Shieldin complex promotes DNA end-joining and counters homologous recombination in BRCA1-null cells
| Autor: | Stephen P. Jackson, Mareike Herzog, Alejandra Bruna, Luca Pellegrini, Violeta Serra, Mark J. O'Connor, Zhongwu Lai, Chloé Lescale, Jacqueline J.L. Jacobs, Fengtang Yang, Jonathan Lam, Matylda Sczaniecka-Clift, Abigail Shea, Carlos Caldas, Matthias Ostermaier, Gabriel Balmus, Julia Coates, Wenming Wei, Inge de Krijger, Yaron Galanty, Mukerrem Demir, Ludovic Deriano, Petra Beli, Domenic Pilger, Harveer Dev, Rimma Belotserkovskaya, Alistair Martin, Beiyuan Fu, Ting-Wei Will Chiang, Qian Wu |
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| Přispěvatelé: | Dev, Harveer [0000-0003-2874-6894], Yang, Fengtang [0000-0002-3573-2354], Balmus, Gabriel [0000-0003-2872-4468], Serra, Violeta [0000-0001-6620-1065], Beli, Petra [0000-0001-9507-9820], Pellegrini, Luca [0000-0002-9300-497X], Deriano, Ludovic [0000-0002-9673-9525], Jacobs, Jacqueline JL [0000-0002-7704-4795], Jackson, Stephen P [0000-0001-9317-7937], Apollo - University of Cambridge Repository, Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge [UK] (CAM), Department of Biochemistry [Cambridge], Cambridge University Hospitals - NHS (CUH), Intégrité du génome, immunité et cancer - Genome integrity, Immunity and Cancer, Institut Pasteur [Paris] (IP), Netherlands Cancer Institute (NKI), Antoni van Leeuwenhoek Hospital, Cancer Research UK Cambridge Institute [Cambridge, Royaume-Uni] (CRUK), Institute of Molecular Biology (IMB), Johannes Gutenberg - Universität Mainz = Johannes Gutenberg University (JGU), Wellcome Trust Sanger Institute [Hinxton, UK], AstraZeneca US [Waltham, USA], AstraZeneca [Cambridge, UK], The Wellcome Trust Sanger Institute [Cambridge], Vall d'Hebron Institute of Oncology [Barcelone] (VHIO), Vall d'Hebron University Hospital [Barcelona], The SPJ lab is largely funded by a Cancer Research UK (CRUK) Program Grant, C6/A18796, and a Wellcome Trust (WT) Investigator Award, 206388/Z/17/Z. Core infrastructure funding was provided by CRUK grant C6946/A24843 and WT grant WT203144. S.P.J. receives a salary from the University of Cambridge. H.D. is funded by WT Clinical Fellowship 206721/Z/17/Z. TWC was supported by a Cambridge International Scholarship. D.P. is funded by Cancer Research UK studentship C6/A21454. The P.B. lab is supported by the Emmy Noether Program (BE 5342/1-1) from the German Research Foundation and a Marie Curie Career Integration Grant from the European Commission (630763). The L.P. lab is funded by the WT (investigator award 104641/Z/14/Z) and the Medical Research Council (project grant MR/N000161/1). The C.C. lab was supported with funding from CRUK. The J.J. lab was supported by the European Research Council grant ERC-StG 311565, The Dutch Cancer Society (KWF) grant KWF 10999, and the Netherlands Organization for Scientific Research (NWO) as part of the National Roadmap Large-scale Research Facilities of the Netherlands, Proteins@Work (project no. 184.032.201 to the Proteomics Facility of the Netherlands Cancer Institute). The L.D. lab is funded by the Institut Pasteur, the Institut National du Cancer (no. PLBIO16-181) and the European Research Council (starting grant agreement no. 310917). W.W. is part of the Pasteur–Paris University (PPU) International PhD program and this project received funding from the CNBG company, China. Q.W. is funded by the Wellcome Trust (200814/Z/16/Z ). The V.S. lab work was funded by the Instituto de Salud Carlos III (ISCIII), an initiative of the Spanish Ministry of Economy and Innovation partially supported by European Regional Development FEDER Funds (PI17-01080 to VS), the European Research Area-NET, Transcan-2 (AC15/00063), a non-commercial research agreement with AstraZeneca UK, and structural funds from the Agència de Gestió d’Ajuts Universitaris i de Recerca (AGAUR, 2017 SGR 540) and the Orozco Family. V.S. received a salary and travel support to C.C.’s lab from ISCIII (CP14/00228, MV15/00041) and the FERO Foundation., The authors thank all S.P.J. laboratory members for support and advice, and Cambridge colleagues N. Lawrence for OMX super-resolution microscopy support and R. Butler for help with computational image analyses and programming. The authors also thank S. Selivanova and S. Hough for help with plasmid amplification, sample preparation and tissue culture maintenance, K. Dry for extensive editorial assistance, F. Muñoz-Martinez for assistance with CRISPR–Cas9 knockout generation, L. Radu for assistance with protein purification, C. Lord (Institute of Cancer Research, London) for SUM149PT cells, D. Durocher (University of Toronto, Canada) for U2OS LacSceIII cells, F. Alt (Harvard University, USA) for CH12F3 cells and 53bp1 knockout CH12F3 cell clones, T. Honjo (Kyoto University, Japan) for permission to use the CH12F3 cell line, and J. Serrat in the Jacobs lab for technical assistance, Institut Pasteur [Paris], Johannes Gutenberg - Universität Mainz (JGU) |
