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PMID: 21893598 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Cdk1 uncouples CtIP-dependent resection and Rad51 filament formation during M-phase double-strand break repair.

The Journal of cell biology ·Vol. 194 ·No. 5 ·2011-09-05 ·Pages 705-20

Peterson SE, Li Y, Chait BT, Gottesman ME, Baer R, Gautier J

Abstract

DNA double-strand break (DSB) resection, which results in RPA-bound single-stranded DNA (ssDNA), is activated in S phase by Cdk2. RPA-ssDNA activates the ATR-dependent checkpoint and homology-directed repair (HDR) via Rad51-dependent mechanisms. On the other hand, the fate of DSBs sustained during vertebrate M phase is largely unknown. We use cell-free Xenopus laevis egg extracts to examine the recruitment of proteins to chromatin after DSB formation. We find that S-phase extract recapitulates a two-step resection mechanism. M-phase chromosomes are also resected in cell-free extracts and cultured human cells. In contrast to the events in S phase, M-phase resection is solely dependent on MRN-CtIP. Despite generation of RPA-ssDNA, M-phase resection does not lead to ATR activation or Rad51 chromatin association. Remarkably, we find that Cdk1 permits resection by phosphorylation of CtIP but also prevents Rad51 binding to the resected ends. We have thus identified Cdk1 as a critical regulator of DSB repair in M phase. Cdk1 induces persistent ssDNA-RPA overhangs in M phase, thereby preventing both classical NHEJ and Rad51-dependent HDR.

MeSH Terms
Animals Antigens, Nuclear/metabolism Ataxia Telangiectasia Mutated Proteins CDC2 Protein Kinase/antagonists & inhibitors,metabolism Cell Cycle Proteins/metabolism Cell Division/physiology Cell Nucleus/metabolism Cell-Free System Checkpoint Kinase 1 Chromatin/metabolism Cyclin-Dependent Kinase 2/antagonists & inhibitors,metabolism DNA Breaks, Double-Stranded DNA Helicases/metabolism DNA Repair/physiology DNA, Single-Stranded/metabolism DNA-Binding Proteins/metabolism Endonucleases/metabolism HeLa Cells Histones/metabolism Humans Ku Autoantigen MRE11 Homologue Protein Meiosis/physiology Mitosis/physiology Nuclear Envelope/physiology Ovum Phosphorylation/drug effects Protein Binding/drug effects,physiology Protein Kinase Inhibitors/pharmacology Protein Kinases/metabolism Protein Serine-Threonine Kinases/metabolism Rad51 Recombinase/metabolism RecQ Helicases/metabolism Replication Protein A/metabolism S Phase/physiology Tumor Suppressor Proteins/metabolism Werner Syndrome Helicase Xenopus Proteins/antagonists & inhibitors,metabolism Xenopus laevis
Chemicals
Antigens, Nuclear Cell Cycle Proteins Chromatin DNA, Single-Stranded DNA-Binding Proteins Histones Mre11 protein, Xenopus Protein Kinase Inhibitors RBBP8 protein, Xenopus Replication Protein A Tumor Suppressor Proteins Xenopus Proteins Protein Kinases Atr protein, Xenopus ATM protein, human Ataxia Telangiectasia Mutated Proteins Checkpoint Kinase 1 Protein Serine-Threonine Kinases CDC2 Protein Kinase Cdk2 protein, Xenopus Cyclin-Dependent Kinase 2 RAD51 protein, Xenopus Rad51 Recombinase Endonucleases MRE11 Homologue Protein Bloom syndrome protein DNA Helicases DNA2 protein, Xenopus RecQ Helicases Werner Syndrome Helicase Xrcc6 protein, human Ku Autoantigen WRN protein, Xenopus
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Peterson Shaun E
Institute for Cancer Genetics, Columbia University Medical Center, New York, NY 10032, USA.
Li Yinyin
Chait Brian T
Gottesman Max E
Baer Richard
Gautier Jean
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
1540-8140
DOI
10.1083/jcb.201103103
Published
2011-09-05
Pages
705-20
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC3171114
Subset
IM
Grants
NCI NIH HHS · P01 CA097403 · United States
NIGMS NIH HHS · R01GM077495 · United States
NCI NIH HHS · R01 CA092245 · United States
NCRR NIH HHS · RR022220 · United States
NCRR NIH HHS · P41 RR000862 · United States
NCRR NIH HHS · U54 RR022220 · United States
NCRR NIH HHS · RR00862 · United States
NIGMS NIH HHS · R01 GM077495 · United States
NCI NIH HHS · R01CA092245 · United States
External Links
PubMed source DOI: 10.1083/jcb.201103103 PMC Full Text View journal details
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