DOCSLIB.ORG

A PP1-Binding Motif Present in BRCA1 Plays a Role in Its DNA Repair Function Young-Mi Yu1, Serena M

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
A PP1-Binding Motif Present in BRCA1 Plays a Role in Its DNA Repair Function Young-Mi Yu1, Serena M Int. J. Biol. Sci. 2008, 4 352 International Journal of Biological Sciences ISSN 1449-2288 www.biolsci.org 2008 4(6):352-361 © Ivyspring International Publisher. All rights reserved Research Paper A PP1-binding motif present in BRCA1 plays a role in its DNA repair function Young-Mi Yu1, Serena M. Pace1, Susan R. Allen1, Chu-Xia Deng2 and Lih-Ching Hsu1,3 1. Department of Obstetrics, Gynecology and Reproductive Sciences, School of Medicine, University of Pittsburgh, Magee-Womens Research Institute, Pittsburgh, PA15213, USA 2. Genetics of Development and Diseases Branch, National Institute of Diabetes, Digestive, and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA 3. School of Pharmacy, National Taiwan University College of Medicine, Taipei 10051, Taiwan, ROC Correspondence to: Dr. Lih-Ching Hsu, School of Pharmacy, National Taiwan University College of Medicine, 12F, No 1, Section 1, Jen-Ai Road, Taipei 10051, Taiwan. Phone: +886-2-2312-3456 ext. 88400; Fax: +886-2-2391-9098; E-mail: [email protected] Received: 2008.09.30; Accepted: 2008.10.04; Published: 2008.10.04 Protein phosphatase 1α (PP1α) regulates phosphorylation of BRCA1, which contains a PP1-binding motif 898KVTF901. Mutation of this motif greatly reduces the interaction between BRCA1 and PP1α. Here we show that mutation of the PP1-binding motif abolishes the ability of BRCA1 to enhance survival of Brca1-deficient mouse mammary tumor cells after DNA damage. The Rad51 focus formation and comet assays revealed that the DNA repair function of BRCA1 was impaired when the PP1-binding motif was mutated. Analysis of subnuclear localization of GFP-tagged BRCA1 demonstrated that mutation of the PP1-binding motif affected BRCA1 redistribution in response to DNA damage. BRCA1 is required for the formation of Rad51 subnuclear foci after DNA damage. Mutation of the PP1-binding motif in BRCA1 also affected recruitment of Rad51 to sites of DNA damage. Consistent with these findings, knockdown of PP1α in BRCA1-proficient cells by small interfering RNA also significantly reduced Rad51 focus formation induced by DNA damage. Further analysis indicated that mutation of the PP1-binding motif compromised BRCA1 activities in homologous recombination. Altogether, our data implicate that interaction with PP1α is important for BRCA1 function in DNA repair. Key words: BRCA1, protein phosphatase 1, DNA repair 1. Introduction BRCA1 is important for its function in G2/M check- point control [11] and DNA repair [12]. However, Mutations of the human breast and ovarian can- BRCA1 is primarily hypophosphorylated in mitosis cer susceptibility gene BRCA1 account for the majority and G0/G1 phases of the cell cycle [5,13], suggesting of familial breast and ovarian cancer cases. The BRCA1 that BRCA1 must undergo dephosphorylation during gene encodes a nuclear phosphoprotein of 1863 amino G2/M transition. Whether or not dephosphorylation acids (Mr ~220,000). Accumulated evidence indicates of BRCA1 is critical for its function remains unclear. that BRCA1 plays an important role in multiple cellu- Serine/threonine protein phosphatase 1 (PP1) lar functions including cell cycle checkpoint control, holoenzymes consist of catalytic subunits and regula- chromosome remodeling, transcriptional regulation, tory subunits which target the catalytic subunits to recombination, and DNA repair; therefore, is crucial distinct subcellular localizations. Together, the PP1 for maintaining genomic integrity and chromosomal complexes regulate diverse cellular functions includ- stability [1-3]. ing cell cycle progression, chromosome segregation, BRCA1 is expressed and phosphorylated in a cell cytokinesis, and cellular metabolism [14]. Recent cycle-dependent manner and is mainly