Xrcc3 Is Required for Assembly of Rad51 Complexes in Vivo*
Abstract
Rad51 is a member of a family of eukaryotic proteins related to the bacterial recombinational repair protein RecA. Rad51 protein localizes to multiple subnuclear foci in Chinese hamster ovary cells. Subnuclear Rad51 foci are induced by ionizing radiation or the DNA cross-linking agent cisplatin. Formation of these foci is likely to reflect assembly of a multimeric form of Rad51 that promotes DNA repair. Formation of damage-induced Rad51 foci does not occur in the Chinese hamster ovary cell line irs1SF, which is sensitive to DNA damaging agents. The Rad51 focus formation defect of irs1SF cells is corrected by a construct that encodes the repair protein Xrcc3. Xrcc3 is a human homolog of Rad51 previously isolated by virtue of its ability to correct the radiation sensitivity of irs1SF cells. Changes in the steady state level of Rad51 protein do not account for the irs1SF defect nor do they account for the appearance of foci following DNA damage. These results suggest that Xrcc3 is required for the assembly or stabilization of a multimeric form of Rad51 during DNA repair. Cell lines defective in two different components of DNA protein kinase formed Rad51 foci in response to damage, indicating DNA protein kinase is not required for damaged-induced mobilization of Rad51.
Footnotes
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↵* This work was supported in part by a grant from the Human Frontiers Science Program (to A. S. and D. K. B.).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.
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↵§ To whom correspondence should be addressed: University of Chicago Medical Center, 5841 S. Maryland Ave. MC1105, Chicago, IL 60637. Tel.: 773-702-9211; Fax: 773-702-1968; E-mail:dbishop{at}midway.uchicago.edu.
- DSB
- double strand breaks
- CHO
- Chinese hamster ovary
- TBS
- Tris-buffered saline
- Gy
- gray.
- Received October 7, 1997.
- Revision received April 30, 1998.
- The American Society for Biochemistry and Molecular Biology, Inc.
