Macpherson N, Measday V, Moore L, Andrews B
Department of Molecular and Medical Genetics, University of Toronto, Toronto, Ontario M55 1A8, Canada.
Genetics. 2000 Apr;154(4):1561-76. doi: 10.1093/genetics/154.4.1561.
In Saccharomyces cerevisiae, the Swi6 protein is a component of two transcription factors, SBF and MBF, that promote expression of a large group of genes in the late G1 phase of the cell cycle. Although SBF is required for cell viability, SWI6 is not an essential gene. We performed a synthetic lethal screen to identify genes required for viability in the absence of SWI6 and identified 10 complementation groups of swi6-dependent lethal mutants, designated SLM1 through SLM10. We were most interested in mutants showing a cell cycle arrest phenotype; both slm7-1 swi6Delta and slm8-1 swi6Delta double mutants accumulated as large, unbudded cells with increased 1N DNA content and showed a temperature-sensitive growth arrest in the presence of Swi6. Analysis of the transcript levels of cell cycle-regulated genes in slm7-1 SWI6 mutant strains at the permissive temperature revealed defects in regulation of a subset of cyclin-encoding genes. Complementation and allelism tests showed that SLM7 is allelic with the TAF17 gene, which encodes a histone-like component of the general transcription factor TFIID and the SAGA histone acetyltransferase complex. Sequencing showed that the slm7-1 allele of TAF17 is predicted to encode a version of Taf17 that is truncated within a highly conserved region. The cell cycle and transcriptional defects caused by taf17(slm7-1) are consistent with the role of TAF(II)s as modulators of transcriptional activation and may reflect a role for TAF17 in regulating activation by SBF and MBF.
在酿酒酵母中,Swi6蛋白是两种转录因子SBF和MBF的组成成分,这两种转录因子可促进细胞周期G1期晚期一大组基因的表达。虽然SBF是细胞存活所必需的,但SWI6不是必需基因。我们进行了一项合成致死筛选,以鉴定在缺乏SWI6时细胞存活所需的基因,并鉴定出10个swi6依赖性致死突变体的互补组,命名为SLM1至SLM10。我们对表现出细胞周期停滞表型的突变体最感兴趣;slm7-1 swi6Δ和slm8-1 swi6Δ双突变体均积累为大的、未出芽的细胞,1N DNA含量增加,并且在有Swi6存在的情况下表现出温度敏感型生长停滞。在允许温度下对slm7-1 SWI6突变体菌株中细胞周期调控基因的转录水平进行分析,发现细胞周期蛋白编码基因亚群的调控存在缺陷。互补和等位性测试表明,SLM7与TAF17基因等位,TAF17基因编码通用转录因子TFIID和SAGA组蛋白乙酰转移酶复合物的组蛋白样成分。测序表明,TAF17的slm7-1等位基因预计编码一种在高度保守区域内被截断的Taf17版本。taf17(slm7-1)引起的细胞周期和转录缺陷与TAF(II)s作为转录激活调节剂的作用一致,可能反映了TAF17在调节SBF和MBF激活方面的作用。