Abstract
Srs2 helicase is believed to function as an anti-recombinase by resolving inappropriate Rad51-DNA filament. We found synthetic lethality or poor growth of srs2 with rad3 or mrc1 in Schizossacharomyces pombe. Lethality may result from a defect in non-checkpoint function of Rad3 or Mrc1 in the absence of Srs2, because srs2∆ rad9∆, srs2∆ chk1∆ cds1∆ or srs2∆ mrc1-14A (non-phosphorylatable mrc1 allele) did not show significant growth impairment. Notably, the inactivation of rhp51/RAD51 or rad22/RAD52 failed to rescue the growth, suggesting that events that impose lethality are independent of homologous recombination. Incubation of the conditional srs2∆ rad3 ts cells at restrictive temperature led not only to a viability decrease but also to a remarkable shortening of rDNA clusters (~100 copies). As opposed to the growth defect, shortening of rDNA clusters was also observed in srs2∆ rad9∆, srs2∆ chk1∆ cds1∆ or srs2∆ mrc1-14A, indicating that proper replication checkpoint signaling is critical for rDNA maintenance. Activation of Chk1 in the unchallenged mrc1-14A srs2∆ cells implies a certain level of spontaneous fork damage that might be the cause for rDNA instability. The data suggest that redundant functions of Srs2 and checkpoint proteins are essential for two independent aspects of genome maintenance.
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Acknowledgments
We thank A. M. Carr, T. Kelly, A. Matsuura, N. Rhind, P. Russell, K. Tanaka, and M. Yanagida for providing strains and plasmids, and H. Iwasaki for assistance in sequencing. This research was partly supported by the Ministry of Education, Science, Sports and Culture, Grant-in-Aid for Encouragement of Young Scientists, No. 12780510, 2000.
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Communicated by A. Aguilera.
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Yasuhira, S. Redundant roles of Srs2 helicase and replication checkpoint in survival and rDNA maintenance in Schizosaccharomyces pombe . Mol Genet Genomics 281, 497–509 (2009). https://doi.org/10.1007/s00438-009-0426-x
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DOI: https://doi.org/10.1007/s00438-009-0426-x