Shieldin complex promotes DNA end-joining and counters homologous recombination in BRCA1-null cells

Harveer Dev; Ting-Wei Will Chiang; Chloe Lescale; Inge de Krijger; Alistair G Martin; Domenic Pilger; Julia Coates; Matylda Sczaniecka-Clift; Wenming Wei; Matthias Ostermaier; Mareike Herzog; Jonathan Lam; Abigail Shea; Mukerrem Demir; Qian Wu; Fengtang Yang; Beiyuan Fu; Zhongwu Lai; Gabriel Balmus; Rimma Belotserkovskaya; Violeta Serra; Mark J O'Connor; Alejandra Bruna; Petra Beli; Luca Pellegrini; Carlos Caldas; Ludovic Deriano; Jacqueline J L Jacobs; Yaron Galanty; Stephen P Jackson

doi:10.1038/s41556-018-0140-1

Shieldin complex promotes DNA end-joining and counters homologous recombination in BRCA1-null cells

Nat Cell Biol. 2018 Aug;20(8):954-965. doi: 10.1038/s41556-018-0140-1. Epub 2018 Jul 18.

Authors

Harveer Dev^{1

2}, Ting-Wei Will Chiang^#¹, Chloe Lescale^#³, Inge de Krijger^#⁴, Alistair G Martin⁵, Domenic Pilger¹, Julia Coates¹, Matylda Sczaniecka-Clift¹, Wenming Wei³, Matthias Ostermaier⁶, Mareike Herzog¹, Jonathan Lam¹, Abigail Shea⁵, Mukerrem Demir¹, Qian Wu⁷, Fengtang Yang⁸, Beiyuan Fu⁸, Zhongwu Lai⁹, Gabriel Balmus^{1

8}, Rimma Belotserkovskaya¹, Violeta Serra¹⁰, Mark J O'Connor¹¹, Alejandra Bruna⁵, Petra Beli⁶, Luca Pellegrini⁷, Carlos Caldas⁵, Ludovic Deriano¹², Jacqueline J L Jacobs¹³, Yaron Galanty¹⁴, Stephen P Jackson¹⁵

Affiliations

¹ The Wellcome Trust/Cancer Research UK Gurdon Institute and Department of Biochemistry, University of Cambridge, Cambridge, UK.
² Academic Urology Group, Department of Surgery, Cambridge University Hospitals NHS Foundation Trust, Addenbrooke's Hospital, Cambridge, UK.
³ Genome Integrity, Immunity and Cancer Unit, Department of Immunology, Department of Genomes and Genetics, Institut Pasteur, Paris, France.
⁴ Division of Oncogenomics, The Netherlands Cancer Institute, Plesmanlaan , Amsterdam, the Netherlands.
⁵ Department of Oncology and Cancer Research UK Cambridge Institute, Li Ka Shing Centre, University of Cambridge, Cambridge, UK.
⁶ Institute of Molecular Biology (IMB), Mainz, Germany.
⁷ Department of Biochemistry, University of Cambridge, Cambridge, UK.
⁸ Wellcome Trust Sanger Institute, Hinxton, UK.
⁹ AstraZeneca, Waltham, MA, USA.
¹⁰ Vall d'Hebron Institute of Oncology, Barcelona, Spain.
¹¹ AstraZeneca, Cambridge, UK.
¹² Genome Integrity, Immunity and Cancer Unit, Department of Immunology, Department of Genomes and Genetics, Institut Pasteur, Paris, France. [email protected].
¹³ Division of Oncogenomics, The Netherlands Cancer Institute, Plesmanlaan , Amsterdam, the Netherlands. [email protected].
¹⁴ The Wellcome Trust/Cancer Research UK Gurdon Institute and Department of Biochemistry, University of Cambridge, Cambridge, UK. [email protected].
¹⁵ The Wellcome Trust/Cancer Research UK Gurdon Institute and Department of Biochemistry, University of Cambridge, Cambridge, UK. [email protected].

^# Contributed equally.

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
BRCA1 Protein / deficiency
BRCA1 Protein / genetics*
Bone Neoplasms / drug therapy*
Bone Neoplasms / genetics
Bone Neoplasms / metabolism
Bone Neoplasms / pathology
Breast Neoplasms / drug therapy*
Breast Neoplasms / genetics
Breast Neoplasms / metabolism
Breast Neoplasms / pathology
Cell Cycle Proteins
Cell Line, Tumor
Cisplatin / pharmacology
DNA Breaks, Double-Stranded
DNA End-Joining Repair*
DNA-Binding Proteins
Dose-Response Relationship, Drug
Drug Resistance, Neoplasm* / genetics
Female
HEK293 Cells
Humans
Mad2 Proteins / genetics
Mad2 Proteins / metabolism
Mice
Multiprotein Complexes
Osteosarcoma / drug therapy*
Osteosarcoma / genetics
Osteosarcoma / metabolism
Osteosarcoma / pathology
Ovarian Neoplasms / drug therapy*
Ovarian Neoplasms / genetics
Ovarian Neoplasms / metabolism
Ovarian Neoplasms / pathology
Poly(ADP-ribose) Polymerase Inhibitors / pharmacology*
Proteins / genetics
Proteins / metabolism*
Recombinational DNA Repair*
Telomere-Binding Proteins / genetics
Telomere-Binding Proteins / metabolism
Tumor Suppressor p53-Binding Protein 1 / genetics
Tumor Suppressor p53-Binding Protein 1 / metabolism
Xenograft Model Antitumor Assays

Substances

BRCA1 Protein
BRCA1 protein, human
Cell Cycle Proteins
DNA-Binding Proteins
MAD2L2 protein, human
Mad2 Proteins
Multiprotein Complexes
Poly(ADP-ribose) Polymerase Inhibitors
Proteins
Rif1 protein, human
SHLD2 protein, human
TP53BP1 protein, human
Telomere-Binding Proteins
Tumor Suppressor p53-Binding Protein 1
Cisplatin

Abstract

Publication types

MeSH terms

Substances

Grants and funding