phosphory- studies also indicate that PP1 is involved in check- lated in S phase [4,5]. BRCA1 is also phosphorylated point activation [15] and DNA repair [16], as well as by checkpoint kinases, including hCds1/Chk2, ATM, recovery from DNA damage checkpoint arrest [17]. and ATR, in response to DNA damage induced by We previously reported that the PP1α catalytic sub- ionizing radiation (IR), UV or chemicals. Phosphoryla- unit interacts with BRCA1 and dephosphorylates the tion of BRCA1 occurs at multiple serine residues, such protein at multiple serine residues that are phos- as S1524 and S1423 by ATM and ATR [6-9] and S988 phorylated by hCds1/Chk2, ATM and ATR [18,19]. by hCds1/Chk2 [10]. The phosphorylation state of Thus, PP1α, one of the three isoforms of the PP1 cata- Int. J. Biol. Sci. 2008, 4 353 lytic subunit (α, γ, and δ), may serve as a general USA). The mutant BRCA1 expression vectors, regulator for BRCA1 phosphorylation. pGFPBRCA1F901A and pGFPBRCA1DEL, were con- Proteins interacting with the PP1 catalytic sub- structed by replacing a 4.2 kb AflII-HindIII fragment in unit, including the majority of regulatory subunits and pGFPBRCA1 with the same fragments containing the several substrates, share a consensus PP1-binding mo- mutations from modified pBRCA1GFPF901A and tif (R/K)(V/I)xF, which binds to a hydrophobic pBRCA1GFPDEL with the BRCA1 stop codon. All groove on the surface of the PP1 catalytic subunit and plasmid clones were sequence-verified. serves as an anchor for the initial binding of these Cell culture and transfection proteins to PP1 [14]. Mutation of this consensus motif by substitution of phenylalanine with alanine or dele- A mouse mammary tumor cell line 780 (Brca1 tion of the motif greatly reduces the binding between exon 11-deleted), 293T and COS-7 cells were cultured these proteins and the PP1 catalytic subunit. We re- in DMEM supplemented with 2 mM L-glutamine and cently identified a PP1-binding motif 898KVTF901 in 10% FBS, and ES-2 cells were cultured in McCoy’s 5A human BRCA1. Mutation of this motif substantially medium supplemented with 1.5 mM L-glutamine and 10% FBS. Transfection was performed using Lipofec- reduces the interaction between BRCA1 and PP1α tamine 2000 (Invitrogen, Carlsbad, CA, USA) accord- [19]. ing to the manufacturer’s instructions. Cells were BRCA1 is implicated in the repair of DNA dou- harvested for analysis 24-48 h after transfection. Tar- ble-strand breaks (DSBs) by either homologous re- get-specific 20-25 nucleotide small interfering RNAs combination (HR) or non-homologous end-joining α (NHEJ) [1,2,20]. It has been reported that BRCA1 also (siRNAs) for human PP1 and control siRNA were participates in nucleotide-excision repair [21,22]. purchased from Santa Cruz (CA, USA) and siRNA Rad51, a homologue of bacterial RecA protein, is in- transfection was also performed using Lipofectamine volved in the repair of DSBs by HR, which is an er- 2000. ror-free DNA repair process. Expression of Rad51 is Analysis of cell cycle distribution by flow cytometry also cell cycle-dependent [23] and the protein forms Cells were harvested by trypsinization, fixed in nuclear foci in S phase [24]. BRCA2 has been shown to ice-cold methanol, and subjected to propidium iodide interact with Rad51 and regulate the assembly of staining and flow cytometric analysis as described Rad51 filaments at the sites of DNA repair. BRCA1 previously [32]. forms a complex with both Rad51 [25] and BRCA2 [26]. BRCA1 associates with Rad51 at subnuclear foci Western analysis [25]. After DNA damage, BRCA1 is phosphorylated, Cells were harvested by trypsinization and lysed relocalizes along with Rad51 to the damaged regions, in RIPA buffer supplemented with protease inhibitors. and displays large nuclear foci [27,28]. BRCA1 also Cell lysates containing ~100 μg of protein were sub- associates with the MRE11/RAD50/NBS1 complex, jected to 4-12% SDS-PAGE, followed by Western which participates in HR, NHEJ, and the DNA dam- analysis. Antibodies used were human