Stimulation by Rad52 of yeast Rad51- mediated recombination

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Abstract

In Saccharomyces cerevisiae, the RAD51 and RAD52 genes are involved in recombination and in repair of damaged DNA1,2,3. The RAD51 gene is a structural and functional homologue of the recA gene4,5 and the gene product participates in strand exchange and single-stranded-DNA-dependent ATP hydrolysis by means of nucleoprotein filament formation6,7,8,9,10,11. The RAD52 gene12 is important in RAD51-mediated recombination1,2,3. Binding of this protein to Rad51 (refs 4, 13) suggests that they cooperate in recombination. Homologues of both Rad51 and Rad52 are conserved from yeast to humans14,15,16, suggesting that the mechanisms used for pairing homologous DNA molecules during recombination may be universal in eukaryotes. Here we show that Rad52 protein stimulates Rad51 reactions and that binding to Rad51 is necessary for this stimulatory effect. We conclude that this binding is crucial in recombination and that it facilitates the formation of Rad51 nucleoprotein filaments.

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Figure 1: Rad51-mediated strand exchange in the presence of Rad52.
Figure 2: Effect of the order of RPA and/or Rad52 addition on Rad51-mediated strand exchange.
Figure 3: Effects of Rad52 and RPA proteins on Rad51 ATPase activity.
Figure 4: Effect of Rad52–327 protein on Rad51 activity.

References

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Acknowledgements

We thank J. Tomizawa and H. Ogawa for encouragement and support; N. Kleckner, D. Bishop, E. Egelman, L. S. Symington and S. Gaisor for critically reading the manuscript; A. Sugino for yeast RPA protein; R. Kolodner for the overproducer strain of RPA; and D. Bishop and S. West for communicating unpublished results. This work was supported by grants from the Ministry of Education, Science, Sports and Culture of Japan and the Human Frontier Science Organization to A.S. and T.O., and a grant from National Institute of Genetics to A.S.

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Correspondence to Akira Shinohara or Tomoko Ogawa.

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