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首页> 外文期刊>Molecular and Cellular Biology >Identifying a species-specific region of yeast TF11B in vivo.
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Identifying a species-specific region of yeast TF11B in vivo.

机译：在体内鉴定酵母TF11B的物种特异性区域。

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The general transcription factor IIB (TFIIB) is required for RNA polymerase II transcription in eukaryotes. It provides a physical link between the TATA-binding protein (TBP) and the RNA polymerase and is a component previously suggested to respond to transcriptional activators in vitro. In this report, we compare the yeast (Saccharomyces cerevisiae) and human forms of the protein in yeast cells to study their functional differences. We demonstrate that human TFIIB fails to functionally replace yeast TFIIB in yeast cells. By analyzing various human-yeast hybrid TFIIB molecules, we show that a 14-amino-acid region at the amino terminus of the first repeat of yeast TFIIB plays an important role in determining species specificity in vivo. In addition, we identify four amino acids in this region that are critical for an amphipathic helix unique to yeast TFIIB. By site-directed mutagenesis analyses we demonstrate that these four amino acids are important for yeast TFIIB's activity in vivo. Finally, we show that mutations in the species-specific region of yeast TFIIB can differentially affect the expression of genes activated by different activators in vivo. These results provide strong evidence suggesting that yeast TFIIB is involved in the process of transcriptional activation in living cells.
机译：真核生物中RNA聚合酶II转录需要通用转录因子IIB（TFIIB）。它提供了TATA结合蛋白（TBP）和RNA聚合酶之间的物理联系，并且是以前建议在体外对转录激活因子起反应的组分。在本报告中，我们比较了酵母细胞中的酵母（Saccharomyces cerevisiae）和人形式的蛋白质，以研究其功能差异。我们证明了人类TFIIB未能在功能上替代酵母细胞中的酵母TFIIB。通过分析各种人与酵母杂交的TFIIB分子，我们表明酵母TFIIB的第一个重复序列的氨基末端的14个氨基酸区域在确定体内物种特异性中起重要作用。此外，我们确定了该区域中的四个氨基酸，这些氨基酸对于酵母TFIIB特有的两亲性螺旋至关重要。通过定点诱变分析，我们证明了这四个氨基酸对于酵母TFIIB的体内活性很重要。最后，我们表明酵母TFIIB的物种特异性区域中的突变可以差异地影响体内不同激活剂激活的基因的表达。这些结果提供了强有力的证据，表明酵母TFIIB参与了活细胞中转录激活的过程。

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