Synergistic actions of Rad51 and Rad52 in recombination and DNA repair

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Abstract

In the yeast Saccharomyces cerevisiae, mutations in the genes RAD51 or RAD52 result in severe defects in genetic recombination and the repair of double-strand DNA breaks. These genes, and others of the RAD52 epistasis group (RAD50, RAD54, RAD55, RAD57, RAD59, MRE11 and XRS2), were first identified by their sensitivity to X-rays1. They were subsequently shown to be required for spontaneous and induced mitotic recombination, meiotic recombination, and mating-type switching (reviewed in ref. 2). Human homologues of RAD50, RAD51, RAD52, RAD54 and MRE11 have been identified3,4,5,6. Targeted disruption of the murine RAD51 gene results in an embryonic lethal phenotype, indicating that Rad51 protein is required during cell proliferation7,8. Biochemical studies have shown that human RAD51 encodes a protein of relative molecular mass 36,966 (hRad51) that promotes ATP-dependent homologous pairing and DNA strand exchange9,10,11. As a structural and functional homologue of the RecA protein from Escherichia coli3,9,12, hRad51 is thought to play a central role in recombination. Yeast Rad51 has been shown to interact with Rad52 protein13,14,15, as does the human homologue16. Here we show that hRad52 stimulates homologous pairing by hRad51. The DNA-binding properties of hRad52 indicate that Rad52 is involved in an early stage of Rad51-mediated recombination.

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Figure 1: Purification of recombinant human Rad52 protein from E.coli.
Figure 2: Stimulation of hRad51-mediated joint molecule formation by hRad52.
Figure 3: DNA-binding properties of hRad52.
Figure 4: hRad52 is involved in the early stages of hRad51-mediated pairing reactions.

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Acknowledgements

We thank O. Bezzubova and J.-M. Buerstedde for plasmid pCR52; E. Van Dyck for contributions relating to the DNA-binding assays; and D. Bishop, A. Shinohara, T. Ogawa, P. Sung and S.Kowalczykowski for information relating to the actions of yeast Rad52 before publication. This work was supported by the ICRF and HFSPO.

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Correspondence to Stephen C. West.

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