MC, Aisner DL, common to other receptor tyrosine kinases have Berge EM, Terracciano LM, Cappuzzo F, Incarbone M, been shown to be involved. Roncalli M, Alloisio M, Santoro A, Camidge DR, Varella- Garcia M, Doebele RC. Identifying and targeting ROS1 These include PI3K/AKT, JAK/STAT3, gene fusions in non-small cell lung cancer. Clin Cancer RAS/MAPK/ERK, VAV3, and SHP-1 and SHP-2 Res. 2012 Sep 1;18(17):4570-9 pathways (Chin et al., 2012; Davies and Doebele, Shaw AT, Camidge DR, Engelman JA, Solomon BJ, Kwak 2013). EL, Clark JW, Salgia R, Shapiro G, Bang YJ, Tan W, Tye

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 689 Lung: t(6;12)(q22;q14.1) LRIG3/ROS1 in lung adenocarcinomaSakamoto K, et al.

L, Wilner KD, Stephenson P, Varella-Garcia M, Bergethon Davies KD, Doebele RC.. Molecular pathways: ROS1 K, Iafrate AJ, Ou SHI.. Clinical activity of crizotinib in fusion proteins in cancer. Clin Cancer Res. 2013 Aug advanced non-small cell lung cancer (NSCLC) harboring 1;19(15):4040-5. doi: 10.1158/1078-0432.CCR-12-2851. ROS1 gene rearrangement. J Clin Oncol 2012;30:(suppl; Epub 2013 May 29. (REVIEW) abstr 7508). This article should be referenced as such: Takeuchi K, Soda M, Togashi Y, Suzuki R, Sakata S, Hatano S, Asaka R, Hamanaka W, Ninomiya H et al.. RET, Sakamoto K, Togashi Y, Takeuchi K. Lung: ROS1 and ALK fusions in lung cancer. Nat Med. 2012 Feb t(6;12)(q22;q14.1) LRIG3/ROS1 in lung adenocarcinoma. 12;18(3):378-81. doi: 10.1038/nm.2658. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):688- 690.

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Deep Insight Section

The tumour suppressor function of the scaffolding protein spinophilin Denis Sarrouilhe, Véronique Ladeveze Laboratoire de Physiologie Humaine, Faculte de Medecine et Pharmacie, Universite de Poitiers, 6 rue de la Miletrie, Bat D1, TSA 51115, 86073 Poitiers, Cedex 9, France (DS), Laboratoire de Genetique Moleculaire de Maladies Rares, Universite de Poitiers, UFR SFA, Pole Biologie Sante, Bat B36, TSA 51106, 86073 Poitiers, Cedex 9, France (VL)

Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Deep/SpinophilinID20133.html DOI: 10.4267/2042/54041 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract Spinophilin is a scaffolding protein with modular domains that govern its interaction with a large number of cellular proteins. The Spinophilin gene locus is localized at chromosome 17q21, a chromosomal region frequently affected by genomic instability in different human tumours. The scaffolding protein interacts with the tumour-suppressor ARF which has suggested a role for Spinophilin in cell growth. More recently, in vitro and in vivo studies demonstrated that Spinophilin is a new tumour suppressor acting via the regulation of pRb. A clear downregulation of Spinophilin is found in several human cancer types. Moreover, Spinophilin loss is associated with a poor patient prognosis in carcinoma. Currently, there are controversial findings regarding a functional relationship between Spinophilin and p53 in cell cycle regulation and in carcinogenesis. Here we present the available data regarding Spinophilin function as a tumour suppressor.

Key words Actin-Binding proteIN), and the latter was further identified as Spn (Nakanishi et al., 1997). Spn is CaSR, G protein-coupled receptor, signaling expressed ubiquitously while neurabin 1 is 1- Introduction expressed almost exclusively in neuronal cells. Spn exhibits the characteristics of scaffolding proteins Protein phosphatase 1 (PP1) is a widespread with multiple protein interaction domains (Allen et expressed phosphoSerine/phosphoThreonine PP al., 1997; Sarrouilhe et al., 2006). Scaffolding involved in many cellular processes (Ceulemans proteins link signalling enzymes, substrates and and Bollen, 2004). There are four isoforms of PP1 potential effectors (such as channels, receptors) into catalytic subunit (PP1c): PP1 α, PP1 β, PP1 γ1 and a multiprotein signalling complex that may be PP1 γ2, the latter two arising through alternative anchored to the cytoskeleton. In the years after this splicing (Sasaki et al., 1990). PP1c can form discovery, the spectrum of Spn partners and complexes with up to 50 regulatory subunits functions has expanded but has remained mostly in converting the enzyme into many different forms, the field of neurobiology (Sarrouilhe et al., 2006). which have distinct substrates specificities, Spn has been implicated in the pathophysiology of restricted subcellular locations and diverse several central nervous system (CNS) diseases, regulations (Cohen, 2002). In late 1990s, a novel among which are Parkinson's disease, schizophrenia PP1c binding protein that is a potent modulator of and mood disorders (Law et al., 2004; Brown et al., PP1 activity was characterized in rat brain and 2005). Spn is highly enriched at the synaptic named spinophilin (Spn) (Allen et al., 1997). In the membrane in dendritic spines, the site of excitatory same time, two novel actin filament-binding neurotransmission and thus may control PP1 proteins were purified from rat brain and named functions during synaptic activity (Ouimet et al., neurabin 1 and neurabin 2 (NEURal tissue-specific-

