Rowley A, Singer R A, Johnston G C
Department of Microbiology, Dalhousie University, Halifax, Nova Scotia, Canada.
Mol Cell Biol. 1991 Nov;11(11):5718-26. doi: 10.1128/mcb.11.11.5718-5726.1991.
The cell cycle of the budding yeast Saccharomyces cerevisiae has been investigated through the study of conditional cdc mutations that specifically affect cell cycle performance. Cells bearing the cdc68-1 mutation (J. A. Prendergast, L. E. Murray, A. Rowley, D. R. Carruthers, R. A. Singer, and G. C. Johnston, Genetics 124:81-90, 1990) are temperature sensitive for the performance of the G1 regulatory event, START. Here we describe the CDC68 gene and present evidence that the CDC68 gene product functions in transcription. CDC68 encodes a 1,035-amino-acid protein with a highly acidic and serine-rich carboxyl terminus. The abundance of transcripts from several unrelated genes is decreased in cdc68-1 mutant cells after transfer to the restrictive temperature, while at least one transcript, from the HSP82 gene, persists in an aberrant fashion. Thus, the cdc68-1 mutation has both positive and negative effects on gene expression. Our findings complement those of Malone et al. (E. A. Malone, C. D. Clark, A. Chiang, and F. Winston, Mol. Cell. Biol. 11:5710-5717, 1991), who have independently identified the CDC68 gene (as SPT16) as a transcriptional suppressor of delta-insertion mutations. Among transcripts that rapidly become depleted in cdc68-1 mutant cells are those of the G1 cyclin genes CLN1, CLN2, and CLN3/WHI1/DAF1, whose activity has been previously shown to be required for the performance of START. The decreased abundance of cyclin transcripts in cdc68-1 mutant cells, coupled with the suppression of cdc68-1-mediated START arrest by the CLN2-1 hyperactive allele of CLN2, shows that the CDC68 gene affects START through cyclin gene expression.
通过对特异性影响细胞周期进程的条件性cdc突变的研究,对出芽酵母酿酒酵母的细胞周期进行了调查。携带cdc68-1突变的细胞(J. A. 普伦德加斯特、L. E. 默里、A. 罗利、D. R. 卡拉瑟斯、R. A. 辛格和G. C. 约翰斯顿,《遗传学》124:81-90,1990)对G1调节事件START的进程具有温度敏感性。在此,我们描述了CDC68基因,并提供证据表明CDC68基因产物在转录中发挥作用。CDC68编码一种含有1035个氨基酸的蛋白质,其羧基末端具有高度酸性且富含丝氨酸。在转移到限制温度后,cdc68-1突变细胞中几个不相关基因的转录本丰度降低,而来自HSP82基因的至少一种转录本以异常方式持续存在。因此,cdc68-1突变对基因表达既有正向影响也有负向影响。我们的发现补充了马龙等人(E. A. 马龙、C. D. 克拉克、A. 蒋和F. 温斯顿,《分子细胞生物学》11:5710-5717,1991)的研究结果,他们独立地将CDC68基因(作为SPT16)鉴定为δ插入突变的转录抑制因子。在cdc68-1突变细胞中迅速减少的转录本包括G1周期蛋白基因CLN1、CLN2和CLN3/WHI1/DAF1的转录本,其活性先前已被证明是START进程所必需的。cdc68-1突变细胞中周期蛋白转录本丰度的降低,加上CLN2的CLN2-1高活性等位基因对cdc68-1介导的START停滞的抑制作用,表明CDC68基因通过周期蛋白基因表达影响START。