BRCA1 anti- age response [28]. It has been demonstrated that body Ab-1 (MS110) (Calbiochem, San Diego, CA, hCds1/Chk2 phosphorylation of BRCA1 regulates the USA), mouse Brca1 M-20, PP1α C-19, Rad51 H-92 an- repair of DSBs by promoting error-free HR and sup- tibodies (Santa Cruz), and γ-tubulin antibody GTU-88 pressing error-prone NHEJ [12]. BRCA1 may also par- (Sigma, St Louis, MO, USA). Quantitative analysis was ticipate in an error-free subtype of NHEJ [29,30]. Thus, performed using Quantity One software (BioRad, BRCA1 is required for accurate DNA repair and Hercules, CA, USA). BRCA1-deficient cells are hypersensitive to DNA Cell survival assay damaging agents which induce DSBs or crosslinks of DNA strands. Here, we sought to test whether inter- Cell survival was measured by colony formation. action with PP1 was involved in the DNA repair func- One day after transfection, 500 or 1000 cells were tion of BRCA1 using Brca1∆11/∆11 mouse mammary seeded in T-25 flasks. The next day, cells were either 137 tumor cells [31]. left untreated (C) or exposed to 5 Gy of IR from a Cs source (Sheperd Mark I 30-1 Irradiator) at a dose rate 2. Materials and Methods of ~110 cGy/min. Two weeks after IR treatment, colo- Vector construction nies were fixed in cold methanol and stained with Giemsa. Colonies with at least 50 cells were then To construct pGFPBRCA1, a ~5.6 kb XhoI-HindIII scored. Relative cell survival was calculated as the BRCA1 cDNA fragment containing the human BRCA1 ratio of colony numbers in irradiated and untreated cDNA sequence (containing the stop codon) was sub- groups (IR/C). cloned into pGFP-C3 (Clontech, Mountain View, CA, Int. J. Biol. Sci. 2008, 4 354 Immunofluorescence staining for three weeks. Colonies were fixed and stained with Cells were seeded into 4-well or 8-well chamber Giemsa. XHATM-resistant colonies (at least 50 cells slides. The next day, cells were either left untreated or per colony) were quantified. The HR frequency was treated with 100 μM of 5-fluorodeoxyuridine (5-FUdR) calculated as the number of resistant colonies divided 5 or irradiated with 5 Gy of IR.
Load more

Recommended publications
  • Repositório Da Universidade De Lisboa
    UNIVERSIDADE DE LISBOA FACULDADE DE CIÊNCIAS DEPARTAMENTO DE BIOLOGIA ANIMAL TOWARDS THE IDENTIFICATION OF BIOMARKERS FOR CYSTIC FIBROSIS BY PROTEOMICS NUNO MIGUEL ANTUNES GARCIA CHARRO DOUTORAMENTO EM BIOLOGIA ESPECIALIDADE BIOLOGIA MOLECULAR 2011 ii iii iv UNIVERSIDADE DE LISBOA FACULDADE DE CIÊNCIAS DEPARTAMENTO DE BIOLOGIA ANIMAL TOWARDS THE IDENTIFICATION OF BIOMARKERS FOR CYSTIC FIBROSIS BY PROTEOMICS Tese orientada pela Doutora Deborah Penque e Professora Doutora Ana Maria Viegas Gonçalves Crespo NUNO MIGUEL ANTUNES GARCIA CHARRO DOUTORAMENTO EM BIOLOGIA (BIOLOGIA MOLECULAR) 2011 v The research described in this thesis was conducted at Laboratório de Proteómica, Departamento de Genética, Instituto Nacional de Saúde Dr. Ricardo Jorge (INSA, I.P.), Lisbon, Portugal; Clinical Proteomics Facility, University of Pittsburgh Medical Centre, Pennsylvania, USA; and Laboratory of Proteomics and Analytical Technologies, National Cancer Institute at Frederick, Maryland, USA. Work partially supported by Fundação para a Ciência e a Tecnologia (FCT), Fundo Europeu para o Desenvolvimento (FEDER) (POCI/SAU-MMO/56163/2004), FCT/Poly-Annual Funding Program and FEDER/Saúde XXI Program (Portugal). Nuno Charro is a recipient of FCT doctoral fellowship (SFRH/BD/27906/2006). vi Agradecimentos/Acknowledgements “Nothing is hidden that will not be made known; Nothing is secret that will not come to light” Desde muito pequeno, a minha vontade em querer saber mais e porquê foi sempre presença constante. Ao iniciar e no decorrer da minha (ainda) curta na investigação científica, as perguntas foram mudando, o método também e várias pessoas contribuíram para o crescimento e desenvolvimento da minha personalidade científica e pessoal. Espero não me esquecer de ninguém e, se o fizer, não é intencional; apenas falibilidade.