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2004). Spn regulates plasticity at the postsynaptic domains, a PSD95/DLG/zo-1 (PDZ) and three density (PSD) by targeting PP1c to α-amino-3- coiled-coil domains. Figure 2 provides a schematic hydroxy-5-methylisoxazole-4-propionic acid diagram of the main Spn structural domains. (AMPA) and N-methyl-D-aspartic acid (NMDA) In the five species of the Figure 1, the coiled-coil receptors, promoting their down regulation by region has high identity with only one variation dephosphorylation and thus regulating the detected in Cricetulus griseus . The PDZ domain, efficiency of post-synaptic glutamatergic the pentapeptide motif of PP1c -binding domain neurotransmission. Spn and neurabin1 play and the sextapeptide allowing the binding different roles in hippocampal and striatal synaptic selectivity of PP1c isoforms, present the same plasticity. Spn is involved in long-term depression identity. Moreover, the phosphoSer are conserved (LTD) but not in long-term potentiation (LTP) except the Ser-177 which is only detected in rat. whereas neurabin 1 contributes selectively to LTP Being not detected in mouse (G as in primates), but not LTD (Feng et al., 2000; Allen et al., 2006; Ser-177 is not a consequence of the rodent-specific Wu et al., 2008). In the same way, the two high substitution rate. scaffolding proteins form a functional pair of Spn has been isolated from rat brain as a protein opposing regulators that reciprocally regulate interacting with F-actin (Satoh et al., 1998). Its F- signalling intensity by some seven-transmembrane actin-binding domain determined to be amino domain receptors (Wang et al., 2007). Thus, an acids 1-154 is both necessary and sufficient to emerging notion is that Spn and neurabin 1 may mediate actin polymers binding and cross-linking. differentially affect their target proteins and Nuclear Magnetic Resonance (NMR) and circular perform quite distinctive function in cell. dichroism (CD) spectroscopy studies showed that Morphological studies have established that Spn is Spn F-actin-binding domain is intrinsically enriched at plasma membrane of cells although the unstructured and that upon binding to F-actin it protein is also expressed widely throughout the adopts a more ordered structure (a phenomenom cytoplasm (Smith et al., 1999; Richman et al., 2001; also called folding-upon-binding). Another actin Tsukada et al., 2003). Spn, which is expressed binding property, namely a F-actin pointed end partly in the nucleus in mammalian cells, interacts capping activity was recently proposed for this in vitro and in vivo with the tumor-suppressor ARF domain (Schüler and Peti, 2007). Spn, PP1c and F- (Alternative Reading Frame). Moreover, a role for actin can form a trimeric complex in vitro . Spn in cell growth was suggested, and this effect A receptor-interacting domain , located between was enhanced by the interaction between Spn and amino acids 151-444, interacts with the third ARF (Vivo et al., 2001). More recent studies intracellular loop (3i) of various seven showed that Spn is a new tumour suppressor and transmembrane domain receptors (Smith et al., that a clear downregulation of this protein is found 1999; Richman et al., 2001) such as the dopamine in several cancer types (Carnero, 2012). D2 receptor (D2R), some subtypes of the α- Furthermore, Spn loss is associated with poor adrenergic (AR) and muscarinic-acetylcholine (m- patient prognosis in carcinomas (Sarrouilhe, 2014). AchR) receptors. This review aims to outline the state of knowledge The primary PP1c-binding domain is located regarding Spn function in carcinogenesis. within residues 417-494 of Spn and this domain 2- Spinophilin structure contains a pentapeptide motif (R-K-I-H-F) between amino acids 447 and 451 that is conserved in other The primate ( homo sapiens and Callithrix jacchus ) PP1c regulatory subunits. A domain C-terminal to Spn proteins contain 815 amino acids whereas the this canonical PP1-binding motif, located within rodent Spn ( rattus norvegicus and mus musculus ) amino acids 464 and 470, is essential for PP1 have 817amino acids. These sequences are very isoform selectivity in vitro and for selective similar, with few amino acids substitutions targeting in cells (Carmody et al., 2008). compared to the human sequence in C-terminus but Recently, the 3-dimentional structure of the the N-terminus is more variable even if the PP1/Spn holoenzyme was determined. Spn is an variability is weak (Figure 1). Consequently, few unstructured protein in its unbound state that differences are observed when we compared these undergoes a folding transition upon interaction with sequences to the human one: the rat and human Spn PP1c into a single, stable conformation. The proteins share 96% sequence identity (Allen et al., scaffolding protein binds to PP1c and blocks some 1997; Vivo et al., 2001). In Cricetulus griseus, the potential substrate binding sites without altering its sequence is shorter than the others: 631amino acids. active site, then didacting substrate specificity of Gene analysis and biochemical approaches have the enzyme (Ragusa et al., 2010). A further study contributed to define in Spn a number of distinct showed that the PP1/Spn holoenzyme is dynamic in modular domains. This 130 kDa protein contains solution. one F-actin-, a receptor- and a PP1c- binding

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Figure 1. Alignment of amino acid sequences of spinophilin in different species. Blast and Align programs via UniProt site were used.

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Figure 2. Schematic drawing of spinophilin structure. The canonical protein phosphatase 1-binding domain is located within amino acids 447 and 451 in spinophilin.

