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Human meiotic recombinase Dmc1 promotes ATP-dependent homologous DNA strand exchange

Nature volume 429, pages 433–437 (2004)Cite this article

Abstract

Homologous recombination is crucial for the repair of DNA breaks and maintenance of genome stability1,2,3. In Escherichia coli, homologous recombination is dependent on the RecA protein. In the presence of ATP, RecA mediates the homologous DNA pairing and strand exchange reaction that links recombining DNA molecules. DNA joint formation is initiated through the nucleation of RecA onto single-stranded DNA (ssDNA) to form helical nucleoprotein filaments4,5,6. Two RecA-like recombinases, Rad51 and Dmc1, exist in eukaryotes1. Whereas Rad51 is needed for both mitotic and meiotic recombination events, the function of Dmc1 is restricted to meiosis3,7. Here we examine human Dmc1 protein (hDmc1) for the ability to promote DNA strand exchange, and show that hDmc1 mediates strand exchange between paired DNA substrates over at least several thousand base pairs. DNA strand exchange requires ATP and is strongly dependent on the heterotrimeric ssDNA-binding molecule replication factor A (RPA). We present evidence that hDmc1-mediated DNA recombination initiates through the nucleation of hDmc1 onto ssDNA to form a helical nucleoprotein filament. The DNA strand exchange activity of hDmc1 is probably indispensable for repair of DNA double-strand breaks during meiosis and for maintaining the ploidy of meiotic chromosomes.

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Figure 1: hDmc1 catalyses ATP-dependent DNA strand exchange.
Figure 2: Dependence of hDmc1-mediated DNA strand exchange on hRPA and salt.
Figure 3: hDmc1 mediates robust D-loop formation.
Figure 4: Analysis of hDmc1-ssDNA complexes by electron microscopy.

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References

  1. Masson, J. Y. & West, S. C. The Rad51 and Dmc1 recombinases: a non-identical twin relationship. Trends Biochem. Sci. 26, 131–136 (2001)

    Article  CAS  Google Scholar 

  2. Sung, P., Krejci, L., Van Komen, S. & Sehorn, M. G. Rad51 recombinase and recombination mediators. J. Biol. Chem. 278, 42729–42732 (2003)

    Article  CAS  Google Scholar 

  3. Symington, L. S. Role of RAD52 epistasis group genes in homologous recombination and double-strand break repair. Microbiol. Mol. Biol. Rev. 66, 630–670 (2002)

    Article  CAS  Google Scholar 

  4. Bianco, P. R., Tracy, R. B. & Kowalczykowski, S. C. DNA strand exchange proteins: a biochemical and physical comparison. Front. Biosci. 3, D570–D603 (1998)

    Article  CAS  Google Scholar 

  5. Roca, A. I. & Cox, M. M. RecA protein: structure, function, and role in recombinational DNA repair. Prog. Nucleic Acid Res. Mol. Biol. 56, 129–223 (1997)

    Article  CAS  Google Scholar 

  6. Yu, X., Jacobs, S. A., West, S. C., Ogawa, T. & Egelman, E. H. Domain structure and dynamics in the helical filaments formed by RecA and Rad51 on DNA. Proc. Natl Acad. Sci. USA 98, 8419–8424 (2001)

    Article  ADS  CAS  Google Scholar 

  7. Bishop, D. K., Park, D., Xu, L. & Kleckner, N. DMC1: a meiosis-specific yeast homolog of E. coli recA required for recombination, synaptonemal complex formation, and cell cycle progression. Cell 69, 439–456 (1992)

    Article  CAS  Google Scholar 

  8. Shinohara, A. et al. Cloning of human, mouse and fission yeast recombination genes homologous to RAD51 and recA. Nature Genet. 4, 239–243 (1993)

    Article  CAS  Google Scholar 

  9. Lydall, D., Nikolsky, Y., Bishop, D. K. & Weinert, T. A meiotic recombination checkpoint controlled by mitotic checkpoint genes. Nature 383, 840–843 (1996)

    Article  ADS  CAS  Google Scholar 

  10. Yoshida, K. et al. The mouse RecA-like gene Dmc1 is required for homologous chromosome synapsis during meiosis. Mol. Cell 1, 707–718 (1998)

    Article  CAS  Google Scholar 

  11. Pittman, D. L. et al. Meiotic prophase arrest with failure of chromosome synapsis in mice deficient for Dmc1, a germline-specific RecA homolog. Mol. Cell 1, 697–705 (1998)

    Article  CAS  Google Scholar 

  12. Li, Z., Golub, E. I., Gupta, R. & Radding, C. M. Recombination activities of HsDmc1 protein, the meiotic human homolog of RecA protein. Proc. Natl Acad. Sci. USA 94, 11221–11226 (1997)

