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Originally published In Press as doi:10.1074/jbc.M203494200 on June 19, 2002

J. Biol. Chem., Vol. 277, Issue 35, 31663-31672, August 30, 2002
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Rad52 Protein Associates with Replication Protein A (RPA)-Single-stranded DNA to Accelerate Rad51-mediated Displacement of RPA and Presynaptic Complex Formation*

Tomohiko Sugiyama and Stephen C. KowalczykowskiDagger

From the Sections of Microbiology and of Molecular and Cellular Biology, and Center for Genetics and Development, University of California, Davis, California 95616-8665

The Rad51 nucleoprotein filament mediates DNA strand exchange, a key step of homologous recombination. This activity is stimulated by replication protein A (RPA), but only when RPA is introduced after Rad51 nucleoprotein filament formation. In contrast, RPA inhibits Rad51 nucleoprotein complex formation by prior binding to single-stranded DNA (ssDNA), but Rad52 protein alleviates this inhibition. Here we show that Rad51 filament formation is simultaneous with displacement of RPA from ssDNA. This displacement is initiated by a rate-limiting nucleation of Rad51 protein onto ssDNA complex, followed by rapid elongation of the filament. Rad52 protein accelerates RPA displacement by Rad51 protein. This acceleration probably involves direct interactions with both Rad51 protein and RPA. Detection of a Rad52-RPA-ssDNA co-complex suggests that this co-complex is an intermediate in the displacement process.

* This work was supported by National Institutes of Health Grants AI-18987 and GM-62653 and Human Frontier Science Program Grant RG63 (to S. C. K.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Section of Microbiology, Briggs Hall, University of California, Davis, CA 95616-8665. Tel.: 530-752-5938; Fax: 530-752-5939; E-mail: sckowalczykowski@ucdavis.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.


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