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J. Biol. Chem., Vol. 282, Issue 3, 1973-1979, January 19, 2007

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Role of RAD51C and XRCC3 in Genetic Recombination and DNA Repair^*

From the Clare Hall Laboratories, London Research Institute, Cancer Research UK, South Mimms, Hertfordshire EN6 3LD, United Kingdom

In germ line cells, recombination is required for gene reassortment and proper chromosome segregation at meiosis, whereas in somatic cells it provides an important mechanism for the repair of DNA double-strand breaks. Five proteins (RAD51B, RAD51C, RAD51D, XRCC2, and XRCC3) that share homology with RAD51 recombinase and are known as the RAD51 paralogs are important for recombinational repair, as paralog-defective cell lines exhibit spontaneous chromosomal aberrations, defective DNA repair, and reduced gene targeting. The paralogs form two distinct protein complexes, RAD51B-RAD51C-RAD51D-XRCC2 and RAD51C-XRCC3, but their precise cellular roles remain unknown. Here, we show that, like MLH1, RAD51C localized to mouse meiotic chromosomes at pachytene/diplotene. Using immunoprecipitation and gel filtration analyses, we found that Holliday junction resolvase activity associated tightly and co-eluted with the 80-kDa RAD51C-XRCC3 complex. Taken together, these data indicate that the RAD51C-XRCC3-associated Holliday junction resolvase complex associates with crossovers and may play an essential role in the resolution of recombination intermediates prior to chromosome segregation.

Received for publication, September 25, 2006 , and in revised form, October 27, 2006.

^* This work was supported in part by Cancer Research UK, the European Union DNA Repair Consortium, and the Breast Cancer Campaign. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement"in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Both authors contributed equally to this work.

² Supported in part by a fellowship from the American Cancer Society. Present address: Dept. of Therapeutic Radiology, Yale University School of Medicine, New Haven, CT 06510.

³ Present address: Dept. of Radiation Oncology and Biology, John Radcliffe Hospital, University of Oxford, Oxfordshire OX3 9DU, UK.

⁴ Present address: Pfizer, Sandwich, Kent CT13 9NJ, UK.

⁵ To whom correspondence should be addressed. Tel.: 44-1707-625868; Fax: 44-1707-625811; E-mail: stephen.west{at}cancer.org.uk.

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