The complex adopts a significant more extended signalling protein (like RGS8), guanine nucleotide conformation in solution than in the crystal exchange factors (like kalirin 7), membrane structure. This is the result of a flexible linker receptors [like the α-ARs, m-AChRs, D2R, δ- and (ramino acids 490-494) between the PP1c-binding µ-opioid receptors (OR) and cholecystokinin (CCK) and the PDZ domains. The four residue flexibility receptors], and other proteins like ions channels is likely important for Spn biological role (Ragusa [The transient receptor potential canonical (TRPC), et al., 2011). the type 2 ryanodine receptor (RYR2)], TGN38 and Spn also contains a single consensus sequence in ARF. PDZ , amino acids 494-585 (Allen et al., 1997). The Shortly after the cloning of Spn as a novel PP1c- structure of the Spn PDZ domain has been recently binding protein, another laboratory cloned this solved by NMR spectroscopy. The PDZ domain protein based on its ability to bind to F-actin (Satoh directly binds to carboxy-terminal peptides derived et al., 1998). from glutamatergic AMPA and NMDA receptors Recombinant Spn and neurabin 1 interacted with (Kelker et al., 2007). each other when co-expressed in cells. On the other Sequence analysis predicted that the carboxy- hand, recombinant Spn was shown to form terminal region of Spn (amino acids 664-814) homodimers, trimers or tetramers by interaction forms 3 coiled-coil domains. Neurabins were between coiled-coil domains. Spn homomeric observed as multimeric forms in vitro and in vivo . complexes are thought to contribute to its actin- Spn and neurabin 1 homo- and hetero-dimerize via cross-linking activity (Satoh et al., 1998). their carboxy-terminal coiled-coil domains Doublecortin (DCX) is a microtubule-associated (MacMillan et al., 1999; Oliver et al., 2002). protein that can induce microtubule polymerization Consensus sequences for phosphorylation by and stabilize microtubules filaments. several protein kinases (PK), including cAMP- Immunoprecipitation experiments with brain dependent PK (PKA), Ca 2+ /calmodulin-dependent extracts showed that Spn and DCX interact PK II (CaMKII), cyclin-dependent PK5 (Cdk5), incultured cells (Tsukada et al., 2003). extracellular-signal regulated PK (ERK) and protein In vitro assays showed that DCX also binds to and tyrosine kinases were observed in Spn. Two major bundles F-actin, suggesting that the protein cross- sites of phosphorylation for PKA (Ser-177 not links microtubules and F-actin. conserved in human, and Ser-94) and two others The distribution of DCX between the two sites for CaMKII phosphorylation (Ser-100 and cytoskeletons can be regulated by Spn and by Ser-116) were located within and near the F-actin- phosphorylation of DCX and it was proposed that binding domain of Spn. The protein is Spn could localize and enhance the binding of phosphorylated in intact cells by PKA at Ser-94 and phosphorylated DCX to F-actin (Tsukada et al., Ser-177 and by CaMKII at Ser-100 (Hsieh-Wilson 2005). et al., 2003; Grossman et al., 2004). Moreover, Several studies have shown that Spn preferentially neurabins can be phosphorylated in vitro and in binds to PP1 γ1 and PP1 α isoforms in brain extracts intact cells by Cdk5 on Ser-17 and ERK2 (MAPK1) (MacMillan et al., 1999; Terry-Lorenzo et al., 2002; on Ser-15 and Ser-205, phosphoSer-17 being Carmody et al., 2004). abundant in neuronal cells (Futter et al., 2005). GST-Spn fusion proteins containing the PP1c- Several potential tyrosine phosphorylation sites lie binding domain potently inhibit PP1 enzymatic within the coiled-coil regions, within a region activity in vitro (Allen et al., 1997; Colbran et al., adjacent to the PDZ domain and within the 2003). receptor-interacting domain. However, it was recently shown that instead of 3- The Spinophilin interactome inhibiting PP1c directly, Spn regulated enzymatic activity by directing its substrate specificity Spn interactome includes cytoskeletal molecules (Ragusa et al., 2010). (F-actin, doublecortin, neurabin 1, Spn), enzymes Spn can associate with the tyrosine phosphatase (like PP1 and CaMKII), regulator of G-protein SHP-1 and the complex modulates platelet

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activation by sequestering RGS10 and RGS18. The nervous system. Spn was identified with other sequence surrounding the phosphorylation site dendritic spines proteins as a protein partner of Y398 in Spn fits a consensus ITIM sequence TRPC5 and TRPC6 channels (Goel et al., 2005). In (I/V/L/SxY(p)xx(I/V/L) and forms a binding site cardiomyocytes, Spn targets PP1 to RYR2 via for SHP1 (Ma et al., 2012). p70 S6K is a mitogen- binding to a leucine zipper (LZ) motif of RYR2 and activated PK that regulates cell survival and a LZ motif on Spn (amino acids 300-634) causing growth. p70S6K interaction with neurabin 1 dephosphorylation and modulation of the channel (Burnett et al., 1998) and Spn was demonstrated activity (Marx et al., 2001). (Allen and Greengard, unpublished observation). TGN38 is an integral membrane protein that The interaction implicates the PDZ domain of constitutively cycles between the trans -Golgi neurabins and the carboxyl-terminal five amino network (TGN) and plasma membrane via acids of the PK. CaMKII directly and indirectly endosomal intermediates. TGN38 directly interacts associates with N- and C-terminal domains of Spn. with the coiled-coil region of Spn, preferentially Thus, Spn can target CaMKII to F-actin as well as with the dimerized proteins (Stephens and Banting, target PP1 to CaMKII (Baucum et al., 2012). 1999). Spn has been shown to interact with the Regulator of G-protein signalling (RGS) proteins nuclear protein ARF in mammalian cells. The play a crucial role in the shutting off process of G- amino acids sequence 605-726, of the coiled-coil protein-mediated responses (Ishii and Kurachi, region of Spn, seems to be involved and an intact 2003). Spn binds to different members of the RGS ARF N-terminal region (amino acids 1-65) is family (Wang et al., 2005 ; Wang et al., 2007). For necessary for this interaction (Vivo et al., 2001). example, Spn binds to through the 391-545 amino 4- Spinophilin as a tumour acids of the scaffolding protein and the 6-9 amino acids of the N-terminus of RGS8 (Fujii et al., suppressor 2008). The Spn gene locus is located on chromosome 17 at Guanine nucleotide exchange factors (GEF) position 17q21.33, a cytogenetic area frequently activate small G protein through the exchange of associated with microsatellite instability and loss of bound GDP for GTP. Several GEF were shown to heterozygosity (LOH) observed in different human interact with Spn. For example , Spn, through its tumours. This region contains a relatively high carboxy-terminus containing the PDZ and coiled- density of known (such as BRCA1 ), putative as well coil domains interacts with kalirin-7, the neuronal as several yet-unidentified candidate tumour GEF for Rac1 (Penzes et al., 2001). suppressor genes located distal to BRCA1 locus. Spn interacts with some receptors that belong to the Thus, several studies in breast and ovarian superfamily of GPCRs, mainly in the CNS. Using carcinomas have suggested the presence of an the 3i loop of the D2R , Spn has been identified as a unknown tumour suppressor gene in the area that protein that specifically associates with the receptor includes the Spn locus. However, despite these in rat hippocampal (Smith et al., 1999). The 3i preliminary genetic correlations, no in-depth loops of α2A -AR , α2B -AR , and α2C -AR subtypes analysis of the role of Spn as a tumour suppressor interact also with Spn (Richman et al., 2001). More has been made. recently, it has been shown that the α1B-AR interacts The Amancio Carnero laboratory from the Instituto with Spn in vitro (Wang et al., 2005). In the de Biomedicine de Sevilla, in Spain, have cerebellum, Spn can bind to the M1-m-AChR addressed this possibility in vitro and in vivo , in using the receptor binding domain of the three articles published in 2011. In the first study, scaffolding protein (Fujii et al., 2008). Spn can also immunohistochemical analysis of 35 human lung interact with the M2- and M3-m-AchRs but the tumours at different stages and of different binding ability to the M3-m-AChR seems to be histopathological grades showed that Spn protein is weaker than those to the M1- and M2-m-AChR absent in 20% and reduced in another 37% of (Wang et al., 2007; Kurogi et al., 2009). Moreover, tumours, compared to normal lung tissue (Molina- Spn binds to the 3i loop of CCKA and CCKB Pinelo et al., 2011). The loss of Spn expression receptors (Wang et al., 2007). The receptor binding correlated with a less differentiated phenotype, domain of Spn also associates with the 3i loop and higher grade and poor prognosis. Lower or null a conserved region of the C-terminal tails of δ- and levels of Spn also correlated with nuclear µ-OR (Fourla et al., 2012). Spn also interacts with accumulation of p53, and so to mutated p53 or loss the ionotropic NMDA and AMPA -type glutamate of its wild-type activity. Moreover, loss of Spn receptors. PDZ domain directly binds to GluR2-, increased the tumourigenic properties of p53 GluR3- (AMPA receptor) and NR1C2'-, NR2A/B- deleted- or p53 mutated-lung tumour cells. The data and NR2C/D- (NMDA receptor) derived peptides of this study showed that Spn down-regulation in (Kelker et al., 2007). lung tumours contributes to carcinogenesis in the 2+ TRPC ion channels are Ca /cation selective absence of p53. There are several mechanisms that channels that are highly expressed in the central might contribute to Spn down-regulation in

