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The Srs2 helicase prevents recombination by disrupting Rad51 nucleoprotein filaments

Abstract

Homologous recombination is a ubiquitous process with key functions in meiotic and vegetative cells for the repair of DNA breaks. It is initiated by the formation of single-stranded DNA on which recombination proteins bind to form a nucleoprotein filament that is active in searching for homology, in the formation of joint molecules and in the exchange of DNA strands1. This process contributes to genome stability but it is also potentially dangerous to cells if intermediates are formed that cannot be processed normally and thus are toxic or generate genomic rearrangements. Cells must therefore have developed strategies to survey recombination and to prevent the occurrence of such deleterious events. In Saccharomyces cerevisiae, genetic data have shown that the Srs2 helicase negatively modulates recombination2,3, and later experiments suggested that it reverses intermediate recombination structures4,5,6,7. Here we show that DNA strand exchange mediated in vitro by Rad51 is inhibited by Srs2, and that Srs2 disrupts Rad51 filaments formed on single-stranded DNA. These data provide an explanation for the anti-recombinogenic role of Srs2 in vivo and highlight a previously unknown mechanism for recombination control.

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Figure 1: Purification, ATPase and helicase activities of Srs2.
Figure 2: Srs2 inhibits DNA strand exchange catalysed by Rad51.
Figure 3: Disruption of Rad51 presynaptic filament by Srs2 as examined by electron microscopy.

References

  1. Sung, P., Trujillo, K. M. & Van Komen, S. Recombination factors of Saccharomyces cerevisiae. Mutat. Res. 451, 257–275 (2000)

    CAS  Article  Google Scholar 

  2. Aguilera, A. & Klein, H. L. Genetic control of intrachromosomal recombination in Saccharomyces cerevisiae. I. Isolation and genetic characterization of hyper-recombination mutations. Genetics 119, 779–790 (1988)

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Aboussekhra, A. et al. RADH, a gene of Saccharomyces cerevisiae encoding a putative DNA helicase involved in DNA repair. Characteristics of radH mutants and sequence of the gene. Nucleic Acids Res. 17, 7211–7219 (1989)

    CAS  Article  Google Scholar 

  4. Kaytor, M. D., Nguyen, M. & Livingston, D. M. The complexity of the interaction between RAD52 and SRS2. Genetics 140, 1441–1442 (1995)

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Milne, G. T., Ho, T. & Weaver, D. T. Modulation of Saccharomyces cerevisiae DNA double-strand break repair by SRS2 and RAD51. Genetics 139, 1189–1199 (1995)

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Schild, D. Suppression of a new allele of the yeast RAD52 gene by overexpression of RAD51, mutations in srs2 and ccr4, or mating-type heterozygosity. Genetics 140, 115–127 (1995)

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Chanet, R., Heude, M., Adjiri, A., Maloisel, L. & Fabre, F. Semidominant mutations in the yeast Rad51 protein and their relationships with the Srs2 helicase. Mol. Cell. Biol. 16, 4782–4789 (1996)

    CAS  Article  Google Scholar 

  8. Fabre, F., Chan, A., Heyer, W. D. & Gangloff, S. Alternate pathways involving Sgs1/Top3, Mus81/Mms4, and Srs2 prevent formation of toxic recombination intermediates from single-stranded gaps created by DNA replication. Proc. Natl Acad. Sci. USA 99, 1687–1692 (2002)

    Article  Google Scholar 

  9. Klein, H. L. Mutations in recombinational repair and in checkpoint control genes suppress the lethal combination of srs2Δ with other DNA repair genes in Saccharomyces cerevisiae. Genetics 157, 557–565 (2001)

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Singleton, M. R. & Wigley, D. B. Modularity and specialization in superfamily 1 and 2 helicases. J. Bacteriol. 184, 1819–1826 (2002)

    CAS  Article  Google Scholar 

  11. Rong, L. & Klein, H. L. Purification and characterization of the Srs2 DNA helicase of the yeast Saccharomyces cerevisiae. J. Biol. Chem. 268, 1252–1259 (1993)

