Recruitment of RecA homologs Dmc1p and Rad51p to the double-strand break repair site initiated by meiosis-specific endonuclease VDE (PI-SceI) |
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Authors: | Tomoyuki Fukuda Yoshikazu Ohya |
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Institution: | (1) Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Bldg. FSB-101, 5-1-5 Kashiwanoha Kashiwa, 277-8562 Chiba Prefecture, Japan;(2) Present address: Genetic Dynamics Research Unit Laboratory, RIKEN, 2-1 Hirosawa Wako, 351-0198 Saitama, Japan |
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Abstract: | During meiosis, VDE (PI-SceI), a homing endonuclease in Saccharomyces cerevisiae, introduces a double-strand break (DSB) at its recognition sequence and induces homologous recombinational repair, called
homing. Meiosis-specific RecA homolog Dmc1p, as well as mitotic RecA homolog Rad51p, acts in the process of meiotic recombination,
being required for strand invasion and exchange. In this study, recruitment of Dmc1p and Rad51p to the VDE-induced DSB repair
site is investigated by chromatin immunoprecipitation assay. It is revealed that Dmc1p and Rad51p are loaded to the repair
site in an independent manner. Association of Rad51p requires other DSB repair proteins of Rad52p, Rad55p, and Rad57p, while
loading of Dmc1p is facilitated by the different protein, Sae3p. Absence of Tid1p, which can bind both RecA homologs, appears
specifically to cause an abnormal distribution of Dmc1p. Lack of Hop2, Mnd1p, and Sae1p does not impair recruitment of both
RecA homologs. These findings reveal the discrete functions of each strand invasion protein in VDE-initiated homing, confirm
the similarity between VDE-initiated homing and Spo11p-initiated meiotic recombination, and demonstrate the availability of
VDE-initiated homing for the study of meiotic recombination. |
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Keywords: | Meiotic recombination RecA homolog Rad51p Dmc1p VDE |
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