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Molecular and Cellular Biology, May 2002, p. 3537-3548, Vol. 22, No. 10
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.10.3537-3548.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

The Schizosaccharomyces pombe rad60 Gene Is Essential for Repairing Double-Strand DNA Breaks Spontaneously Occurring during Replication and Induced by DNA-Damaging Agents

Takashi Morishita,1 Yasuhiro Tsutsui,1 Hiroshi Iwasaki,1,2,{dagger} and Hideo Shinagawa1*

Department of Molecular Microbiology, Research Institute for Microbial Diseases, Osaka University,1 PRESTO JST, Suita, Osaka 565-0871, Japan2

Received 13 December 2001/ Returned for modification 23 January 2002/ Accepted 22 February 2002

To identify novel genes involved in DNA double-strand break (DSB) repair, we previously isolated Schizosaccharomyces pombe mutants which are hypersensitive to methyl methanesulfonate (MMS) and synthetic lethals with rad2. This study characterizes one of these mutants, rad60-1. The gene that complements the MMS sensitivity of this mutant was cloned and designated rad60. rad60 encodes a protein with 406 amino acids which has the conserved ubiquitin-2 motif found in ubiquitin family proteins. rad60-1 is hypersensitive to UV and {gamma} rays, epistatic to rhp51, and defective in the repair of DSBs caused by {gamma}-irradiation. The rad60-1 mutant is also temperature sensitive for growth. At the restrictive temperature (37°C), rad60-1 cells grow for several divisions and then arrest with 2C DNA content; the arrested cells accumulate DSBs and have a diffuse and often aberrantly shaped nuclear chromosomal domain. The rad60-1 mutant is a synthetic lethal with rad18-X, and expression of wild-type rad60 from a multicopy plasmid partially suppresses the MMS sensitivity of rad18-X cells. rad18 encodes a conserved protein of the structural maintenance of chromosomes (SMC) family (A. R. Lehmann, M. Walicka, D. J. Griffiths, J. M. Murray, F. Z. Watts, S. McCready, and A. M. Carr, Mol. Cell. Biol. 15:7067-7080, 1995). These results suggest that S. pombe Rad60 is required to repair DSBs, which accumulate during replication, by recombination between sister chromatids. Rad60 may perform this function in concert with the SMC protein Rad18.

* Corresponding author. Mailing address: Department of Molecular Microbiology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan. Phone: 81-6-6879-8317. Fax: 81-6-6879-8320. E-mail: shinagaw{at}biken.osaka-u.ac.jp.

{dagger} Present address: Science of Biological Supermolecular Systems, Graduate School of Integrated Science, Yokohama City University, Tsurumi-ku, Yokohama, Kanagawa, Japan.

Molecular and Cellular Biology, May 2002, p. 3537-3548, Vol. 22, No. 10
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.10.3537-3548.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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