    CAS  PubMed  Google Scholar 

  12. Petukhova, G., Stratton, S. & Sung, P. Catalysis of homologous DNA pairing by yeast Rad51 and Rad54 proteins. Nature 393, 91–94 (1998)

    ADS  CAS  Article  Google Scholar 

  13. Krejci, L. et al. DNA helicase Srs2 disrupts formation of the Rad51 presynaptic filament. Nature 423, 305–309 (2003)

    ADS  CAS  Article  Google Scholar 

  14. Aboussekhra, A., Chanet, R., Adjiri, A. & Fabre, F. Semidominant suppressors of Srs2 helicase mutations of Saccharomyces cerevisiae map in the RAD51 gene, whose sequence predicts a protein with similarities to procaryotic RecA proteins. Mol. Cell. Biol. 12, 3224–3234 (1992)

    CAS  Article  Google Scholar 

  15. Lovett, S. T. & Mortimer, R. K. Characterization of null mutants of the RAD55 gene of Saccharomyces cerevisiae: effects of temperature, osmotic strength and mating type. Genetics 116, 547–553 (1987)

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Vaze, M. et al. Recovery from checkpoint-mediated arrest after repair of a double-strand break requires Srs2 helicase. Mol. Cell 10, 373 (2002)

    CAS  Article  Google Scholar 

  17. Palladino, F. & Klein, H. L. Analysis of mitotic and meiotic defects in Saccharomyces cerevisiae SRS2 DNA helicase mutants. Genetics 132, 23–37 (1992)

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Heude, M., Chanet, R. & Fabre, F. Regulation of the Saccharomyces cerevisiae Srs2 helicase during the mitotic cell cycle, meiosis and after irradiation. Mol. Gen. Genet. 248, 59–68 (1995)

    CAS  Article  Google Scholar 

  19. Van Komen, S., Petukhova, G., Sigurdsson, S., Stratton, S. & Sung, P. Superhelicity-driven homologous DNA pairing by yeast recombination factors Rad51 and Rad54. Mol. Cell 6, 563–572 (2000)

    CAS  Article  Google Scholar 

  20. Gangloff, S., Soustelle, C. & Fabre, F. Homologous recombination is responsible for cell death in the absence of the Sgs1 and Srs2 helicases. Nature Genet. 25, 192–194 (2000)

    CAS  Article  Google Scholar 

  21. van Brabant, A. J., Stan, R. & Ellis, N. A. DNA helicases, genomic instability, and human genetic disease. Annu. Rev. Genomics Hum. Genet. 1, 409–459 (2000)

    CAS  Article  Google Scholar 

  22. Saintigny, Y., Makienko, K., Swanson, C., Emond, M. J. & Monnat, R. J. Jr Homologous recombination resolution defect in Werner syndrome. Mol. Cell. Biol. 22, 6971–6978 (2002)

    CAS  Article  Google Scholar 

  23. Iggo, R. D. & Lane, D. P. Nuclear protein p68 is an RNA-dependent ATPase. EMBO J. 8, 1827–1831 (1989)

    CAS  Article  Google Scholar 

  24. Beloin, C. et al. Contribution of DNA conformation and topology in right-handed DNA wrapping by the Bacillus subtilis LrpC protein. J. Biol. Chem. 278, 5333–5342 (2003)

    CAS  Article  Google Scholar 

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Acknowledgements

We thank L. Miccoli, D. Biard and J. Angulo for advice on recombinant baculovirus preparation and protein purification. P. Bertrand and C. Auvin for confirming the identity of Srs2 by MALDI–TOF mass spectrometry, and S. Gangloff, N. Kantake, L. Leloup, L. Maloisel, J. New and T. Robert for comments and discussions. This work was supported by the Commissariat à l'Energie Atomique, the Centre National de la Recherche Scientifique and Électricité de France.

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Correspondence to Xavier Veaute or Francis Fabre.

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Veaute, X., Jeusset, J., Soustelle, C. et al. The Srs2 helicase prevents recombination by disrupting Rad51 nucleoprotein filaments. Nature 423, 309–312 (2003). https://doi.org/10.1038/nature01585

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