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PMID: 18337252 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Rad51 protein controls Rad52-mediated DNA annealing.

The Journal of biological chemistry ·Vol. 283 ·No. 21 ·2008-05-23 ·Pages 14883-92

Wu Y, Kantake N, Sugiyama T, Kowalczykowski SC

Abstract

In Saccharomyces cerevisiae, Rad52 protein plays an essential role in the repair of DNA double-stranded breaks (DSBs). Rad52 and its orthologs possess the unique capacity to anneal single-stranded DNA (ssDNA) complexed with its cognate ssDNA-binding protein, RPA. This annealing activity is used in multiple mechanisms of DSB repair: single-stranded annealing, synthesis-dependent strand annealing, and cross-over formation. Here we report that the S. cerevisiae DNA strand exchange protein, Rad51, prevents Rad52-mediated annealing of complementary ssDNA. Efficient inhibition is ATP-dependent and involves a specific interaction between Rad51 and Rad52. Free Rad51 can limit DNA annealing by Rad52, but the Rad51 nucleoprotein filament is even more effective. We also discovered that the budding yeast Rad52 paralog, Rad59 protein, partially restores Rad52-dependent DNA annealing in the presence of Rad51, suggesting that Rad52 and Rad59 function coordinately to enhance recombinational DNA repair either by directing the processed DSBs to repair by DNA strand annealing or by promoting second end capture to form a double Holliday junction. This regulation of Rad52-mediated annealing suggests a control function for Rad51 in deciding the recombination path taken for a processed DNA break; the ssDNA can be directed to either Rad51-mediated DNA strand invasion or to Rad52-mediated DNA annealing. This channeling determines the nature of the subsequent repair process and is consistent with the observed competition between these pathways in vivo.

MeSH Terms
Base Pairing Cell Nucleus/metabolism DNA, Single-Stranded/chemistry,metabolism DNA-Binding Proteins/genetics,metabolism Protein Binding Rad51 Recombinase/metabolism Rad52 DNA Repair and Recombination Protein/antagonists & inhibitors,metabolism Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins/genetics,metabolism
Chemicals
DNA, Single-Stranded DNA-Binding Proteins RAD59 protein, S cerevisiae Rad52 DNA Repair and Recombination Protein Saccharomyces cerevisiae Proteins Rad51 Recombinase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wu Yun
Sections of Microbiology and of Molecular and Cellular Biology, University of California, Davis, CA 95616-8665, USA.
Kantake Noriko
Sugiyama Tomohiko
Kowalczykowski Stephen C
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
DOI
10.1074/jbc.M801097200
Published
2008-05-23
Epub
2008-00-12
Pages
14883-92
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2386943
Subset
IM
Grants
NIAID NIH HHS · AI 18987 · United States
External Links
PubMed source DOI: 10.1074/jbc.M801097200 PMC Full Text View journal details
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