Protective role and regulation of Rad9 from the fission yeast Schizosaccharomyces pombe.
To assess novel cellular roles and regulation of Rad9 in the fission yeast Schizosaccharomyces pombe, the full-length rad9 gene was cloned into the shuttle vector pRS316, generating pYFRad9. The rad9 mRNA level was significantly increased in the S. pombe cells harboring the plasmid pYFRad9, suggesting that the cloned rad9 gene is functioning. The S. pombe cells harboring pYFRad9 showed higher survival in the minimal media containing nitric oxide (NO)-generating sodium nitroprusside (SNP, 20 muM) and no nitrogen than the vector control cells. SNP and nitrogen starvation notably enhanced the synthesis of beta-galactosidase from the rad9-lacZ fusion gene in the Pap1-positive cells but not in the Pap1-negative cells. The rad9 mRNA level, detected by semi-quantitative reverse transcriptase (RT)-PCR, was elevated in the Pap1-positive cells but not in the Pap1-negative cells by SNP and nitrogen starvation. It was also increased only in the Pap1-positive cells by diethylmaleate, which activates Pap1. Collectively, the results imply that Rad9 plays a protective role against nitrosative and nutritional stress and is positively regulated by NO and nitrogen starvation in a Pap1-dependent manner.
Kang
Min-Hee
MH
Division of Life Sciences and Research Institute of Life Sciences, College of Natural Sciences, Kangwon National University, Chuncheon, Korea.
Park
Eun-Hee
EH
Lim
Chang-Jin
CJ
D050976:Basic-Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D004271:DNA, Fungal / Q000032:analysis; D015966:Gene Expression Regulation, Fungal; D018869:Heat-Shock Response; D016350:Leucine Zippers; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000378:metabolism / Q000494:pharmacology*; D009584:Nitrogen / Q000378:metabolism / Q000494:pharmacology*; D000073718:Pancreatitis-Associated Proteins; D012568:Schizosaccharomyces / Q000235:genetics / Q000254:growth & development / Q000378:metabolism* / Q000502:physiology; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA
D050976:Basic-Leucine Zipper Transcription Factors; D018797:Cell Cycle Proteins; D004271:DNA, Fungal; D000073718:Pancreatitis-Associated Proteins; C105096:Pap1 protein, S pombe; C000612141:REG3A protein, human; D029702:Schizosaccharomyces pombe Proteins; C069930:rad9 protein; D009569:Nitric Oxide; D009584:Nitrogen