Error-free repair of DNA double-strand breaks (DSBs) is achieved by homologous recombination (HR), and BRCA1 is an important factor for this repair pathway. In the absence of BRCA1-mediated HR, the administration of PARP inhibitors induces synthetic lethality of tumour cells of patients with breast or ovarian cancers. Despite the benefit of this tailored therapy, drug resistance can occur by HR restoration. Genetic reversion of BRCA1-inactivating mutations can be the underlying mechanism of drug resistance, but this does not explain resistance in all cases. In particular, little is known about BRCA1-independent restoration of HR. Here we show that loss of REV7 (also known as MAD2L2) in mouse and human cell lines re-establishes CTIP-dependent end resection of DSBs in BRCA1-deficient cells, leading to HR restoration and PARP inhibitor resistance, which is reversed by ATM kinase inhibition. REV7 is recruited to DSBs in a manner dependent on the H2AX-MDC1-RNF8-RNF168-53BP1 chromatin pathway, and seems to block HR and promote end joining in addition to its regulatory role in DNA damage tolerance. Finally, we establish that REV7 blocks DSB resection to promote non-homologous end-joining during immunoglobulin class switch recombination. Our results reveal an unexpected crucial function of REV7 downstream of 53BP1 in coordinating pathological DSB repair pathway choices in BRCA1-deficient cells.

Danish Cancer Society Research Center Copenhagen Denmark

Deep Sequencing Core Facility The Netherlands Cancer Institute Plesmanlaan 121 1066CX Amsterdam The Netherlands

Department of Experimental Radiation Oncology University of Texas MD Anderson Cancer Center Houston TX 77030 USA

Department of Genetics Erasmus University Medical Center Rotterdam The Netherlands

Division of Cell Biology The Netherlands Cancer Institute Plesmanlaan 121 1066CX Amsterdam The Netherlands

Division of Molecular Carcinogenesis The Netherlands Cancer Institute Plesmanlaan 121 1066CX Amsterdam The Netherlands

Division of Molecular Oncology The Netherlands Cancer Institute Plesmanlaan 121 1066CX Amsterdam The Netherlands

Division of Molecular Pathology The Netherlands Cancer Institute Plesmanlaan 121 1066CX Amsterdam The Netherlands

DNA Damage Response Laboratory London Research Institute Cancer Research UK Clare Hall South Mimms EN6 3LD UK

Institute of Animal Pathology Vetsuisse Faculty University of Bern Laengassstrasse 122 3012 Bern Switzerland

Institute of Molecular and Translational Medicine Faculty of Medicine and Dentistry Palacky University Olomouc Czech Republic

Protein Facility The Netherlands Cancer Institute Plesmanlaan 121 1066CX Amsterdam The Netherlands

The Wellcome Trust Centre for Human Genetics Roosevelt Drive Oxford OX3 7BN United Kingdom

EMBO J. 2015 Jun 12;34(12):1609-11. doi: 10.15252/embj.201591697. PubMed

Zobrazit více v PubMed

Roy R, Chun J, Powell SN. BRCA1 and BRCA2: different roles in a common pathway of genome protection. Nat. Rev. Cancer. 2012;12:68–78. PubMed PMC

Lee J-M, Ledermann JA, Kohn EC. PARP Inhibitors for BRCA1/2 mutation-associated and BRCA-like malignancies. Ann. Oncol. Off. J. Eur. Soc. Med. Oncol. ESMO. 2013 doi:10.1093/annonc/mdt384. PubMed PMC

Farmer H, et al. Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy. Nature. 2005;434:917–921. PubMed

Lord CJ, Ashworth A. Mechanisms of resistance to therapies targeting BRCA-mutant cancers. Nat. Med. 2013;19:1381–1388. PubMed

Ang JE, et al. Efficacy of chemotherapy in BRCA1/2 mutation carrier ovarian cancer in the setting of PARP inhibitor resistance: a multi-institutional study. Clin. Cancer Res. Off. J. Am. Assoc. Cancer Res. 2013;19:5485–5493. PubMed

Chun AC-S, Kok K-H, Jin D-Y. REV7 is required for anaphase-promoting complex-dependent ubiquitination and degradation of translesion DNA polymerase REV1. Cell Cycle Georget. Tex. 2013;12:365–378. PubMed PMC

Jaspers JE, et al. Loss of 53BP1 Causes PARP Inhibitor Resistance in Brca1-Mutated Mouse Mammary Tumors. Cancer Discov. 2013;3:68–81. PubMed PMC

