Stimulation of fission yeast and mouse Hop2-Mnd1 of the Dmc1 and Rad51 recombinases |
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Authors: | Ploquin Mickaël Petukhova Galina V Morneau Dany Déry Ugo Bransi Ali Stasiak Andrzej Camerini-Otero R Daniel Masson Jean-Yves |
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Affiliation: | Mickaël Ploquin, Galina V. Petukhova, Dany Morneau, Ugo Déry, Ali Bransi, Andrzej Stasiak, R. Daniel Camerini-Otero, and Jean-Yves Masson |
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Abstract: | Genetic analysis of fission yeast suggests a role for the spHop2–Mnd1 proteins in the Rad51 and Dmc1-dependent meiotic recombination pathways. In order to gain biochemical insights into this process, we purified Schizosaccharomyces pombe Hop2-Mnd1 to homogeneity. spHop2 and spMnd1 interact by co-immunoprecipitation and two-hybrid analysis. Electron microscopy reveals that S. pombe Hop2–Mnd1 binds single-strand DNA ends of 3′-tailed DNA. Interestingly, spHop2-Mnd1 promotes the renaturation of complementary single-strand DNA and catalyses strand exchange reactions with short oligonucleotides. Importantly, we show that spHop2-Mnd1 stimulates spDmc1-dependent strand exchange and strand invasion. Ca2+ alleviate the requirement for the order of addition of the proteins on DNA. We also demonstrate that while spHop2-Mnd1 affects spDmc1 specifically, mHop2 or mHop2-Mnd1 stimulates both the hRad51 and hDmc1 recombinases in strand exchange assays. Thus, our results suggest a crucial role for S. pombe and mouse Hop2-Mnd1 in homologous pairing and strand exchange and reveal evolutionary divergence in their specificity for the Dmc1 and Rad51 recombinases. |
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