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.
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Masson, J. Y. & West, S. C. The Rad51 and Dmc1 recombinases: a non-identical twin relationship. Trends Biochem. Sci. 26, 131–136 (2001)
Sung, P., Krejci, L., Van Komen, S. & Sehorn, M. G. Rad51 recombinase and recombination mediators. J. Biol. Chem. 278, 42729–42732 (2003)
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)
Bianco, P. R., Tracy, R. B. & Kowalczykowski, S. C. DNA strand exchange proteins: a biochemical and physical comparison. Front. Biosci. 3, D570–D603 (1998)
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)
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)
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)
Shinohara, A. et al. Cloning of human, mouse and fission yeast recombination genes homologous to RAD51 and recA. Nature Genet. 4, 239–243 (1993)
Lydall, D., Nikolsky, Y., Bishop, D. K. & Weinert, T. A meiotic recombination checkpoint controlled by mitotic checkpoint genes. Nature 383, 840–843 (1996)
Yoshida, K. et al. The mouse RecA-like gene Dmc1 is required for homologous chromosome synapsis during meiosis. Mol. Cell 1, 707–718 (1998)
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)
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)
Masson, J. Y. et al. The meiosis-specific recombinase hDmc1 forms ring structures and interacts with hRad51. EMBO J. 18, 6552–6560 (1999)
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)
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)
Passy, S. I. et al. Human Dmc1 protein binds DNA as an octameric ring. Proc. Natl Acad. Sci. USA 96, 10684–10688 (1999)
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)
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)
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)
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)
Zickler, D. & Kleckner, N. Meiotic chromosomes: integrating structure and function. Annu. Rev. Genet. 33, 603–754 (1999)
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)
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)
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)
Crowther, R. A., Henderson, R. & Smith, J. M. MRC image processing programs. J. Struct. Biol. 116, 9–16 (1996)
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.).
The authors declare that they have no competing financial interests.
DNA strand exchange reactions that used the 3' and 5' end-labeled duplex molecules. (JPG 30 kb)
This figure shows a physical and functional interaction of Rad54B with hDmc1. (JPG 43 kb)
Methods and legends for Supplementary Figures 1 and 2. (DOC 33 kb)
About this article
Cite this article
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
The BCL-2 pathway preserves mammalian genome integrity by eliminating recombination-defective oocytes
Nature Communications (2020)
E2f5 is a versatile transcriptional activator required for spermatogenesis and multiciliated cell differentiation in zebrafish
PLOS Genetics (2020)
Proceedings of the National Academy of Sciences (2020)
Alzheimer's & Dementia (2019)