Sun Z W, Hampsey M
Department of Biochemistry and Molecular Biology, Louisiana State University Medical Center, Shreveport 71130-3932, USA.
Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3127-31. doi: 10.1073/pnas.92.8.3127.
Mutations in the Saccharomyces cerevisiae SSU71 gene were isolated as suppressors of a transcription factor TFIIB defect that confers both a cold-sensitive growth defect and a downstream shift in transcription start-site selection at the cyc1 locus. The ssu71-1 suppressor not only suppresses the conditional phenotype but also restores the normal pattern of transcription initiation at cyc1. In addition, the ssu71-1 suppressor confers a heat-sensitive phenotype that is dependent upon the presence of the defective form of TFIIB. Molecular and genetic analysis of the cloned SSU71 gene demonstrated that SSU71 is a single-copy essential gene encoding a highly charged protein with a molecular mass of 82,194 daltons. Comparison of the deduced Ssu71 amino acid sequence with the protein data banks revealed significant similarity to RAP74, the larger subunit of the human general transcription factor TFIIF. Moreover, Ssu71 is identical to p105, a component of yeast TFIIF. Taken together, these data demonstrate a functional interaction between TFIIB and the large subunit of TFIIF and that this interaction can affect start-site selection in vivo.
酿酒酵母SSU71基因的突变体是作为转录因子TFIIB缺陷的抑制子分离得到的,该缺陷会导致冷敏感生长缺陷以及cyc1基因座转录起始位点选择的下游偏移。ssu71-1抑制子不仅能抑制这种条件性表型,还能恢复cyc1处转录起始的正常模式。此外,ssu71-1抑制子赋予了一种热敏感表型,该表型依赖于缺陷形式的TFIIB的存在。对克隆的SSU71基因进行分子和遗传分析表明,SSU71是一个单拷贝必需基因,编码一种分子量为82,194道尔顿的高电荷蛋白。将推导的Ssu71氨基酸序列与蛋白质数据库进行比较,发现它与人类通用转录因子TFIIF的较大亚基RAP74有显著相似性。此外,Ssu71与酵母TFIIF的一个组分p105相同。综上所述,这些数据证明了TFIIB与TFIIF的大亚基之间存在功能相互作用,并且这种相互作用会影响体内的起始位点选择。