Abstract
DNA double-strand breaks (DSBs) initiate meiotic recombination in Schizosaccharomyces pombe and in other organisms. The Rec12 protein catalyzes the formation of these DSBs in concert with a multitude of accessory proteins the role of which in this process remains to be discovered. In an all-to-all yeast two-hybrid matrix analysis, we discovered new interactions among putative members of the meiotic recombination initiation complex. We found that Rec7, an axial-element associated protein with homologies to Saccharomyces cerevisiae Rec114, is interacting with Rec24. Rec7 and Rec24 also co-immunoprecipitate in S. pombe during meiosis. An amino acid change in a conserved, C-terminal phenylalanine in Rec7, F325A interrupts the interaction with Rec24. Moreover, rec7F325A shows a recombination deficiency comparable to rec7Δ. Another interaction was detected between Rec12 and Rec14, the orthologs of which in S. cerevisiae Spo11 and Ski8 interact accordingly. Amino acid changes Rec12Q308A and Rec12R309A disrupt the interaction with Rec14, like the according amino acid changes Spo11Q376A and Spo11RE377AA loose the interaction with Ski8. Both amino acid changes in Rec12 reveal a recombination deficient rec12 − phenotype. We propose that both Rec7–Rec24 and Rec12–Rec14 form subcomplexes of the meiotic recombination initiation complex.
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Acknowledgments
We thank Marc-David Ruepp and Kathleen Gould for plasmids, Valerie Borde, Cristina Martin-Castellanos, Jurai Gregan, and Franz Klein for helpful discussions. Our acknowledgements for S. cerevisiae strains and Y2H plasmids go to T. N. Davis, grant P41 RR11823 from the National Center for Research Resources at the National Institutes of Health. The study was supported by the Swiss National Science Foundation to JK and KL. KL was supported by the UniBern Forschungstiftung and SS by a short-term fellowship of the Boehringer Ingelheim Fonds.
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Communicated by M. Yamamoto.
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Steiner, S., Kohli, J. & Ludin, K. Functional interactions among members of the meiotic initiation complex in fission yeast. Curr Genet 56, 237–249 (2010). https://doi.org/10.1007/s00294-010-0296-0
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DOI: https://doi.org/10.1007/s00294-010-0296-0