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tumours, including miRNAs overexpression. regulates the expression of p14 (ARF) (Liu et al., miRNA106*, targeting Spn, are overexpressed in a 2012). Some members of the family of e2F small subset of patients with decreased Spn levels. transcription factors are also involved in cell cycle Overexpression of miRNA106* significantly regulation; in particular E2F1 which expressions increased the tumorigenic properties of lung tumour increase induces augmentation of ARF which can cells. The results suggested that miRNA106* bind MDM2 and stabilize p53. In p53 (- / -) MEF, overexpression found in a subset of lung tumours reduced levels of Spn enhanced tumorigenic might contribute to tumorigenesis through Spn potential of the cells. Indeed, inhibition of e2F by down-regulation in the absence of p53. In a second Rb being lifted, this results in cell proliferation no study, tumour suppression by Spn was explored in longer controlled by p53. Moreover, the absence of in vivo model using genetically modified mice Spn contributes to genetic alterations during MEF (Ferrer et al., 2011b). Spn-null (-/-) mice displayed immortalization, particularly p53 mutations. These decreased survival, increased the number of results extend the observations made by the authors premalignant lesions in tissues such as the using a Spn -null mice model (Ferrer et al., 2011b). mammary ducts and early appearance of In summary, the results suggested that Spn is a new spontaneous tumours, such as lymphoma, when tumour suppressor acting via the regulation of pRb compared to WT littermates. In another series of and which function is revealed in the absence of a experiments, the presence of mutant p53 activity functional p53 (Sarrouilhe and Ladeveze, 2012). (p53R172H) in the mammary glands was evaluated This is, therefore, suggestive of partially redundant on a Spn heterozygous (+/-) or homozygous (-/-) functions in their tumour suppression properties background in mice. An increased number of (Santamaría and Malumbres, 2011). The results premalignant lesions and of mammary carcinomas also suggest that the specific outcome can be were observed in Spn heterozygous (+/-) or context-dependent. Spn loss may be beneficial by homozygous (-/-) mice when compared to WT potentiating p53 in response to acute stress, and in littermates. The results confirmed the functional contrast it can be deleterious under sustained relationship between Spn and p53 in tumorigenicity mitogenic stress (Palmero, 2011). This feature is and showed that Spn loss contributes to tumour reminiscent of NIAM (Nucleolar Interaction of progression rather than the tumour initiation. In a ARF and MDM2 protein) which acts through the third study using mouse embryonic fibroblasts same partners p53 and ARF (Tompkins et al., (MEFs), it was suggested that Spn acts as a tumour 2007). suppressor by the regulation of the stability of Another Spn-interacting molecule is DCX, an actin- PP1c α, thereby regulating its activity on pRb (the binding and microtubule-binding protein that seems phosphorylated form of the Retinoblastoma to be a tumour suppressor of glioma. When DCX is protein). This function of PP1c α has been ectopically expressed into the DCX-deficient U87 associated with the growth arrest response; the glioma cells, there is a marked suppression of the hypophosphorylated form of Rb protein being the transformed phenotype. The cells manifest a most abundant when cells are delayed in their reduced rate of growth in vitro and are arrested in growth (Ceulemans and Bollen, 2004). The ectopic the G2 phase of the cell cycle. Moreover, DCX- overexpression of Spn in immortalized MEF greatly transfected U87 glioma cells do not generate reduced tumour cell growth. Moreover, the absence tumours in immunocompromised nude rats. In of Spn (Spn(-/-) MEF) down-regulated PP1 α DCX-transfected U87 cells, phosphorylated DCX activity resulting in a high level of pRb (Ferrer et binds specifically to Spn and this interaction al., 2011a). High level of proproliferative inhibits proliferation and anchorage-independent phosphorylated Rb leads to e2F activation, a growth in glioma cells. In contrast, DCX-mediated compensatory ARF transcription, and consequently growth repression is lost in glioma cells treated p53 activation. As they regulate the cell cycle, p53 with siRNA to Spn and in HEK 293 (human and ARF are both tumour suppressors, which are embryonic kidney) Spn null cell line (Santra et al., themselves regulated by MDM2 (Mouse double 2006). DCX, Spn and PP1c were found in the same minute 2) protein shuttle between the nucleus and protein complex from mouse brain extracts cytoplasm (Kamijo et al., 1998; Pomerantz et al., (Shmueli et al., 2006). 1998). Moreover, Sherr et al. (2005) suggested for DCX-mediated growth arrest in glioma cells may the first time a p53-independent pathway via the be through inactivation of PP1 activity by ARF sumoylation. Ha et al. (2007) described ARF Spn/DCX interaction in the cytosol. Inhibition of as a melanoma tumour suppressor by inducing p53- PP1 activity is involved in two mechanistic links of independent senescence. Moreover, Du et al. (2011) reduction of glioma tumour-associated demonstrated the functional roles of ERK and p21 progressions: firstly, catastrophe in mitotic for ARF in p53-independent tumour suppression. microtubule spindle that blocks mitosis; secondly, Furthermore, in a p53-independent pathway, the depolymerization of actin that inhibits glioma cell over-expression of wild-type c-myc obviously up- invasion (Santra et al., 2009).

