Srs2 helicase prevents the formation of toxic DNA damage during late prophase I of yeast meiosis
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Proper repair of double-strand breaks (DSBs) is key to ensure proper chromosome segregation. In this study, we found that the deletion of the SRS2 gene, which encodes a DNA helicase necessary for the control of homologous recombination, induces aberrant chromosome segregation during budding yeast meiosis. This abnormal chromosome segregation in srs2 cells accompanies the formation of a novel DNA damage induced during late meiotic prophase I. The damage may contain long stretches of single-stranded DNAs (ssDNAs), which lead to aggregate formation of a ssDNA binding protein, RPA, and a RecA homolog, Rad51, as well as other recombination proteins inside of the nuclei, but not that of a meiosis-specific Dmc1. The Rad51 aggregate formation in the srs2 mutant depends on the initiation of meiotic recombination and occurs in the absence of chromosome segregation. Importantly, as an early recombination intermediate, we detected a thin bridge of Rad51 between two Rad51 foci in the srs2 mutant, which is rarely seen in wild type. These might be cytological manifestation of the connection of two DSB ends and/or multi-invasion. The DNA damage with Rad51 aggregates in the srs2 mutant is passed through anaphases I and II, suggesting the absence of DNA damage-induced cell cycle arrest after the pachytene stage. We propose that Srs2 helicase resolves early protein-DNA recombination intermediates to suppress the formation of aberrant lethal DNA damage during late prophase I.
KeywordsSrs2 Rad51 Dmc1 Meiotic recombination
We are grateful for Drs. Alastair Goldman and Michael Lichten for sharing unpublished results prior to publication. We thank Dr. Neil Hunter (UC, Davis) for pCLB2-SGS1 yeast and Dr. Andreas Hochwagen (New York University) for the anchor-away yeast strains. We thank Ms. H. Matsumoto, S. Hashimoto, C. Watanabe, and H. Wakabayashi for excellent technical assistance.
H.S., M.S., and A.S. designed the experiments. H.S., H.S.M.S., Y.F., K.C., L.P., and A.S. performed experiments. M.S. provided reagents. H.S., H.S.M.S., and A.S. analyzed the data. A.S. prepared manuscripts with help by H.S., H.S.M.S., S.G., and M.S.
This work was supported by the Japanese Society for Promotion of Science (JSPS) KAKENHI Grant Numbers 22125001, 22125002, 15H05973, and 16H04742 to A.S.; 21770005 to H.S.; 15H05973, M.S. H.S.M.S. was supported by Institute for Protein Research.
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