    [Show full text]
  • Associated 16P11.2 Deletion in Drosophila Melanogaster
    ARTICLE DOI: 10.1038/s41467-018-04882-6 OPEN Pervasive genetic interactions modulate neurodevelopmental defects of the autism- associated 16p11.2 deletion in Drosophila melanogaster Janani Iyer1, Mayanglambam Dhruba Singh1, Matthew Jensen1,2, Payal Patel 1, Lucilla Pizzo1, Emily Huber1, Haley Koerselman3, Alexis T. Weiner 1, Paola Lepanto4, Komal Vadodaria1, Alexis Kubina1, Qingyu Wang 1,2, Abigail Talbert1, Sneha Yennawar1, Jose Badano 4, J. Robert Manak3,5, Melissa M. Rolls1, Arjun Krishnan6,7 & 1234567890():,; Santhosh Girirajan 1,2,8 As opposed to syndromic CNVs caused by single genes, extensive phenotypic heterogeneity in variably-expressive CNVs complicates disease gene discovery and functional evaluation. Here, we propose a complex interaction model for pathogenicity of the autism-associated 16p11.2 deletion, where CNV genes interact with each other in conserved pathways to modulate expression of the phenotype. Using multiple quantitative methods in Drosophila RNAi lines, we identify a range of neurodevelopmental phenotypes for knockdown of indi- vidual 16p11.2 homologs in different tissues. We test 565 pairwise knockdowns in the developing eye, and identify 24 interactions between pairs of 16p11.2 homologs and 46 interactions between 16p11.2 homologs and neurodevelopmental genes that suppress or enhance cell proliferation phenotypes compared to one-hit knockdowns. These interac- tions within cell proliferation pathways are also enriched in a human brain-specific network, providing translational relevance in humans. Our study indicates a role for pervasive genetic interactions within CNVs towards cellular and developmental phenotypes. 1 Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA. 2 Bioinformatics and Genomics Program, The Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA 16802, USA.
    [Show full text]
  • Cyclin-Dependent Kinase 2 (Cdk2) Controls Phosphatase-Regulated Signaling and Function in Platelets
    bioRxiv preprint doi: https://doi.org/10.1101/2020.05.31.126953; this version posted June 28, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Cyclin-dependent kinase 2 (Cdk2) controls phosphatase-regulated signaling and function in platelets Paul R. Woods, Jr.,1,2 Brian L. Hood5, Sruti Shiva,$4 Thomas P. Conrads5, Sarah Suchko,2 Richard Steinman, 1,2,4# Departments oF Medicine1, Hillman Cancer Center2, Vascular Medicine Institute3, Department oF Molecular Pharmacology and Chemical Biology4, University of Pittsburgh School of Medicine; The Henry M. Jackson Foundation For the Advancement of Military Medicine, Inc., Inova Women’s Service Line, Inova Health System5 #Corresponding author: Richard Steinman, MD, PhD Associate Professor of Medicine and Pharmacology Associate Dean, Director Medical Scientist Training Program Director, Physician Scientist Training Program University of Pittsburgh School of Medicine 2.26f Hillman Cancer Center 5117 Centre Avenue Pittsburgh, PA 15213 USA phone: 412 6233237 fax: 412 6234840 [email protected] bioRxiv preprint doi: https://doi.org/10.1101/2020.05.31.126953; this version posted June 28, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Abstract Cell cycle regulatory molecules including cyclin-dependent kinases can be recruited into non-nuclear pathways to coordinate cell cycling with the energetic state oF the cell or with Functions such as motility.
    [Show full text]
  • SHOC2–MRAS–PP1 Complex Positively Regulates RAF Activity and Contributes to Noonan Syndrome Pathogenesis
    SHOC2–MRAS–PP1 complex positively regulates RAF activity and contributes to Noonan syndrome pathogenesis Lucy C. Younga,1, Nicole Hartiga,2, Isabel Boned del Ríoa, Sibel Saria, Benjamin Ringham-Terrya, Joshua R. Wainwrighta, Greg G. Jonesa, Frank McCormickb,3, and Pablo Rodriguez-Vicianaa,3 aUniversity College London Cancer Institute, University College London, London WC1E 6DD, United Kingdom; and bHelen Diller Family Comprehensive Cancer Center, University of California, San Francisco, CA 94158 Contributed by Frank McCormick, September 18, 2018 (sent for review November 22, 2017; reviewed by Deborah K. Morrison and Marc Therrien) Dephosphorylation of the inhibitory “S259” site on RAF kinases CRAF/RAF1 mutations are also frequently found in NS and (S259 on CRAF, S365 on BRAF) plays a key role in RAF activation. cluster around the S259 14-3-3 binding site, enhancing CRAF ac- The MRAS GTPase, a close relative of RAS oncoproteins, interacts tivity through disruption of 14-3-3 binding (8) and highlighting the with SHOC2 and protein phosphatase 1 (PP1) to form a heterotri- key role of this regulatory step in RAF–ERK pathway activation. meric holoenzyme that dephosphorylates this S259 RAF site. MRAS is a very close relative of the classical RAS oncoproteins MRAS and SHOC2 function as PP1 regulatory subunits providing (H-, N-, and KRAS, hereafter referred to collectively as “RAS”) the complex with striking specificity against RAF. MRAS also func- and shares most regulatory and effector interactions as well as tions as a targeting subunit as membrane localization is required transforming ability (9–11). However, MRAS also has specific for efficient RAF dephosphorylation and ERK pathway regulation functions of its own, and uniquely among RAS family GTPases, it in cells.