    Article  ADS  CAS  Google Scholar 

  13. Masson, J. Y. et al. The meiosis-specific recombinase hDmc1 forms ring structures and interacts with hRad51. EMBO J. 18, 6552–6560 (1999)

    Article  CAS  Google Scholar 

  14. Hong, E. L., Shinohara, A. & Bishop, D. K. Saccharomyces cerevisiae Dmc1 protein promotes renaturation of single-strand DNA (ssDNA) and assimilation of ssDNA into homologous super-coiled duplex DNA. J. Biol. Chem. 276, 41906–41912 (2001)

    Article  CAS  Google Scholar 

  15. Shinohara, A., Ogawa, H. & Ogawa, T. Rad51 protein involved in repair and recombination in S. cerevisiae is a RecA-like protein. Cell 69, 457–470 (1992)

    Article  CAS  Google Scholar 

  16. Passy, S. I. et al. Human Dmc1 protein binds DNA as an octameric ring. Proc. Natl Acad. Sci. USA 96, 10684–10688 (1999)

    Article  ADS  CAS  Google Scholar 

  17. Gupta, R. C., Golub, E., Bi, B. & Radding, C. M. The synaptic activity of HsDmc1, a human recombination protein specific to meiosis. Proc. Natl Acad. Sci. USA 98, 8433–8439 (2001)

    Article  ADS  CAS  Google Scholar 

  18. Sigurdsson, S., Trujillo, K., Song, B., Stratton, S. & Sung, P. Basis for avid homologous DNA strand exchange by human Rad51 and RPA. J. Biol. Chem. 276, 8798–8806 (2001)

    Article  CAS  Google Scholar 

  19. Van Komen, S., Petukhova, G., Sigurdsson, S. & Sung, P. Functional crosstalk among Rad51, Rad54, and RPA in heteroduplex DNA joint formation. J. Biol. Chem. 277, 43578–43587 (2002)

    Article  CAS  Google Scholar 

  20. Eggler, A. L., Inman, R. B. & Cox, M. M. The Rad51-dependent pairing of long DNA substrates is stabilized by replication protein A. J. Biol. Chem. 277, 39280–39288 (2002)

    Article  CAS  Google Scholar 

  21. Zickler, D. & Kleckner, N. Meiotic chromosomes: integrating structure and function. Annu. Rev. Genet. 33, 603–754 (1999)

    Article  CAS  Google Scholar 

  22. Henricksen, L. A., Umbricht, C. B. & Wold, M. S. Recombinant replication protein A: expression, complex formation, and functional characterization. J. Biol. Chem. 269, 11121–11132 (1994)

    CAS  PubMed  Google Scholar 

  23. Sigurdsson, S., Van Komen, S., Petukhova, G. & Sung, P. Homologous DNA pairing by human recombination factors Rad51 and Rad54. J. Biol. Chem. 277, 42790–42794 (2002)

    Article  CAS  Google Scholar 

  24. Sung, P. & Robberson, D. L. DNA strand exchange mediated by a RAD51-ssDNA nucleoprotein filament with polarity opposite to that of RecA. Cell 82, 453–461 (1995)

    Article  CAS  Google Scholar 

  25. Crowther, R. A., Henderson, R. & Smith, J. M. MRC image processing programs. J. Struct. Biol. 116, 9–16 (1996)

    Article  CAS  Google Scholar 

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Acknowledgements

We thank T. Habu for helping with cDNA cloning and baculovirus construction. This work was supported by grants from the US National Institutes of Health (P.S. and V.M.U.).

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Author notes

  1. Michael G. Sehorn and Stefan Sigurdsson: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, 333 Cedar Street, New Haven, Connecticut, 06520, USA

    Michael G. Sehorn, Vinzenz M. Unger & Patrick Sung

  2. Department of Molecular Medicine, Institute of Biotechnology, University of Texas Health Science Center at San Antonio, 15355 Lambda Drive, San Antonio, Texas, 78245, USA

    Stefan Sigurdsson & Wendy Bussen

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  1. Michael G. Sehorn
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Corresponding author

Correspondence to Patrick Sung.

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The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Figure 1

DNA strand exchange reactions that used the 3' and 5' end-labeled duplex molecules. (JPG 30 kb)

Supplementary Figure 2

This figure shows a physical and functional interaction of Rad54B with hDmc1. (JPG 43 kb)

Supplementary Information

Methods and legends for Supplementary Figures 1 and 2. (DOC 33 kb)

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Sehorn, M., Sigurdsson, S., Bussen, W. et al. Human meiotic recombinase Dmc1 promotes ATP-dependent homologous DNA strand exchange. Nature 429, 433–437 (2004). https://doi.org/10.1038/nature02563

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