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Figure 3. Cellular cycle regulation by spinophilin. A. In normal cells, the presence of nuclear p53 and Spn proteins regulates cell cycle. The binding of PP1ca to Spn allows dephosphorylation of pRb, which inhibits E2F1 and thus the proliferation. Furthermore both tumour suppressors (p53 and ARF) regulate the cell cycle. The nucleolar ARF is also a partner of Spn, and regulates the cell cycle via Mdm2 and E2F1. B. In the case of colorectal carcinomas, Spn play a role in regulation of cell cycle via a p53/ARF independent pathway. One hypothesis suggested by the team of Amancio Carnero is that the Ras/Raf pathway could be implicated (Estevez-Garcia et al., 2013). This cytoplasmic pathway could be regulated by cytoplasmic Spn. K-Ras: GTPase, oncogene; B-Raf: serine/threonine protein kinase, proto-oncogene; Mek: tyrosine/threonine kinase (Mapk kinase); Mapk: mitogen-activated protein kinase.

Moreover, double transfection with DCX and Spn diagnosed after the 10-year follow-up in 85.2% reduced self-renewal in brain tumour stem cells via cases with Spn low expression and 60.9% with Spn incomplete cell cycle endomitosis (Santra et al., high expression. Death occurred in 76.5% cases 2011). But, is there relevance for Spn as a with Spn low expression and in 56.5% cases with prognostic marker in patients with cancer? Spn is Spn high expression. Overall, low Spn expression is absent in 20% and reduced in another 37% of a factor for poor prognosis in hepatocellular human lung tumors (Molina-Pinelo et al., 2011). carcinoma. In vitro experiments (human hepatoma A further analysis of Spn in human tumours shows cell line HepG2) and in vivo observations (Ki67- that Spn mRNA is lost in a percentage of renal positive tumour cells) showed that reduced Spn carcinomas and lung adenocarcinomas. A clear expression significantly correlated with a higher down-regulation of Spn was found in tumoral proliferation of liver cancer cells (Aigelsreiter et al., samples of the CNS (oligodendrogliomas, 2013). anaplastic astrocytomas, glioblastomas) when In the second study, the role of Spn was explored in compared to normal nervous samples. Furthermore, colorectal carcinoma, in which a number of lower levels of Spn mRNA correlate with higher chromosomal regions are altered (Fearon, 2011). grade of ovarian carcinoma and chronic Among them, the 17q21 is lost in a high percentage myelogenous leukemia (Carnero, 2012). Two of this carcinoma (Garcia-Patiño et al., 1998). articles published in spring 2013 associated Spn Quantitative RT-PCR analysis showed that loss with poor patient prognosis in patients with approximately 25% of colorectal carcinoma carcinoma (Sarrouilhe, 2014). The 17q tumours had a greater than 50% decrease in Spn chromosomal region is commonly impaired in mRNA levels compared with normal colonic tissue. hepatocellular carcinoma (Furge et al., 2005). In the A tissue array of human colorectal carcinomas was first study, complete loss of Spn immunoreactivity generated to confirm this result by exploring the was found in 42.3% hepatocellular carcinoma and presence of Spn protein. 70% of colorectal reduced levels were found in additional 35.6% carcinomas displayed high Spn levels (similar to cases. Quantitative RT-PCR analysis confirmed in the values observed in normal tissue), 20% showed 70% cases a significant reduced Spn mRNA intermediate levels and 10% showed no expression expression in tumour tissue compared with the of Spn. Moreover, Spn down-regulation correlated corresponding non-neoplastic tissue. miRNA106*, with a more aggressive histologic phenotype targeting Spn in lung tumours, could not be (higher Ki67-positive tumour cells) and was detected in any of the hepatocellular carcinoma associated with faster relapse and poorer survival in samples. Moreover, no correlations could be found patients with advanced stages of colorectal for the number of Spn-positive tumour cells and carcinoma. The data also suggested that Spn loss p53 or ARF staining. These results suggested a induced a chemoresistance in patients with p53-independent tumorigenic role of Spn in advanced stages of colorectal carcinoma that had hepatocellular carcinoma. Disease recurrence was received adjuvant fluoropyrimidine chemotherapy