    [Show full text]
  • Pig Antibodies
    Pig Antibodies gene_name sku Entry_Name Protein_Names Organism Length Identity CDX‐2 ARP31476_P050 D0V4H7_PIG Caudal type homeobox 2 (Fragment) Sus scrofa (Pig) 147 100.00% CDX‐2 ARP31476_P050 A7MAE3_PIG Caudal type homeobox transcription factor 2 (Fragment) Sus scrofa (Pig) 75 100.00% Tnnt3 ARP51286_P050 Q75NH3_PIG Troponin T fast skeletal muscle type Sus scrofa (Pig) 271 85.00% Tnnt3 ARP51286_P050 Q75NH2_PIG Troponin T fast skeletal muscle type Sus scrofa (Pig) 266 85.00% Tnnt3 ARP51286_P050 Q75NH1_PIG Troponin T fast skeletal muscle type Sus scrofa (Pig) 260 85.00% Tnnt3 ARP51286_P050 Q75NH0_PIG Troponin T fast skeletal muscle type Sus scrofa (Pig) 250 85.00% Tnnt3 ARP51286_P050 Q75NG8_PIG Troponin T fast skeletal muscle type Sus scrofa (Pig) 266 85.00% Tnnt3 ARP51286_P050 Q75NG7_PIG Troponin T fast skeletal muscle type Sus scrofa (Pig) 260 85.00% Tnnt3 ARP51286_P050 Q75NG6_PIG Troponin T fast skeletal muscle type Sus scrofa (Pig) 250 85.00% Tnnt3 ARP51286_P050 TNNT3_PIG Troponin T, fast skeletal muscle (TnTf) Sus scrofa (Pig) 271 85.00% ORF Names:PANDA_000462 EMBL EFB13877.1OrganismAiluropod High mobility group protein B2 (High mobility group protein a melanoleuca (Giant panda) ARP31939_P050 HMGB2_PIG 2) (HMG‐2) Sus scrofa (Pig) 210 100.00% Agpat5 ARP47429_P050 B8XTR3_PIG 1‐acylglycerol‐3‐phosphate O‐acyltransferase 5 Sus scrofa (Pig) 365 85.00% irf9 ARP31200_P050 Q29390_PIG Transcriptional regulator ISGF3 gamma subunit (Fragment) Sus scrofa (Pig) 57 100.00% irf9 ARP31200_P050 Q0GFA1_PIG Interferon regulatory factor 9 Sus scrofa (Pig)
    [Show full text]
  • Supplementary Table 2
    Supplementary Table 2. Differentially Expressed Genes following Sham treatment relative to Untreated Controls Fold Change Accession Name Symbol 3 h 12 h NM_013121 CD28 antigen Cd28 12.82 BG665360 FMS-like tyrosine kinase 1 Flt1 9.63 NM_012701 Adrenergic receptor, beta 1 Adrb1 8.24 0.46 U20796 Nuclear receptor subfamily 1, group D, member 2 Nr1d2 7.22 NM_017116 Calpain 2 Capn2 6.41 BE097282 Guanine nucleotide binding protein, alpha 12 Gna12 6.21 NM_053328 Basic helix-loop-helix domain containing, class B2 Bhlhb2 5.79 NM_053831 Guanylate cyclase 2f Gucy2f 5.71 AW251703 Tumor necrosis factor receptor superfamily, member 12a Tnfrsf12a 5.57 NM_021691 Twist homolog 2 (Drosophila) Twist2 5.42 NM_133550 Fc receptor, IgE, low affinity II, alpha polypeptide Fcer2a 4.93 NM_031120 Signal sequence receptor, gamma Ssr3 4.84 NM_053544 Secreted frizzled-related protein 4 Sfrp4 4.73 