Cho Nam-Chul, Kang Hyun-Jung, Lim Hye-Won, Kim Byung-Chul, Park Eun-Hee, Lim Chang-Jin
Division of Life Sciences, Kangwon National University, College of Natural Sciences, Chuncheon, Korea.
Can J Microbiol. 2006 Dec;52(12):1261-5. doi: 10.1139/w06-081.
To elicit the physiological roles of Pbh1, a baculoviral IAP repeat (BIR) domain-containing protein, in Schizosaccharomyces pombe, we investigated if Pbh1 expression is regulated by stress. The upstream region (1221 bp) of the pbh1 gene was fused into the promoterless beta-galactosidase gene of the shuttle vector YEp367R, and the resultant fusion plasmid was named pPbh04. The synthesis of beta-galactosidase from the pbh1-lacZ fusion gene was markedly enhanced by sodium nitroprusside (SNP) generating nitric oxide. The basal expression of the pbh1 gene required the presence of Pap1. Pap1 also mediated the induction of the pbh1 gene by SNP and nitrogen starvation. Pap1-dependent induction of the pbh1 gene by SNP was confirmed by the enhanced level of the pbh1 mRNA in Pap1-positive cells but not in Pap1-negative cells. Taken together, it was demonstrated that the pbh1 genes are positively regulated by nitrosative and nitrogen starvation stresses in Pap1-dependent manner.
为了探究含有杆状病毒IAP重复序列(BIR)结构域的蛋白Pbh1在粟酒裂殖酵母中的生理作用,我们研究了Pbh1的表达是否受应激调控。将pbh1基因的上游区域(1221 bp)与穿梭载体YEp367R的无启动子β-半乳糖苷酶基因融合,所得融合质粒命名为pPbh04。产生一氧化氮的硝普钠(SNP)显著增强了pbh1-lacZ融合基因的β-半乳糖苷酶合成。pbh1基因的基础表达需要Pap1的存在。Pap1还介导了SNP和氮饥饿对pbh1基因的诱导。通过Pap1阳性细胞中pbh1 mRNA水平升高而Pap1阴性细胞中未升高,证实了SNP对pbh1基因的Pap1依赖性诱导。综上所述,证明了pbh1基因在Pap1依赖性方式下受到亚硝化和氮饥饿应激的正调控。