Article
- The EMBO Journal (2006) 25, 5539 - 5548
- doi:10.1038/sj.emboj.7601412
Published online: 9 November 2006
Subject Category:
Rad52-mediated DNA annealing after Rad51-mediated DNA strand exchange promotes second ssDNA capture
Tomohiko Sugiyama1, Noriko Kantake1, Yun Wu2,3,a and Stephen C Kowalczykowski2
- Department of Biological Sciences, Ohio University, Athens, OH, USA
- Sections of Microbiology, and Molecular and Cellular Biology, Center for Genetics and Development, University of California, Davis, CA, USA
- Microbiology Graduate group, University of California, Davis, CA, USA
Correspondence to:
Tomohiko Sugiyama, Department of Biological Sciences, Ohio University, 211 Life Science Research Facility, Athens, OH 45701, USA. Tel.: +1 740 597 1927; Fax: +1 740 593 0300; E-mail: sugiyama@ohio.edu
aPresent address: Department of Molecular Biology, Lewis Thomas Labs, Princeton University, Princeton, NJ 08544, USA
Received 17 May 2006; Accepted 10 October 2006
Abstract
Rad51, Rad52, and RPA play central roles in homologous DNA recombination. Rad51 mediates DNA strand exchange, a key reaction in DNA recombination. Rad52 has two distinct activities: to recruit Rad51 onto single-strand (ss)DNA that is complexed with the ssDNA-binding protein, RPA, and to anneal complementary ssDNA complexed with RPA. Here, we report that Rad52 promotes annealing of the ssDNA strand that is displaced by DNA strand exchange by Rad51 and RPA, to a second ssDNA strand. An RPA that is recombination-deficient (RPA(rfa1-t11)) failed to support annealing, explaining its in vivo phenotype. Escherichia coli RecO and SSB proteins, which are functional homologues of Rad52 and RPA, also facilitated the same reaction, demonstrating its conserved nature. We also demonstrate that the two activities of Rad52, recruiting Rad51 and annealing DNA, are coordinated in DNA strand exchange and second ssDNA capture.
Keywords:
- DNA annealing,
- DNA repair,
- Rad52,
- recombination,
- second ssDNA capture
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