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Originally published In Press as doi:10.1074/jbc.M002075200 on June 27, 2000

J. Biol. Chem., Vol. 275, Issue 37, 29100-29106, September 15, 2000
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The RAD51 Family Member, RAD51L3, Is a DNA-stimulated ATPase That Forms a Complex with XRCC2*

Jeremy P. BraybrookeDagger , Karen G. Spink§, John Thacker§, and Ian D. HicksonDagger

From the Dagger  Imperial Cancer Research Fund Laboratories, Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS and § Medical Research Council Radiation and Genome Stability Unit, Harwell, Oxfordshire OX11 ORD, United Kingdom

The Rad51 protein in eukaryotic cells is a structural and functional homolog of Escherichia coli RecA with a role in DNA repair and genetic recombination. Several proteins showing sequence similarity to Rad51 have previously been identified in both yeast and human cells. In Saccharomyces cerevisiae, two of these proteins, Rad55p and Rad57p, form a heterodimer that can stimulate Rad51-mediated DNA strand exchange. Here, we report the purification of one of the representatives of the RAD51 family in human cells. We demonstrate that the purified RAD51L3 protein possesses single-stranded DNA binding activity and DNA-stimulated ATPase activity, consistent with the presence of "Walker box" motifs in the deduced RAD51L3 sequence. We have identified a protein complex in human cells containing RAD51L3 and a second RAD51 family member, XRCC2. By using purified proteins, we demonstrate that the interaction between RAD51L3 and XRCC2 is direct. Given the requirements for XRCC2 in genetic recombination and protection against DNA-damaging agents, we suggest that the complex of RAD51L3 and XRCC2 is likely to be important for these functions in human cells.

* This work was supported by the Imperial Cancer Research Fund (to J. P. B. and I. D. H.), the Medical Research Council, and the European Commission (to K. G. S. and J. T.).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.

To whom correspondence should be addressed. Tel.: 44-1865-222417; Fax: 44-1865-222431; E-mail: hickson@icrf.icnet.uk.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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