Listovsky T, Sale JE. Sequestration of CDH1 by MAD2L2 prevents premature APC/C activation prior to anaphase onset. J. Cell Biol. 2013;203:87–100. PubMed PMC

Gan GN, Wittschieben JP, Wittschieben BØ, Wood RD. DNA polymerase zeta (pol zeta) in higher eukaryotes. Cell Res. 2008;18:174–183. PubMed

Hara K, et al. Crystal structure of human REV7 in complex with a human REV3 fragment and structural implication of the interaction between DNA polymerase zeta and REV1. J. Biol. Chem. 2010;285:12299–12307. PubMed PMC

Khalaj M, et al. A missense mutation in Rev7 disrupts formation of Polζ, impairing mouse development and repair of genotoxic agent-induced DNA lesions. J. Biol. Chem. 2014;289:3811–3824. PubMed PMC

Stap J, et al. Induction of linear tracks of DNA double-strand breaks by alpha-particle irradiation of cells. Nat. Methods. 2008;5:261–266. PubMed

Fanning E, Klimovich V, Nager AR. A dynamic model for replication protein A (RPA) function in DNA processing pathways. Nucleic Acids Res. 2006;34:4126–4137. PubMed PMC

Sartori AA, et al. Human CtIP promotes DNA end resection. Nature. 2007;450:509–514. PubMed PMC

Bouwman P, et al. 53BP1 loss rescues BRCA1 deficiency and is associated with triple-negative and BRCA-mutated breast cancers. Nat. Struct. Mol. Biol. 2010;17:688–695. PubMed PMC

Pierce AJ, Johnson RD, Thompson LH, Jasin M. XRCC3 promotes homology-directed repair of DNA damage in mammalian cells. Genes Dev. 1999;13:2633–2638. PubMed PMC

Bunting SF, et al. 53BP1 inhibits homologous recombination in Brca1-deficient cells by blocking resection of DNA breaks. Cell. 2010;141:243–254. PubMed PMC

Zimmermann M, de Lange T. 53BP1: pro choice in DNA repair. Trends Cell Biol. 2014;24:108–117. PubMed PMC

Chapman JR, et al. RIF1 is essential for 53BP1-dependent nonhomologous end joining and suppression of DNA double-strand break resection. Mol. Cell. 2013;49:858–871. PubMed PMC

Muramatsu M, et al. Class switch recombination and hypermutation require activation-induced cytidine deaminase (AID), a potential RNA editing enzyme. Cell. 2000;102:553–563. PubMed

Schenten D, et al. Pol zeta ablation in B cells impairs the germinal center reaction, class switch recombination, DNA break repair, and genome stability. J. Exp. Med. 2009;206:477–490. PubMed PMC

Kikuchi S, Hara K, Shimizu T, Sato M, Hashimoto H. Structural basis of recruitment of DNA polymerase ζ by interaction between REV1 and REV7 proteins. J. Biol. Chem. 2012;287:33847–33852. PubMed PMC

Evers B, et al. Selective inhibition of BRCA2-deficient mammary tumor cell growth by AZD2281 and cisplatin. Clin. Cancer Res. 2008;14:3916–3925. PubMed

Fradet-Turcotte A, et al. 53BP1 is a reader of the DNA-damage-induced H2A Lys 15 ubiquitin mark. Nature. 2013;499:50–54. PubMed PMC

Watanabe S, et al. JMJD1C demethylates MDC1 to regulate the RNF8 and BRCA1-mediated chromatin response to DNA breaks. Nat. Struct. Mol. Biol. 2013;20:1425–1433. PubMed

Doil C, et al. RNF168 binds and amplifies ubiquitin conjugates on damaged chromosomes to allow accumulation of repair proteins. Cell. 2009;136:435–446. PubMed

Stewart GS, et al. The RIDDLE syndrome protein mediates a ubiquitin-dependent signaling cascade at sites of DNA damage. Cell. 2009;136:420–434. PubMed

Stap J, et al. Induction of linear tracks of DNA double-strand breaks by alpha-particle irradiation of cells. Nat. Methods. 2008;5:261–266. PubMed

Bouwman P, et al. A high-throughput functional complementation assay for classification of BRCA1 missense variants. Cancer Discov. 2013;3:1142–1155. PubMed

Bartkova J, et al. DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis. Nature. 2005;434:864–870. PubMed

RAD51 recruitment but not replication fork stability associates with PARP inhibitor response in ovarian cancer patient-derived xenograft models

DNA damage signalling barrier, oxidative stress and treatment-relevant DNA repair factor alterations during progression of human prostate cancer

Cells and Stripes: A novel quantitative photo-manipulation technique