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Transcriptional activation independent of TFIIH kinase and the RNA polymerase II mediator in vivo.

机译:在体内转录激活独立于TFIIH激酶和RNA聚合酶II介体。

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The carboxy-terminal domain (CTD) of the largest subunit of RNA polymerase II becomes multiply phosphorylated by protein kinases during early steps in the gene transcription cycle both in vivo and in vitro. In yeast, the major CTD kinase is a subunit of the general transcription factor TFIIH, and is encoded by an essential gene, KIN28. Although the CTD and its phosphorylation are important for transcription, in vitro studies have challenged whether CTD phosphorylation is an absolutely required step. The general importance of CTD phosphorylation by Kin28 for transcription in yeast has been suggested because, for all genes tested, transcription is inhibited at the non-permissive temperature in temperature-sensitive kin28 mutants. However, using such a mutant and a copper-inducible targeted destruction method, we show here that transcription of certain genes can be highly induced even when cells lack Kin28. We also show that transcription of these Kin28-independent genes is independent of Srb4 and Srb6, critical components of the CTD-associated transcriptional mediator complex. These results indicate that there are at least two distinct pathways for transcriptional activation: one is dependent on Kin28 and the mediator complex, and the other is not.
机译:RNA聚合酶II的最大亚基的羧基末端结构域(CTD)在体内和体外的基因转录周期的早期阶段,被蛋白激酶多重磷酸化。在酵母中,主要的CTD激酶是一般转录因子TFIIH的一个亚基,由必需基因KIN28编码。尽管CTD及其磷酸化对于转录很重要,但体外研究对CTD磷酸化是否是绝对必要的步骤提出了挑战。有人提出了Kin28的CTD磷酸化对于酵母中转录的一般重要性,因为对于所有测试的基因,在温度敏感的kin28突变体的非容许温度下,转录都受到抑制。但是,使用这种突变体和铜诱导的靶向破坏方法,我们在这里表明即使细胞缺乏Kin28,也可以高度诱导某些基因的转录。我们还显示,这些Kin28独立基因的转录独立于Srb4和Srb6,这是CTD相关转录介体复合体的关键组成部分。这些结果表明至少有两种不同的转录激活途径:一种依赖于Kin28和介体复合物,而另一种则不。

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