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following surgical resection. Therefore, the Sabatini DM, Snyder SH. Neurabin is a synaptic protein identification of the levels of Spn in advanced linking p70 S6 kinase and the neuronal cytoskeleton. Proc Natl Acad Sci U S A. 1998 Jul 7;95(14):8351-6 stages of colorectal biopsies has prognostic and predictive value and might contribute to select Garcia-Patiño E, Gomendio B, Lleonart M, Silva JM, Garcia JM, Provencio M, Cubedo R, España P, Ramón y patients who could or could not benefit from Cajal S, Bonilla F. Loss of heterozygosity in the region current chemotherapy. In vitro and in vivo including the BRCA1 gene on 17q in colon cancer. Cancer experiments showed no functional relationship Genet Cytogenet. 1998 Jul 15;104(2):119-23 between Spn levels and the presence or absence of Kamijo T, Weber JD, Zambetti G, Zindy F, Roussel MF, mutated p53 in colon cancer. The authors proposed Sherr CJ. Functional and physical interactions of the ARF that this correlation is dependent on the molecular tumor suppressor with p53 and Mdm2. Proc Natl Acad Sci context of the tumour cell (Estevez-Garcia et al., U S A. 1998 Jul 7;95(14):8292-7 2013). Pomerantz J, Schreiber-Agus N, Liégeois NJ, Silverman A, 5- Discussion and perspectives Alland L, Chin L, Potes J, Chen K, Orlow I, Lee HW, We are still only at the early stage in unravelling Cordon-Cardo C, DePinho RA. The Ink4a tumor suppressor gene product, p19Arf, interacts with MDM2 and the function of Spn in cell cycle regulation. Overall, neutralizes MDM2's inhibition of p53. Cell. 1998 Mar the different studies on the tumour suppressor 20;92(6):713-23 function of Spn show two pathways of cell cycle Satoh A, Nakanishi H, Obaishi H, Wada M, Takahashi K, regulation by Spn. The first model is a pathway Satoh K, Hirao K, Nishioka H, Hata Y, Mizoguchi A, Takai dependent of p53 and ARF. Y. Neurabin-II/spinophilin. An actin filament-binding protein This pathway was previously described in several with one pdz domain localized at cadherin-based cell-cell articles where Spn interacts with different partners adhesion sites. J Biol Chem. 1998 Feb 6;273(6):3470-5 localized in the nucleus (Figure 3A). The second is MacMillan LB, Bass MA, Cheng N, Howard EF, Tamura M, a pathway independent of both molecules. As Spn Strack S, Wadzinski BE, Colbran RJ. Brain actin- associated protein phosphatase 1 holoenzymes containing is ubiquitously expressed in the cell, the first model spinophilin, neurabin, and selected catalytic subunit highlights the nuclear localization of Spn and its isoforms. J Biol Chem. 1999 Dec 10;274(50):35845-54 interaction with other nuclear proteins. Smith FD, Oxford GS, Milgram SL. Association of the D2 The second model, more hypothetical, underlines dopamine receptor third cytoplasmic loop with spinophilin, the possibility that Spn could interact with a protein phosphatase-1-interacting protein. J Biol Chem. cytoplasmic partners. The studies made on 1999 Jul 9;274(28):19894-900 colorectal carcinomas show that Spn could play a Stephens DJ, Banting G. Direct interaction of the trans- role in a pathway independent of p53/ARF. One Golgi network membrane protein, TGN38, with the F-actin hypothesis is that the Ras/Raf pathway and more binding protein, neurabin. J Biol Chem. 1999 Oct 15;274(42):30080-6 precisely K-Ras/B-Raf is implicated. This pathway, via Mek (tyrosine/threonine kinase) and Mapk Feng J, Yan Z, Ferreira A, Tomizawa K, Liauw JA, Zhuo (mitogen activated protein kinase) induces M, Allen PB, Ouimet CC, Greengard P. Spinophilin regulates the formation and function of dendritic spines. transcription factors and proliferation survical Proc Natl Acad Sci U S A. 2000 Aug 1;97(16):9287-92 (Figure 3B). Marx SO, Reiken S, Hisamatsu Y, Gaburjakova M, Further studies are needed to elucidate the Gaburjakova J, Yang YM, Rosemblit N, Marks AR. underlying mechanisms linking Spn to carcinomas Phosphorylation-dependent regulation of ryanodine and expand the prognostic and predictive value of receptors: a novel role for leucine/isoleucine zippers. J Cell the Spn expression level to other types of cancer. Biol. 2001 May 14;153(4):699-708 Penzes P, Johnson RC, Sattler R, Zhang X, Huganir RL, Kambampati V, Mains RE, Eipper BA. The neuronal Rho- References GEF Kalirin-7 interacts with PDZ domain-containing Sasaki K, Shima H, Kitagawa Y, Irino S, Sugimura T, proteins and regulates dendritic morphogenesis. Neuron. Nagao M. 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Cancer Res. 2006 Dec Identification of neurabin II as a novel doublecortin 15;66(24):11726-35 interacting protein. Mech Dev. 2003 Sep;120(9):1033-43 Sarrouilhe D, di Tommaso A, Métayé T, Ladeveze V. Carmody LC, Bauman PA, Bass MA, Mavila N, DePaoli- Spinophilin: from partners to functions. Biochimie. 2006 Roach AA, Colbran RJ. A protein phosphatase-1gamma1 Sep;88(9):1099-113 isoform selectivity determinant in dendritic spine- associated neurabin. J Biol Chem. 2004 May Shmueli A, Gdalyahu A, Sapoznik S, Sapir T, Tsukada M, 21;279(21):21714-23 Reiner O. Site-specific dephosphorylation of doublecortin (DCX) by protein phosphatase 1 (PP1). Mol Cell Neurosci. Ceulemans H, Bollen M. Functional diversity of protein 2006 May-Jun;32(1-2):15-26 phosphatase-1, a cellular economizer and reset button. Physiol Rev. 2004 Jan;84(1):1-39 Ha L, Ichikawa T, Anver M, Dickins R, Lowe S, Sharpless NE, Krimpenfort P, Depinho RA, Bennett DC, Sviderskaya Grossman SD, Futter M, Snyder GL, Allen PB, Nairn AC, EV, Merlino G. ARF functions as a melanoma tumor Greengard P, Hsieh-Wilson LC. Spinophilin is suppressor by inducing p53-independent senescence. phosphorylated by Ca2+/calmodulin-dependent protein Proc Natl Acad Sci U S A. 2007 Jun 26;104(26):10968-73 kinase II resulting in regulation of its binding to F-actin. J Neurochem. 