Redundant roles of Srs2 helicase and replication checkpoint in survival and rDNA maintenance in Schizosaccharomyces pombe.
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 Schizosaccharomyces 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 (approximately 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.
Yasuhira
Shinji
S
Division of Radiation Life Science, Research Reactor Institute, Kyoto University, 2-1010 Asashironishi, Kumatori-Cho, Sennan-Gun, Osaka, 590-0494, Japan. shinji@rri.kyoto-u.ac.jp
D001483:Base Sequence; D002453:Cell Cycle; D004249:DNA Damage; D004265:DNA Helicases / Q000235:genetics* / Q000378:metabolism*; D004261:DNA Replication; D004271:DNA, Fungal / Q000235:genetics* / Q000378:metabolism* / Q000528:radiation effects; D004275:DNA, Ribosomal / Q000235:genetics* / Q000378:metabolism*; D005720:Gamma Rays; D018628:Gene Dosage; D005800:Genes, Fungal; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010641:Phenotype; D051135:Rad51 Recombinase / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D012568:Schizosaccharomyces / Q000166:cytology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics* / Q000378:metabolism*; D013696:Temperature
D004271:DNA, Fungal; D004275:DNA, Ribosomal; C084232:RHP51 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; D051135:Rad51 Recombinase; D004265:DNA Helicases; C468289:Srs2 protein, S pombe