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First published online 15 September 2009
doi: 10.1242/jcs.046508

Journal of Cell Science 122, 3638-3643 (2009)
Published by The Company of Biologists 2009
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Research Article

Schizosaccharomyces pombe Cds1Chk2 regulates homologous recombination at stalled replication forks through the phosphorylation of recombination protein Rad60

Izumi Miyabe1,2, Takashi Morishita1,2, Hideo Shinagawa2,3 and Antony M. Carr1,*

1 Genome Damage and Stability Centre, University of Sussex, Brighton BN1 9RQ, UK
2 Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan
3 BioAcademia, 7-7-18 Saito-Asagi, Ibaraki, Osaka 567-0085, Japan

* Author for correspondence (a.m.carr{at}susssex.ac.uk)

Accepted 28 April 2009

The Schizosaccharomyces pombe rad60 gene is essential for cell growth and is involved in repairing DNA double-strand breaks. Rad60 physically interacts with, and is functionally related to, the structural maintenance of chromosomes 5 and 6 protein complex (Smc5/6). Rad60 is phosphorylated in response to hydroxyurea (HU)-induced DNA replication arrest in a Cds1Chk2-dependent manner. Rad60 localizes in nucleus in unchallenged cells, but becomes diffused throughout the cell in response to HU. To understand the role of Rad60 phosphorylation, we mutated the putative phosphorylation target motifs of Cds1Chk2 and have identified two Cds1Chk2 target residues responsible for Rad60 dispersal in response to HU. We show that the phosphorylation-defective rad60 mutation partially suppresses HU sensitivity and the elevated recombination frequency of smc6-X. Our data suggest that Rad60 phosphorylation is required to regulate homologous recombination at stalled replication forks, probably by regulating Smc5/6.

Key words: Checkpoint, DNA repair, SMC5/6


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