Xu Q, Singer R A, Johnston G C
Department of Microbiology, Dalhousie University, Halifax, Nova Scotia, Canada.
Mol Cell Biol. 1995 Nov;15(11):6025-35. doi: 10.1128/MCB.15.11.6025.
The Cdc68 protein is required for the transcription of a variety of genes in the yeast Saccharomyces cerevisiae. In a search for proteins involved in the activity of the Cdc68 protein, we identified four suppressor genes in which mutations reverse the temperature sensitivity caused by the cdc68-1 allele. We report here the molecular characterization of mutations in one suppressor gene, the previously identified SUG1 gene. The Sug1 protein has been implicated in both transcriptional regulation and proteolysis. sug1 suppressor alleles reversed most aspects of the cdc68-1 mutant phenotype but did not suppress the lethality of a cdc68 null allele, indicating that sug1 suppression is by restoration of Cdc68 activity. Our evidence suggests that suppression by sug1 is unlikely to be due to increased stability of mutant Cdc68 protein, despite the observation that Sug1 affected proteolysis of mutant Cdc68. We report here that attenuated Sug1 activity strengthens mutant Cdc68 activity, whereas increased Sug1 activity further inhibits enfeebled Cdc68 activity, suggesting that Sug1 antagonizes the activator function of Cdc68 for transcription. Consistent with this hypothesis, we find that Sug1 represses transcription in vivo.
Cdc68蛋白是酿酒酵母中多种基因转录所必需的。在寻找参与Cdc68蛋白活性的蛋白质的过程中,我们鉴定出了四个抑制基因,其中的突变可逆转由cdc68-1等位基因引起的温度敏感性。我们在此报告一个抑制基因(先前鉴定的SUG1基因)中突变的分子特征。Sug1蛋白与转录调控和蛋白水解均有关联。sug1抑制等位基因逆转了cdc68-1突变体表型的大多数方面,但并未抑制cdc68无效等位基因的致死性,这表明sug1抑制是通过恢复Cdc68活性实现的。我们的证据表明,尽管观察到Sug1影响突变型Cdc68蛋白的蛋白水解,但sug1抑制不太可能是由于突变型Cdc68蛋白稳定性增加所致。我们在此报告,减弱的Sug1活性会增强突变型Cdc68活性,而增加的Sug1活性会进一步抑制衰弱的Cdc68活性,这表明Sug1拮抗Cdc68的转录激活功能。与该假设一致,我们发现Sug1在体内抑制转录。