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首页> 外文期刊>Molecular and Cellular Biology >Identification of RTF1, a novel gene important for TATA site selection by TATA box-binding protein in Saccharomyces cerevisiae.
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Identification of RTF1, a novel gene important for TATA site selection by TATA box-binding protein in Saccharomyces cerevisiae.

机译：RTF1的鉴定，这对酿酒酵母中TATA盒结合蛋白对TATA位点选择至关重要的新基因。

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Interaction of the TATA box-binding protein (TBP) with promoters of RNA polymerase II-transcribed genes is an early and essential step in mRNA synthesis. Previous studies have demonstrated that the rate-limiting binding of TBP to a TATA element can be influenced by transcriptional regulatory proteins. To identify additional factors that may regulate DNA binding by TBP in vivo, we performed a genetic selection for extragenic suppressors of a yeast TBP mutant that exhibits altered and relaxed DNA binding specificity. This analysis has led to the discovery of a previously unidentified gene, RTF1. The original rtf1 suppressor mutation, which encodes a single amino acid change in Rtf1, and an rtf1 null allele suppress the effects of the TBP specificity mutant by altering transcription initiation. Differences in the patterns of transcription initiation in these strains strongly suggest that the rtf1 missense mutation is distinct from a simple loss-of-function allele. The results of genetic crosses indicate that suppression of TBP mutants by mutations in RTF1 occurs in an allele-specific fashion. In a strain containing wild-type TBP, the rtf1 null mutation suppresses the transcriptional effects of a Ty delta insertion mutation in the promoter of the HIS4 gene, a phenotype also conferred by the TBP altered-specificity mutant. Finally, as shown by indirect immunofluorescence experiments, Rtf1 is a nuclear protein. Taken together, our findings suggest that Rtf1 either directly or indirectly regulates the DNA binding properties of TBP and, consequently, the relative activities of different TATA elements in vivo.
机译：TATA盒结合蛋白（TBP）与RNA聚合酶II转录基因的启动子的相互作用是mRNA合成的早期且必不可少的步骤。以前的研究表明，转录调节蛋白会影响TBP与TATA元件的限速结合。为了确定其他可能在体内调节TBP与DNA结合的因素，我们对酵母TBP突变体的外源抑制子进行了遗传选择，该突变体表现出改变的DNA结合特异性和松弛的DNA结合特异性。该分析导致发现了一个以前未鉴定的基因RTF1。原始的rtf1抑制子突变编码Rtf1中的单个氨基酸变化，而rtf1无效等位基因通过改变转录起始来抑制TBP特异性突变体的作用。这些菌株中转录起始模式的差异强烈表明，rtf1错义突变不同于简单的功能丧失等位基因。遗传杂交的结果表明，RTF1突变对TBP突变的抑制以等位基因特异性的方式发生。在含有野生型TBP的菌株中，rtf1无效突变抑制了HIS4基因启动子中Ty delta插入突变的转录作用，该表型也由TBP改变特异性突变体赋予。最后，如间接免疫荧光实验所示，Rtf1是核蛋白。两者合计，我们的发现表明Rtf1直接或间接调节TBP的DNA结合特性，并因此调节体内不同TATA元件的相对活性。

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