Yhh1p/Cft1p directly links poly(A) site recognition and RNA polymerase II transcription termination
Yhh1p/Cft1p directly links poly(A) site recognition and RNA polymerase II transcription termination
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DOI：
10.1093/emboj/cdf390
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发表时间：
2002-08-01
期刊：
影响因子：
11.4
通讯作者：
Keller, W
中科院分区：
文献类型：
--
作者：
Dichtl, B;Blank, D;Keller, W
RNA聚合酶II（pol II）的转录终止需要一个功能性的多聚腺苷酸化信号的共转录识别，但这个信号的pol II的分子机制仍然不清楚。我们表明，Yhh 1 p/Cft 1 p，酵母同源的哺乳动物AAUAAA相互作用蛋白CPSF 160，是一种RNA结合蛋白，并提供证据表明，它参与聚（A）位点识别。有趣的是，RNA结合是由预测的β螺旋桨形成重复序列组成的中心结构域介导的，这发生在具有不同细胞功能的蛋白质中。我们还发现，Yhh 1 p/Cft 1 p结合特异性磷酸化的C-末端结构域（CTD）的pol II在体外和在体内的双杂交试验。此外，转录运行分析表明，yhh 1突变体在转录终止中有缺陷，表明Yhh 1 p/Cft 1 p在转录和3 '端形成的耦合中起作用。我们认为，Yhh 1 p/Cft 1 p与RNA转录物和CTD的直接相互作用是必需的，以将poly（A）位点识别传递到延长pol II以启动转录终止。
RNA polymerase II (pol II) transcription termination requires co-transcriptional recognition of a functional polyadenylation signal, but the molecular mechanisms that transduce this signal to pol II remain unclear. We show that Yhh1p/Cft1p, the yeast homologue of the mammalian AAUAAA interacting protein CPSF 160, is an RNA-binding protein and provide evidence that it participates in poly(A) site recognition. Interestingly, RNA binding is mediated by a central domain composed of predicted beta-propeller-forming repeats, which occurs in proteins of diverse cellular functions. We also found that Yhh1p/Cft1p bound specifically to the phosphorylated C-terminal domain (CTD) of pol II in vitro and in a two-hybrid test in vivo. Furthermore, transcriptional run-on analysis demonstrated that yhh1 mutants were defective in transcription termination, suggesting that Yhh1p/Cft1p functions in the coupling of transcription and 3'-end formation. We propose that direct interactions of Yhh1p/Cft1p with both the RNA transcript and the CTD are required to communicate poly(A) site recognition to elongating pol II to initiate transcription termination.