CRL4(Wdr70) regulates H2B monoubiquitination and facilitates Exo1-dependent resection.
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.
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.
Ming
M
Zeng
Department of Medical Microbiology and Immunology, Fenyang College of Shanxi Medical University, Fenyang 032200, China.
Laifeng
L
Ren
Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton BN1 9RQ, UK.
Ken'Ichi
K
Mizuno
Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton BN1 9RQ, UK.
Konstantinos
K
Nestoras
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.
Haibin
H
Wang
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.
Zizhi
Z
Tang
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.
Liandi
L
Guo
National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China.
Daochun
D
Kong
Department of Microbiology, College of Basic Medical Sciences, Third Military Medical University, Chongqing 400038, China.
Qiwen
Q
Hu
State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100083, China.
Qun
Q
He
National Institute of Biological Sciences, 7 Science Park Road, Zhongguancun Life Science Park, Beijing 102206, China.
Lilin
L
Du
Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton BN1 9RQ, UK.
Antony M
AM
Carr
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.
Cong
C
Liu
D000595:Amino Acid Sequence; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded; D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005090:Exodeoxyribonucleases / Q000235:genetics* / Q000378:metabolism; D057809:HEK293 Cells; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D051135:Rad51 Recombinase / Q000235:genetics / Q000378:metabolism; D059767:Recombinational DNA Repair; D051716:Replication Protein A / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D015398:Signal Transduction; D044767:Ubiquitin-Protein Ligases / Q000172:deficiency / Q000235:genetics*; D054875:Ubiquitination
D018797:Cell Cycle Proteins; C109867:Crb2 protein, S pombe; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; C500125:Ddb1 protein, S pombe; D009687:Nuclear Proteins; C084232:RHP51 protein, S pombe; D051716:Replication Protein A; D029702:Schizosaccharomyces pombe Proteins; D004247:DNA; D011495:Histone-Lysine N-Methyltransferase; D044767:Ubiquitin-Protein Ligases; D051135:Rad51 Recombinase; D005090:Exodeoxyribonucleases; C028450:exodeoxyribonuclease I