Abstract
Homologous recombination is a high-fidelity DNA repair pathway. Besides a critical role in accurate chromosome segregation during meiosis, recombination functions in DNA repair and in the recovery of stalled or broken replication forks to ensure genomic stability. In contrast, inappropriate recombination contributes to genomic instability, leading to loss of heterozygosity, chromosome rearrangements and cell death. The RecA/UvsX/RadA/Rad51 family of proteins catalyses the signature reactions of recombination, homology search and DNA strand invasion1,2. Eukaryotes also possess Rad51 paralogues, whose exact role in recombination remains to be defined3. Here we show that the Saccharomyces cerevisiae Rad51 paralogues, the Rad55–Rad57 heterodimer, counteract the antirecombination activity of the Srs2 helicase. The Rad55–Rad57 heterodimer associates with the Rad51–single-stranded DNA filament, rendering it more stable than a nucleoprotein filament containing Rad51 alone. The Rad51–Rad55–Rad57 co-filament resists disruption by the Srs2 antirecombinase by blocking Srs2 translocation, involving a direct protein interaction between Rad55–Rad57 and Srs2. Our results demonstrate an unexpected role of the Rad51 paralogues in stabilizing the Rad51 filament against a biologically important antagonist, the Srs2 antirecombination helicase. The biological significance of this mechanism is indicated by a complete suppression of the ionizing radiation sensitivity of rad55 or rad57 mutants by concomitant deletion of SRS2, as expected for biological antagonists. We propose that the Rad51 presynaptic filament is a meta-stable reversible intermediate, whose assembly and disassembly is governed by the balance between Rad55–Rad57 and Srs2, providing a key regulatory mechanism controlling the initiation of homologous recombination. These data provide a paradigm for the potential function of the human RAD51 paralogues, which are known to be involved in cancer predisposition and human disease.
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Acknowledgements
We thank M. Alexeeva for the cell culture support. We thank P. Sung, R. Kolodner and L. Symington for plasmids and yeast strains. We are grateful to S. Kowalczykowski, N. Hunter, D. Castaño-Diez, P. Ringler and all members of the Heyer laboratory for discussions and comments on the manuscript. This work was supported by Postdoctoral Fellowship 17FT-0046 from the Tobacco-Related Disease Research Program (J.L.), by the European Community (LSHG-CT-2003-503303) and the Centre National de la Recherche Scientifique, the Commissariat à l'Energie Atomique (X.V., F.F.), by SystemsX.ch (H.S.), and the National Institutes of Health grants U54GM74929 (H.S.), CA92267 and GM58015 (W.-D.H.).
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J.L. designed, performed and analysed all experiments, except the ionizing radiation survival assay, and helped write the manuscript. L.R. helped with the electron microscopy image collection and data analysis. X.V. purified the Srs2 protein. F.F. performed the ionizing radiation experiment. H.S. advised on the electron microscopy analysis. W.-D.H. conceived the project, designed experiments, coordinated collaborations, contributed to data analysis and wrote the manuscript. All authors discussed results and edited the manuscript.
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Liu, J., Renault, L., Veaute, X. et al. Rad51 paralogues Rad55–Rad57 balance the antirecombinase Srs2 in Rad51 filament formation. Nature 479, 245–248 (2011). https://doi.org/10.1038/nature10522
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DOI: https://doi.org/10.1038/nature10522
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