NM_053910 Pleckstrin homology, Sec7 and coiled/coil domains 1 Pscd1 4.69 BE113233 Suppressor of cytokine signaling 2 Socs2 4.68 NM_053949 Potassium voltage-gated channel, subfamily H (eag- Kcnh2 4.60 related), member 2 NM_017305 Glutamate cysteine ligase, modifier subunit Gclm 4.59 NM_017309 Protein phospatase 3, regulatory subunit B, alpha Ppp3r1 4.54 isoform,type 1 NM_012765 5-hydroxytryptamine (serotonin) receptor 2C Htr2c 4.46 NM_017218 V-erb-b2 erythroblastic leukemia viral oncogene homolog Erbb3 4.42 3 (avian) AW918369 Zinc finger protein 191 Zfp191 4.38 NM_031034 Guanine nucleotide binding protein, alpha 12 Gna12 4.38 NM_017020 Interleukin 6 receptor Il6r 4.37 AJ002942
    [Show full text]
  • Allelic Deletions on Chromosome 11Q13 in Multiple Endocrine Neoplasia Type 1- Associated and Sporadic Gastrinomas and Pancreatic Endocrine Tumors
    ICANCERRESEARCH57.2238—2243.June1. 19971 Allelic Deletions on Chromosome 11q13 in Multiple Endocrine Neoplasia Type 1- associated and Sporadic Gastrinomas and Pancreatic Endocrine Tumors Larisa V. Debelenko, Zhengping Zhuang, Michael R. Emmert-Buck, Settara C. Chandrasekharappa, Pachiappan Manickam, Siradanahalli C. Guru, Stephen J. Marx, Monica C. Skarulis, Allen M. Spiegel, Francis S. Collins, Robert T. Jensen, Lance A. Liotta, and Irma A. Lubensky' Laboratory of Pathology, National Cancer institute IL V. D., 1 1, M. R. E-B., L A. L, i. A. LI, National Centerfor Human Genome Research [S. C. C., P. M., S. C. G., F. S. C.], and Branches of Metabolic Diseases (S. J. M., A. M. S.], Diabetes [M. C. S.), and Digestive Diseases [R. T. ii, National Institute of Diabetes and Digestive and Kidney Diseases, N/H. Bethesda, Maryland 20892 ABSTRACT MEN] is a tumor suppressor gene (9—11).MEN1 patients are hypoth esized to inherit a mutation in one copy of the gene, and susceptible Endocrine tumors (ETs) of pancreas and duodenum occur sporadically cells in the target organs are transformed through the inactivation of and as a part of multiple endocrine neoplasia type 1 (MEN1). The MENJ the wild-type copy of the gene, potentially occurring via point muta tumor suppressor gene has been localized to chromosome 11q13 by link age analysis but has not yet isolated. Previous alleic deletion studies in tions, deletions, or gene methylation (6, 7, 10, 11). Sporadic parathy enteropancreatic ETs suggested MENJ gene involvement in tumorigenesis roid and enteropancreatic ETs have also been described to exhibit of familial pancreatic ETs (nongastrinomas) and sporadic gastrinomas.
