CRL4(Wdr70) regulates H2B monoubiquitination and facilitates Exo1-dependent resection

Ming Zeng; Laifeng Ren; Ken'Ichi Mizuno; Konstantinos Nestoras; Haibin Wang; Zizhi Tang; Liandi Guo; Daochun Kong; Qiwen Hu; Qun He; Lilin Du; Antony M Carr; Cong Liu

doi:10.1038/ncomms11364

CRL4(Wdr70) regulates H2B monoubiquitination and facilitates Exo1-dependent resection

Nat Commun. 2016 Apr 21:7:11364. doi: 10.1038/ncomms11364.

Authors

Ming Zeng¹, Laifeng Ren², Ken'Ichi Mizuno³, Konstantinos Nestoras³, Haibin Wang¹, Zizhi Tang¹, Liandi Guo¹, Daochun Kong⁴, Qiwen Hu⁵, Qun He⁶, Lilin Du⁷, Antony M Carr³, Cong Liu¹

Affiliations

¹ Key Laboratory of Birth Defects and Related Diseases of Women and Children (Ministry of Education), Department of Paediatrics, West China Second University Hospital, Sichuan University, Chengdu 610041, China.
² Department of Medical Microbiology and Immunology, Fenyang College of Shanxi Medical University, Fenyang 032200, China.
³ Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton BN1 9RQ, UK.
⁴ National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China.
⁵ Department of Microbiology, College of Basic Medical Sciences, Third Military Medical University, Chongqing 400038, China.
⁶ State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100083, China.
⁷ National Institute of Biological Sciences, 7 Science Park Road, Zhongguancun Life Science Park, Beijing 102206, China.

Abstract

Double-strand breaks repaired by homologous recombination (HR) are first resected to form single-stranded DNA, which binds replication protein A (RPA). RPA attracts mediators that load the Rad51 filament to promote strand invasion, the defining feature of HR. How the resection machinery navigates nucleosome-packaged DNA is poorly understood. Here we report that in Schizosaccharomyces pombe a conserved DDB1-CUL4-associated factor (DCAF), Wdr70, is recruited to DSBs as part of the Cullin4-DDB1 ubiquitin ligase (CRL4(Wdr70)) and stimulates distal H2B lysine 119 mono-ubiquitination (uH2B). Wdr70 deletion, or uH2B loss, results in increased loading of the checkpoint adaptor and resection inhibitor Crb2(53BP1), decreased Exo1 association and delayed resection. Wdr70 is dispensable for resection upon Crb2(53BP1) loss, or when the Set9 methyltransferase that creates docking sites for Crb2 is deleted. Finally, we establish that this histone regulatory cascade similarly controls DSB resection in human cells.

Publication types

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

MeSH terms

Amino Acid Sequence
Cell Cycle Proteins / genetics
Cell Cycle Proteins / metabolism
DNA / genetics*
DNA / metabolism
DNA Breaks, Double-Stranded
DNA, Single-Stranded / genetics
DNA, Single-Stranded / metabolism
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism
Exodeoxyribonucleases / genetics*
Exodeoxyribonucleases / metabolism
HEK293 Cells
Histone-Lysine N-Methyltransferase / genetics
Histone-Lysine N-Methyltransferase / metabolism
Humans
Molecular Sequence Data
Nuclear Proteins / genetics
Nuclear Proteins / metabolism
Rad51 Recombinase / genetics
Rad51 Recombinase / metabolism
Recombinational DNA Repair*
Replication Protein A / genetics
Replication Protein A / metabolism
Schizosaccharomyces / genetics*
Schizosaccharomyces / metabolism
Schizosaccharomyces pombe Proteins / genetics
Schizosaccharomyces pombe Proteins / metabolism
Sequence Alignment
Signal Transduction
Ubiquitin-Protein Ligases / deficiency
Ubiquitin-Protein Ligases / genetics*
Ubiquitination

Substances

Cell Cycle Proteins
Crb2 protein, S pombe
DNA, Single-Stranded
DNA-Binding Proteins
Ddb1 protein, S pombe
Nuclear Proteins
RHP51 protein, S pombe
Replication Protein A
Schizosaccharomyces pombe Proteins
DNA
Histone-Lysine N-Methyltransferase
Ubiquitin-Protein Ligases
Rad51 Recombinase
Exodeoxyribonucleases
exodeoxyribonuclease I

Abstract

Publication types

MeSH terms

Substances

Grants and funding