2004 Jul;90(2):317-24 Kelker MS, Dancheck B, Ju T, Kessler RP, Hudak J, Nairn AC, Peti W. Structural basis for spinophilin-neurabin Law AJ, Weickert CS, Hyde TM, Kleinman JE, Harrison receptor interaction. Biochemistry. 2007 Mar 6;46(9):2333- PJ. Reduced spinophilin but not microtubule-associated 44 protein 2 expression in the hippocampal formation in schizophrenia and mood disorders: molecular evidence for Tompkins VS, Hagen J, Frazier AA, Lushnikova T, a pathology of dendritic spines. Am J Psychiatry. 2004 Fitzgerald MP, di Tommaso A, Ladeveze V, Domann FE, Oct;161(10):1848-55 Eischen CM, Quelle DE. A novel nuclear interactor of ARF and MDM2 (NIAM) that maintains chromosomal stability. J Ouimet CC, Katona I, Allen P, Freund TF, Greengard P. Biol Chem. 2007 Jan 12;282(2):1322-33 Cellular and subcellular distribution of spinophilin, a PP1 regulatory protein that bundles F-actin in dendritic spines. Wang X, Zeng W, Kim MS, Allen PB, Greengard P, J Comp Neurol. 2004 Nov 22;479(4):374-88 Muallem S. Spinophilin/neurabin reciprocally regulate signaling intensity by G protein-coupled receptors. EMBO Brown AM, Deutch AY, Colbran RJ. 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Wu LJ, Ren M, Wang H, Kim SS, Cao X, Zhuo M. Jul;102(7):1350-7 Neurabin contributes to hippocampal long-term potentiation and contextual fear memory. PLoS One. 2008 Baucum AJ 2nd, Strack S, Colbran RJ. Age-dependent Jan 9;3(1):e1407 targeting of protein phosphatase 1 to Ca2+/calmodulin- dependent protein kinase II by spinophilin in mouse Kurogi M, Nagatomo K, Kubo Y, Saitoh O. Effects of spinophilin on the function of RGS8 regulating signals from striatum. PLoS One. 2012;7(2):e31554 M2 and M3-mAChRs. Neuroreport. 2009 Aug Carnero A. Spinophilin: a new tumor suppressor at 17q21. 26;20(13):1134-9 Curr Mol Med. 2012 Jun;12(5):528-35 Santra M, Santra S, Roberts C, Zhang RL, Chopp M. Fourla DD, Papakonstantinou MP, Vrana SM, Georgoussi Doublecortin induces mitotic microtubule catastrophe and Z. Selective interactions of spinophilin with the C-terminal inhibits glioma cell invasion. J Neurochem. 2009 domains of the δ- and µ-opioid receptors and G proteins Jan;108(1):231-45 differentially modulate opioid receptor signaling. Cell Ragusa MJ, Dancheck B, Critton DA, Nairn AC, Page R, Signal. 2012 Dec;24(12):2315-28 Peti W. Spinophilin directs protein phosphatase 1 Liu XJ, Li FN, Jiang DD, Wang XG, Liu XP, Zhang DL, specificity by blocking substrate binding sites. Nat Struct Meng CH. [Regulation of p14(ARF) expression and Mol Biol. 2010 Apr;17(4):459-64 induction of cell apoptosis with c-myc in a p53-independent Du H, Yao W, Fang M, Wu D. ARF triggers cell G1 arrest pathway]. Zhonghua Yi Xue Za Zhi. 2012 Aug by a P53 independent ERK pathway. Mol Cell Biochem. 14;92(30):2140-3 2011 Nov;357(1-2):415-22 Ma P, Cierniewska A, Signarvic R, Cieslak M, Kong H, Fearon ER. Molecular genetics of colorectal cancer. Annu Sinnamon AJ, Neubig RR, Newman DK, Stalker TJ, Brass Rev Pathol. 2011;6:479-507 LF. A newly identified complex of spinophilin and the tyrosine phosphatase, SHP-1, modulates platelet Ferrer I, Blanco-Aparicio C, Peregrina S, Cañamero M, activation by regulating G protein-dependent signaling. Fominaya J, Cecilia Y, Lleonart M, Hernandez-Losa J, Blood. 2012 Feb 23;119(8):1935-45 Ramon y Cajal S, Carnero A. Spinophilin acts as a tumor suppressor by regulating Rb phosphorylation. Cell Cycle. Sarrouilhe D, Ladeveze V. [When the curtain goes up on 2011 Aug 15;10(16):2751-62 spinophilin's tumor suppressor function]. Med Sci (Paris). 2012 Jan;28(1):26-8 Ferrer I, Peregrino S, Cañamero M, Cecilia Y, Blanco- Aparicio C, Carnero A. Spinophilin loss contributes to Aigelsreiter A, Ress AL, Bettermann K, Schauer S, Koller tumorigenesis in vivo. Cell Cycle. 2011 Jun K, Eisner F, Kiesslich T, Stojakovic T, Samonigg H, 15;10(12):1948-55 Kornprat P, Lackner C, Haybaeck J, Pichler M. Low expression of the putative tumour suppressor spinophilin is Molina-Pinelo S, Ferrer I, Blanco-Aparicio C, Peregrino S, associated with higher proliferative activity and poor Pastor MD, Alvarez-Vega J, Suarez R, Verge M, Marin JJ, prognosis in patients with hepatocellular carcinoma. Br J Hernandez-Losa J, Ramon y Cajal S, Paz-Ares L, Carnero Cancer. 2013 May 14;108(9):1830-7 A. Down-regulation of spinophilin in lung tumours contributes to tumourigenesis. J Pathol. 2011 Estevez-Garcia P, Lopez-Calderero I, Molina-Pinelo S, Sep;225(1):73-82 Muñoz-Galvan S, Salinas A, Gomez-Izquierdo L, Lucena- Cacace A, Felipe-Abrio B, Paz-Ares L, Garcia-Carbonero Palmero I. New role for Spinophilin in tumor suppression. R, Carnero A. Spinophilin loss correlates with poor patient Cell Cycle. 2011 Oct 15;10(20):3427 prognosis in advanced stages of colon carcinoma. Clin Cancer Res. 2013 Jul 15;19(14):3925-35 Ragusa MJ, Allaire M, Nairn AC, Page R, Peti W. Flexibility in the PP1:spinophilin holoenzyme. FEBS Lett. Sarrouilhe D. [The loss of expression of spinophilin is 2011 Jan 3;585(1):36-40 associated with a bad prognosis in hepatocellular and colorectal carcinomas]. Bull Cancer. 2014 Jan 1;101(1):5-6 Santamaría D, Malumbres M. Tumor suppression by Spinophilin. Cell Cycle. 2011 Sep 1;10(17):2831-2 This article should be referenced as such: Santra M, Santra S, Buller B, Santra K, Nallani A, Chopp Sarrouilhe D, Ladeveze V. The tumour suppressor function M. Effect of doublecortin on self-renewal and differentiation of the scaffolding protein spinophilin. Atlas Genet in brain tumor stem cells. Cancer Sci. 2011 Cytogenet Oncol Haematol. 2014; 18(9):691-700.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 700 Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Case Report Section Paper co-edited with the European LeukemiaNet

