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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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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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Acknowledgement

This work was supported in part by a grant-in-aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan, and also assisted by Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists to T.F. We thank Satoru Nogami for critical reading of the manuscript.

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  1. Tomoyuki Fukuda

    Present address: Genetic Dynamics Research Unit Laboratory, RIKEN, 2-1 Hirosawa Wako, 351-0198, Saitama, Japan

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  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

    Tomoyuki Fukuda & Yoshikazu Ohya

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  1. Tomoyuki Fukuda
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  2. Yoshikazu Ohya
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Correspondence to Yoshikazu Ohya.

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Communicated by A. Aguilera

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Fukuda, T., Ohya, Y. Recruitment of RecA homologs Dmc1p and Rad51p to the double-strand break repair site initiated by meiosis-specific endonuclease VDE (PI-SceI). Mol Genet Genomics 275, 204–214 (2006). https://doi.org/10.1007/s00438-005-0078-4

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