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Originally published In Press as doi:10.1074/jbc.M205864200 on September 10, 2002

J. Biol. Chem., Vol. 277, Issue 46, 43578-43587, November 15, 2002
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Functional Cross-talk among Rad51, Rad54, and Replication Protein A in Heteroduplex DNA Joint Formation*

Stephen Van KomenDagger , Galina Petukhova§, Stefan Sigurdsson, and Patrick Sung||

From the Department of Molecular Medicine/Institute of Biotechnology, University of Texas Health Science Center at San Antonio, San Antonio, Texas 78245-3207

Saccharomyces cerevisiae Rad51, Rad54, and replication protein A (RPA) proteins work in concert to make heteroduplex DNA joints during homologous recombination. With plasmid length DNA substrates, maximal DNA joint formation is observed with amounts of Rad51 substantially below what is needed to saturate the initiating single-stranded DNA template, and, relative to Rad51, Rad54 is needed in only catalytic quantities. RPA is still indispensable for optimal reaction efficiency, but its role in this instance is to sequester free single-stranded DNA, which otherwise inhibits Rad51 and Rad54 functions. We also demonstrate that Rad54 helps overcome various reaction constraints in DNA joint formation. These results thus shed light on the function of Rad54 in the Rad51-mediated homologous DNA pairing reaction and also reveal a novel role of RPA in the presynaptic stage of this reaction.

* This work was supported by United States Public Health Service Grants RO1ES07061 and RO1GM57814.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 Supported in part by United States Army Predoctoral Fellowship DAMD17-01-1-0414.

§ Present Address: NIDDK, National Institutes of Health, Bldg. 10, Rm. 9D17, 9000 Rockville Pike, Bethesda, MD 20892.

Supported in part by United States Army Training Grant DAMD17-99-1-9402 and Predoctoral Fellowship DAMD17-01-1-0412.

|| To whom correspondence should be addressed: Dept. of Molecular Medicine/Institute of Biotechnology, University of Texas Health Science Center at San Antonio, 15355 Lambda Dr., San Antonio, TX 78245-3207. Tel.: 210-567-7216; Fax: 210-567-7277; E-mail: sung@uthscsa.edu.

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


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