Translocation t(5;6)(q33-34;q23) in an acute myelomonocytic leukemia patient Adriana Zamecnikova, Soad Al Bahar, Ramesh Pandita Kuwait Cancer Control Center, Department of Hematology, Laboratory of Cancer Genetics, Kuwait (AZ, SA, RP)

Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Reports/t0506q33q23ZamecID100076.html DOI: 10.4267/2042/54042 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract Immunophenotype Positive for CD13, CD15, CD117, CD33, MPO, Case report and literature review on translocation CD45, HLDR and dim CD34 (27%) t(5;6)(q33-34;q23) in an acute myelomonocytic Diagnosis leukemia patient. Acute myelomonocytic leukemia Clinics Survival Age and sex Date of diagnosis: 03-2013 68 years old female patient. Treatment: Chemotherapy (Daunorubicin & Previous history Cytarabine combination therapy; consolidation with No preleukemia, no previous malignancy, no inborn high dose Ara-C) condition of note, no main items. Complete remission: no Organomegaly Treatment related death: no No hepatomegaly, no splenomegaly, no enlarged Relapse: yes lymph nodes, no central nervous system Phenotype at relapse: Acute myelomonocytic involvement. leukemia Blood Status: Lost Last follow up: 11-2013 WBC: 104 X 10 9/l Survival: 8 months HB: 7.8g/dl Platelets: 57 X 10 9/l Blasts: 84% Karyotype Bone marrow: Hypercellular marrow with 87% Sample: Bone marrow, blood blasts which were PAS diffuse granular positive Culture time: 24h and SBB (Sudan Black B) positive. Banding: G-banding Cyto-Pathology Results 46,XX,t(5;6)(q33-34;q23)[25] Classification Karyotype at Relapse Cytology 46,XX,t(5;6)(q33-34;q23)[1]/46,XX,t(5;6)(q33- Acute myelomonocytic leukemia 34;q23),t(7;10)(p22;q23)[19]

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 701 Translocation t(5;6)(q33-34;q23) in an acute myelomonocytic leukemia patient Zamecnikova A, et al.

Figure 1. A. Karyotype from the time of diagnosis showing the chromosomal translocation t(5;6)(q33-34;q23). B. Fluorescence in situ hybridization studies (FISH) with XL 6q21/6q23 (Metasystem, Germany) probe showing red and green signals on both, normal and der(6) chromosomes. C. Applying the XL PDGFR probe (Metasystem, Germany) showed normal signal pattern on both normal and der(5) chromosomes, indicating that PDGFR located on 5q32-33 is not involved in the translocation. D. Hybridization with whole chromosome 6 probe (Metasystem, Germany) showing translocation of chromosome 6 sequences to der(5) chromosome.

Other molecular cytogenetics technics ALL and an associated myeloproliferative Fluorescence in situ hybridization (FISH) with LSI neoplasm and C6ORF204/PDGFRB fusion AML1-ETO, LSI MLL, LSI CBFB/inv(16), LSI (Chmielecki et al 2012). EGRI/5q31 (Abbott Molecular, Downers Grove, While in this case, the chromosomal translocation IL) and XL 6q21/6q23, XL PDGFR, whole appeared to be morphologically identical to our chromosome 6 probe Metasystem, Germany). t(5;6)(q33-34;q23), in our patient PDGFRB (5q32- 33) is not rearranged and MYB (6q23)is not Other molecular cytogenetics results translocated to chromosome 5 as in a previously Normal signal patterns for LSI AML1-ETO, LSI described case. MLL, LSI CBFB/inv(16), LSI EGRI/5q31, XL Due to the availability of tyrosine kinase inhibitors 6q21/6q23 and XL PDGFR probes. for PDGFRB rearranged disorders, our findings emphasize the importance of FISH in precise Comments characterizing of chromosome rearrangements with Chromosomal translocations involving 5q33 and 5q33-34 breakpoints, especially in suboptimal 6q23 have been reported in only one patient with T- preparations.

Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 702 Translocation t(5;6)(q33-34;q23) in an acute myelomonocytic leukemia patient Zamecnikova A, et al.

Figure 2. A. Karyotype from blood cell from the time of relapse showing the t(5;6)(q33-34;q23) and a new anomaly t(7;10)(p22;q23). B. Partial karyotypes from blood and bone marrow showing the t(5;6)(q33-34;q23).

References Chromosomes Cancer. 2012 Jan;51(1):54-65 This article should be referenced as such: Chmielecki J, Peifer M, Viale A, Hutchinson K, Giltnane J, Socci ND, Hollis CJ, Dean RS, Yenamandra A, Jagasia M, Zamecnikova A, Al Bahar S, Pandita R. Translocation Kim AS, Davé UP, Thomas RK, Pao W. Systematic screen t(5;6)(q33-34;q23) in an acute myelomonocytic leukemia for tyrosine kinase rearrangements identifies a novel patient. Atlas Genet Cytogenet Oncol Haematol. 2014; C6orf204-PDGFRB fusion in a patient with recurrent T-ALL 18(9):701-703. and an associated myeloproliferative neoplasm. Genes

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Translocation t(5;6)(q33-34;q23) in an acute myelomonocytic leukemia patient Zamecnikova A, et al.

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