    [Show full text]
  • Coordinated Downregulation of Spinophilin and the Catalytic Subunits of PP1, PPP1CA/B/C, Contributes to a Worse Prognosis in Lung Cancer
    www.impactjournals.com/oncotarget/ Oncotarget, 2017, Vol. 8, (No. 62), pp: 105196-105210 Research Paper Coordinated downregulation of Spinophilin and the catalytic subunits of PP1, PPP1CA/B/C, contributes to a worse prognosis in lung cancer Eva M. Verdugo-Sivianes1,2, Lola Navas1,2, Sonia Molina-Pinelo1,2, Irene Ferrer2,3, Alvaro Quintanal-Villalonga3, Javier Peinado1,4, Jose M. Garcia-Heredia1,2,5, Blanca Felipe-Abrio1,2, Sandra Muñoz-Galvan1,2, Juan J. Marin1,2,6, Luis Montuenga2,7, Luis Paz-Ares2,3 and Amancio Carnero1,2 1Instituto de Biomedicina de Sevilla (IBIS), Hospital Universitario Virgen del Rocío, Universidad de Sevilla, Consejo Superior de Investigaciones Científicas, Sevilla, Spain 2CIBER de Cáncer, Instituto de Salud Carlos III, Pabellón 11, Planta 0, Madrid, Spain 3H120-CNIO Lung Cancer Clinical Research Unit, Instituto de Investigación Hospital 12 de Octubre and CNIO, Madrid, Spain 4Radiation Oncology Department, Hospital Universitario Virgen del Rocío, Sevilla, Spain 5Department of Vegetal Biochemistry and Molecular Biology, University of Seville, Seville, Spain 6Department of Predictive Medicine and Public Health, Universidad de Sevilla, Sevilla, Spain 7Program in Solid Tumors and Biomarkers, Center for Applied Medical Research (CIMA), Pamplona, Spain Correspondence to: Amancio Carnero, email: [email protected] Keywords: Spinophilin; PP1; biomarker; lung cancer; therapy Received: May 13, 2017 Accepted: September 03, 2017 Published: October 26, 2017 Copyright: Verdugo-Sivianes et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License 3.0 (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
    [Show full text]
  • Alterations of the Pro-Survival Bcl-2 Protein Interactome in Breast Cancer
    bioRxiv preprint doi: https://doi.org/10.1101/695379; this version posted July 12, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Alterations of the pro-survival Bcl-2 protein interactome in 2 breast cancer at the transcriptional, mutational and 3 structural level 4 5 Simon Mathis Kønig1, Vendela Rissler1, Thilde Terkelsen1, Matteo Lambrughi1, Elena 6 Papaleo1,2 * 7 1Computational Biology Laboratory, Danish Cancer Society Research Center, 8 Strandboulevarden 49, 2100, Copenhagen 9 10 2Translational Disease Systems Biology, Faculty of Health and Medical Sciences, Novo 11 Nordisk Foundation Center for Protein Research University of Copenhagen, Copenhagen, 12 Denmark 13 14 Abstract 15 16 Apoptosis is an essential defensive mechanism against tumorigenesis. Proteins of the B-cell 17 lymphoma-2 (Bcl-2) family regulates programmed cell death by the mitochondrial apoptosis 18 pathway. In response to intracellular stresses, the apoptotic balance is governed by interactions 19 of three distinct subgroups of proteins; the activator/sensitizer BH3 (Bcl-2 homology 3)-only 20 proteins, the pro-survival, and the pro-apoptotic executioner proteins. Changes in expression 21 levels, stability, and functional impairment of pro-survival proteins can lead to an imbalance 22 in tissue homeostasis. Their overexpression or hyperactivation can result in oncogenic effects. 23 Pro-survival Bcl-2 family members carry out their function by binding the BH3 short linear 24 motif of pro-apoptotic proteins in a modular way, creating a complex network of protein- 25 protein interactions.
    [Show full text]
  • Anticancer Effects of Β-Elemene in Gastric Cancer Cells and Its Potential Underlying Proteins: a Proteomic Study
    ONCOLOGY REPORTS 32: 2635-2647, 2014 Anticancer effects of β-elemene in gastric cancer cells and its potential underlying proteins: A proteomic study JUN-SONG LIU, SHI-CAI HE, ZHENG-LIANG ZHANG, RUI CHEN, LIN FAN, GUANG-LIN QIU, SHUAI CHANG, LIANG LI and XIANG-MING CHE Department of General Surgery, The First Affiliated Hospital of Medical College of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China Received July 4, 2014; Accepted August 26, 2014 DOI: 10.3892/or.2014.3490 Abstract. Gastric cancer is a common malignancy with a Introduction poor prognosis. β-elemene is a broad-spectrum anticancer drug extracted from the traditional Chinese medicinal herb Gastric cancer is the fourth most common malignancy in Curcuma wenyujin. In the present study, we investigated the the world and the second leading cause of cancer-related anticancer effects of β-elemene in gastric cancer cells and the mortality (1). At present, surgical resection remains the potential proteins involved. Human SGC7901 and MKN45 main therapeutic strategy for gastric cancer, supplemented gastric cancer cells were treated with different concentrations with perioperative chemotherapy, chemoradiotherapy and/or of β-elemene. Cell viability, clonogenic survival and apoptotic immunotherapy (2-5). However, most patients are diagnosed cell death were assessed. β-elemene inhibited viability and with advanced gastric cancer which may have progressed decreased clonogenic survival of gastric cancer cells in a beyond the curative potential of surgical operation (6,7). In dose-dependent manner. Apoptosis induction contributed to addition, previous studies have demonstrated that a consid- the anticancer effects. We then employed a proteomic method, erable proportion of patients receiving potentially curative isobaric tags for relative and absolute quantitation (iTRAQ), to resection experienced recurrences which lead to unfavorable detect the proteins altered by β-elemene.