| Rok vydání: | 2018 |
| Předmět: |
MESH: DNA Breaks
Double-Stranded RAD51 Cell Cycle Proteins Poly (ADP-Ribose) Polymerase Inhibitor MESH: Recombinational DNA Repair Mice MESH: Animals DNA Breaks Double-Stranded skin and connective tissue diseases Cancer Telomere-binding protein Ovarian Neoplasms MESH: Breast Neoplasms / metabolism MESH: Telomere-Binding Proteins / metabolism 3. Good health Cell biology MESH: HEK293 Cells MESH: Proteins / genetics MESH: Telomere-Binding Proteins / genetics MESH: Tumor Suppressor p53-Binding Protein 1 / metabolism MESH: Xenograft Model Antitumor Assays Telomere-Binding Proteins MESH: Ovarian Neoplasms / drug therapy Bone Neoplasms MESH: Ovarian Neoplasms / metabolism Article 03 medical and health sciences MESH: Cell Cycle Proteins MESH: Bone Neoplasms / metabolism Humans MESH: Osteosarcoma / metabolism [SDV.GEN]Life Sciences [q-bio]/Genetics MESH: Humans MESH: Tumor Suppressor p53-Binding Protein 1 / genetics Dose-Response Relationship Drug HEK 293 cells Proteins [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry Molecular Biology/Molecular biology DNA MESH: BRCA1 Protein / deficiency 030104 developmental biology Multiprotein Complexes MESH: Mad2 Proteins / metabolism MESH: Breast Neoplasms / genetics MESH: Bone Neoplasms / drug therapy Cisplatin Homologous recombination MESH: Osteosarcoma / genetics MESH: Female 0301 basic medicine DNA End-Joining Repair MESH: Proteins / metabolism MESH: Dose-Response Relationship Drug chemistry.chemical_compound MESH: Osteosarcoma / pathology MESH: Breast Neoplasms / pathology Homologous Recombination Polymerase MESH: Breast Neoplasms / drug therapy Osteosarcoma biology Chemistry BRCA1 Protein DNA damage and repair MESH: Poly(ADP-ribose) Polymerase Inhibitors / pharmacology MESH: Bone Neoplasms / genetics DNA-Binding Proteins MESH: Bone Neoplasms / pathology Mad2 Proteins Female MESH: Ovarian Neoplasms / genetics Tumor Suppressor p53-Binding Protein 1 MESH: Cisplatin / pharmacology MESH: Cell Line Tumor Lymphocytes Null [SDV.CAN]Life Sciences [q-bio]/Cancer Breast Neoplasms [SDV.BC]Life Sciences [q-bio]/Cellular Biology MESH: BRCA1 Protein / genetics Poly(ADP-ribose) Polymerase Inhibitors Cell Line Tumor MESH: Drug Resistance Neoplasm* / genetics MESH: Mad2 Proteins / genetics MESH: Ovarian Neoplasms / pathology Animals MESH: Mice MESH: Osteosarcoma / drug therapy Oligonucleotide Protective Devices Recombinational DNA Repair Cell Biology MESH: Multiprotein Complexes Xenograft Model Antitumor Assays HEK293 Cells Drug Resistance Neoplasm biology.protein MESH: DNA End-Joining Repair MESH: DNA-Binding Proteins |
| Zdroj: | Nature Cell Biology Nature Cell Biology, 2018, 20 (8), pp.954-965. ⟨10.1038/s41556-018-0140-1⟩ Nature cell biology Nature Cell Biology, Nature Publishing Group, 2018, 20 (8), pp.954-965. ⟨10.1038/s41556-018-0140-1⟩ |
| ISSN: | 1465-7392 1476-4679 |
| DOI: | 10.1038/s41556-018-0140-1⟩ |
| Popis: | International audience; BRCA1 deficiencies cause breast, ovarian, prostate and other cancers, and render tumours hypersensitive to poly(ADP-ribose) polymerase (PARP) inhibitors. To understand the resistance mechanisms, we conducted whole-genome CRISPR-Cas9 synthetic-viability/resistance screens in BRCA1-deficient breast cancer cells treated with PARP inhibitors. We identified two previously uncharacterized proteins, C20orf196 and FAM35A, whose inactivation confers strong PARP-inhibitor resistance. Mechanistically, we show that C20orf196 and FAM35A form a complex, 'Shieldin' (SHLD1/2), with FAM35A interacting with single-stranded DNA through its C-terminal oligonucleotide/oligosaccharide-binding fold region. We establish that Shieldin acts as the downstream effector of 53BP1/RIF1/MAD2L2 to promote DNA double-strand break (DSB) end-joining by restricting DSB resection and to counteract homologous recombination by antagonizing BRCA2/RAD51 loading in BRCA1-deficient cells. Notably, Shieldin inactivation further sensitizes BRCA1-deficient cells to cisplatin, suggesting how defining the SHLD1/2 status of BRCA1-deficient tumours might aid patient stratification and yield new treatment opportunities. Highlighting this potential, we document reduced SHLD1/2 expression in human breast cancers displaying intrinsic or acquired PARP-inhibitor resistance. |
| Databáze: | OpenAIRE |
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