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首页> 外文期刊>Molecular and Cellular Biology >Association of Transcription Factor IIA with TATA Binding Protein Is Required for Transcriptional Activation of a Subset of Promoters and Cell Cycle Progression inSaccharomyces cerevisiae
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Association of Transcription Factor IIA with TATA Binding Protein Is Required for Transcriptional Activation of a Subset of Promoters and Cell Cycle Progression inSaccharomyces cerevisiae

机译：酿酒酵母子集的转录激活和细胞周期进程需要转录因子IIA与TATA结合蛋白的关联。

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The general transcription factor IIA (TFIIA) interacts with the TATA binding protein (TBP) and promoter DNA to mediate transcription activation in vitro. To determine if this interaction is generally required for activation of all class II genes in vivo, we have constructed substitution mutations in yeast TFIIA which compromise its ability to bind TBP. Substitution mutations in the small subunit of TFIIA (Toa2) at residue Y69 or W76 significantly impaired the ability of TFIIA to stimulate TBP-promoter binding in vitro. Gene replacement of wild-type TOA2 with a W76E orY69A/W76A mutant was lethal in Saccharomyces cerevisiae, while the Y69F/W76F mutant exhibited extremely slow growth at 30°C. Both the Y69A andW76A mutants were conditionally lethal at higher temperatures. Light microscopy indicated that viable toa2mutant strains accumulate as equal-size dumbbells and multibudded clumps. Transcription of the cell cycle-regulatory genesCLB1, CLB2, CLN1, andCTS1 was significantly reduced in the toa2mutant strains, while the noncycling genes PMA1 andENO2 were only modestly affected, suggesting that thesetoa2 mutant alleles disrupt cell cycle progression. The differential effect of these toa2 mutants on gene transcription was examined for a number of other genes.toa2 mutant strains supported high levels ofCUP1, PHO5, TRP3, andGAL1 gene activation, but the constitutive expression ofDED1 was significantly reduced. Activator-induced start site expression for HIS3, GAL80,URA1, and URA3 promoters was defective intoa2 mutant strains, suggesting that the TFIIA-TBP complex is important for promoters which require an activator-dependent start site selection from constitutive to regulated expression. We present evidence to indicate that transcription defects in toa2mutants can be both activator and promoter dependent. These results suggest that the association of TFIIA with TBP regulates activator-induced start site selection and cell cycle progression inS. cerevisiae.
机译：通用转录因子IIA（TFIIA）与TATA结合蛋白（TBP）和启动子DNA相互作用，以介导体外转录激活。为了确定在体内激活所有II类基因通常是否需要这种相互作用，我们在酵母TFIIA中构建了替代突变，其破坏了其结合TBP的能力。 TFIIA小亚基（Toa2）的Y69或W76残基处的取代突变显着削弱了TFIIA体外刺激TBP-启动子结合的能力。用 W76E 或 Y69A / W76A 突变体对野生型 TOA2 的基因替换对啤酒酵母（Saccharomyces cerevisiae）具有致命性，而 Y69F / W76F 突变体在30°C时生长极为缓慢。 Y69A 和 W76A 突变体均在较高温度下有条件地致死。光学显微镜表明，活的 toa2 突变株以等大小的哑铃和多预算的团块形式积累。 CLB1 ， CLB2 ， CLN1 和 CTS1 的细胞周期调控基因的转录在em> toa2 突变株，而非循环基因 PMA1 和 ENO2 仅受到中等程度的影响，表明这些 toa2 突变等位基因破坏了细胞周期进程。还检查了这些 toa2 突变体对许多其他基因的基因转录的差异作用。 toa2 突变体菌株支持高水平的 CUP1 ，< em> PHO5 ， TRP3 和 GAL1 基因激活，但 DED1 的组成型表达明显降低。 HIS3 ， GAL80 ， URA1 和 URA3 启动子的激活剂诱导的起始位点表达在中有缺陷toa2 突变菌株，表明TFIIA-TBP复合物对于需要从组成型表达到调控表达依赖激活子的起始位点选择的启动子很重要。我们提供的证据表明 toa2 突变体中的转录缺陷可能既依赖激活剂又依赖启动子。这些结果表明，TFIAA与TBP的关联调节了 S中激活剂诱导的起始位点选择和细胞周期进程。酿酒厂。

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