    [Show full text]
  • Role of the Holoenzyme PP1-SPN in the Dephosphorylation of the RB Family of Tumor Suppressors During Cell Cycle
    cancers Review Role of the Holoenzyme PP1-SPN in the Dephosphorylation of the RB Family of Tumor Suppressors During Cell Cycle Eva M. Verdugo-Sivianes 1,2 and Amancio Carnero 1,2,* 1 Instituto de Biomedicina de Sevilla, IBIS, Hospital Universitario Virgen del Rocio, Consejo Superior de Investigaciones Científicas, Universidad de Sevilla, Avda. Manuel Siurot s/n, 41013 Seville, Spain; [email protected] 2 CIBERONC, Instituto de Salud Carlos III, 28029 Madrid, Spain * Correspondence: [email protected]; Tel.: +34-955-92-31-11 Simple Summary: Cell cycle progression is highly regulated by modulating the phosphorylation status of retinoblastoma (RB) family proteins. This process is controlled by a balance in the action of kinases, such as the complexes formed by cyclin-dependent kinases (CDKs) and cyclins, and phosphatases, mainly the protein phosphatase 1 (PP1). However, while the phosphorylation of the RB family has been largely studied, its dephosphorylation is less known. Recently, the PP1-Spinophilin (SPN) holoenzyme has been described as the main phosphatase responsible for the dephosphorylation of RB proteins during the G0/G1 transition and at the end of G1. Here, we describe the regulation of the phosphorylation status of RB family proteins, giving importance not only to their inactivation by phosphorylation but also to their dephosphorylation to restore the cell cycle. Abstract: Cell cycle progression is highly regulated by modulating the phosphorylation status of Citation: Verdugo-Sivianes, E.M.; the retinoblastoma protein (pRB) and the other two members of the RB family, p107 and p130. This Carnero, A. Role of the Holoenzyme process is controlled by a balance in the action of kinases, such as the complexes formed by cyclin- PP1-SPN in the Dephosphorylation of dependent kinases (CDKs) and cyclins, and phosphatases, mainly the protein phosphatase 1 (PP1).
    [Show full text]
  • SUPPLEMENTARY APPENDIX Exome Sequencing Reveals Heterogeneous Clonal Dynamics in Donor Cell Myeloid Neoplasms After Stem Cell Transplantation
    SUPPLEMENTARY APPENDIX Exome sequencing reveals heterogeneous clonal dynamics in donor cell myeloid neoplasms after stem cell transplantation Julia Suárez-González, 1,2 Juan Carlos Triviño, 3 Guiomar Bautista, 4 José Antonio García-Marco, 4 Ángela Figuera, 5 Antonio Balas, 6 José Luis Vicario, 6 Francisco José Ortuño, 7 Raúl Teruel, 7 José María Álamo, 8 Diego Carbonell, 2,9 Cristina Andrés-Zayas, 1,2 Nieves Dorado, 2,9 Gabriela Rodríguez-Macías, 9 Mi Kwon, 2,9 José Luis Díez-Martín, 2,9,10 Carolina Martínez-Laperche 2,9* and Ismael Buño 1,2,9,11* on behalf of the Spanish Group for Hematopoietic Transplantation (GETH) 1Genomics Unit, Gregorio Marañón General University Hospital, Gregorio Marañón Health Research Institute (IiSGM), Madrid; 2Gregorio Marañón Health Research Institute (IiSGM), Madrid; 3Sistemas Genómicos, Valencia; 4Department of Hematology, Puerta de Hierro General University Hospital, Madrid; 5Department of Hematology, La Princesa University Hospital, Madrid; 6Department of Histocompatibility, Madrid Blood Centre, Madrid; 7Department of Hematology and Medical Oncology Unit, IMIB-Arrixaca, Morales Meseguer General University Hospital, Murcia; 8Centro Inmunológico de Alicante - CIALAB, Alicante; 9Department of Hematology, Gregorio Marañón General University Hospital, Madrid; 10 Department of Medicine, School of Medicine, Com - plutense University of Madrid, Madrid and 11 Department of Cell Biology, School of Medicine, Complutense University of Madrid, Madrid, Spain *CM-L and IB contributed equally as co-senior authors. Correspondence:
    [Show full text]