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Molecular and Cellular Biology, June 2005, p. 4377-4387, Vol. 25, No. 11
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.11.4377-4387.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Fission Yeast Rad51 and Dmc1, Two Efficient DNA Recombinases Forming Helical Nucleoprotein Filaments

Synthia Sauvageau,1 Alicja Z. Stasiak,2 Isabelle Banville,1 Mickaël Ploquin,1 Andrzej Stasiak,2 and Jean-Yves Masson1*

Genome Stability Laboratory, Laval University Cancer Research Center, Hôtel-Dieu de Québec, 9 McMahon, Québec City, Quebec G1R 2J6, Canada,1 Laboratoire d'Analyse Structurale, Université de Lausanne, 1015 Lausanne, Switzerland2

Received 28 January 2005/ Returned for modification 2 March 2005/ Accepted 8 March 2005

Homologous recombination is important for the repair of double-strand breaks during meiosis. Eukaryotic cells require two homologs of Escherichia coli RecA protein, Rad51 and Dmc1, for meiotic recombination. To date, it is not clear, at the biochemical level, why two homologs of RecA are necessary during meiosis. To gain insight into this, we purified Schizosaccharomyces pombe Rad51 and Dmc1 to homogeneity. Purified Rad51 and Dmc1 form homo-oligomers, bind single-stranded DNA preferentially, and exhibit DNA-stimulated ATPase activity. Both Rad51 and Dmc1 promote the renaturation of complementary single-stranded DNA. Importantly, Rad51 and Dmc1 proteins catalyze ATP-dependent strand exchange reactions with homologous duplex DNA. Electron microscopy reveals that both S. pombe Rad51 and Dmc1 form nucleoprotein filaments. Rad51 formed helical nucleoprotein filaments on single-stranded DNA, whereas Dmc1 was found in two forms, as helical filaments and also as stacked rings. These results demonstrate that Rad51 and Dmc1 are both efficient recombinases in lower eukaryotes and reveal closer functional and structural similarities between the meiotic recombinase Dmc1 and Rad51. The DNA strand exchange activity of both Rad51 and Dmc1 is most likely critical for proper meiotic DNA double-strand break repair in lower eukaryotes.

* Corresponding author. Mailing address: Genome Stability Laboratory, Laval University Cancer Research Center, Hôtel-Dieu de Québec, 9 McMahon, Quebec City, Quebec G1R 2J6, Canada. Phone: 418-525-4444, ext. 15154. Fax: 418-691-5439. E-mail: Jean-Yves.Masson{at}crhdq.ulaval.ca.

Molecular and Cellular Biology, June 2005, p. 4377-4387, Vol. 25, No. 11
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.11.4377-4387.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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