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Cdk-activating kinase complex is a component of human transcription factor TFIIH.

机译:Cdk激活激酶复合物是人类转录因子TFIIH的组成部分。

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Transcription factor IIH (TFIIH) contains a kinase capable of phosphorylating the carboxy-terminal domain (CTD) of the largest subunit of RNA polymerase II (RNAPII). Here we report the identification of the Cdk-activating kinase (Cak) complex (Cdk7 and cyclin H) as a component of TFIIH after extensive purification of TFIIH by chromatography. We find that affinity-purified antibodies directed against cyclin H inhibit TFIIH-dependent transcription and that both cyclin H and Cdk7 antibodies inhibit phosphorylation of the CTD of the largest subunit of the RNAPII in the preinitiation complex. Cak is present in at least two distinct complexes, TFIIH and a smaller complex that is unable to phosphorylate RNAPII in the preinitiation complex. Both Cak complexes, as well as recombinant Cak, phosphorylate a CTD peptide. Finally, TFIIH was shown to phosphorylate both Cdc2 and Cdk2, suggesting that there could be a link between transcription and the cell cycle machinery.
机译:转录因子IIH(TFIIH)包含一种能够磷酸化RNA聚合酶II(RNAPII)最大亚基的羧基末端结构域(CTD)的激酶。在这里,我们报告了通过色谱法对TFIIH进行广泛纯化后,鉴定出Cdk活化激酶(Cak)复合物(Cdk7和细胞周期蛋白H)作为TFIIH的组成部分。我们发现针对细胞周期蛋白H的亲和纯化抗体抑制TFIIH依赖性转录,并且细胞周期蛋白H和Cdk7抗体均抑制预初始化复合物中RNAPII最大亚基CTD的磷酸化。 Cak存在于至少两种不同的复合物中,即TFIIH和一种较小的复合物中,该复合物无法使预起始复合物中的RNAPII磷酸化。 Cak复合物和重组Cak都可磷酸化CTD肽。最后,显示TFIIH可以磷酸化Cdc2和Cdk2,表明转录和细胞周期机制之间可能存在联系。

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