RNA polymerase IICTD phosphopeptides compete with RNA for the interaction with Pcf11
RNA polymerase IICTD phosphopeptides compete with RNA for the interaction with Pcf11
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作者：
Hollingworth, D;Noble, CG;Ramos, A
在酿酒酵母中，裂解/多聚腺苷酸化因子Pcf 11是一个重要的调节因子，需要招募多聚腺苷酸化机制的延伸RNA聚合酶II（RNAPII），是正确的转录终止所必需的。与RNAPII的相互作用由位于Pcf 11 N-末端区域的CTD相互作用结构域（CID）介导，其以磷酸依赖性方式结合RNAPII CTD中的七肽重复序列。我们以前研究过这种蛋白质-蛋白质相互作用。我们在这里检查的CID与不同的RNA序列的相互作用，并期待来自CTD七肽重复的RNA-蛋白质相互作用的磷酸肽的效果。我们的研究结果表明，CID显示弱的RNA结合活性，但具有一定程度的序列偏好，RNA-蛋白质和肽-蛋白质界面重叠和CTD衍生的磷酸肽和RNA竞争结合位点。我们建议，蛋白质-肽和蛋白质-RNA相互作用之间的竞争是重要的机械和所需的多聚腺苷酸化因子从RNAPII脱离。
In Saccharomyces cerevisiae, the cleavage/polyadenylation factor Pcf11 is an important regulatory factor required for recruiting the polyadenylation machinery to the elongating RNA polymerase II (RNAPII) and is necessary for correct transcriptional termination. The interaction with RNAPII is mediated by a CTD-interacting domain (CID) located in the N-terminal region of Pcf11 that binds in a phospho-dependent manner the heptad repeats in the RNAPII CTD. We have previously investigated this protein-protein interaction. We examine here the interaction of the CID with different RNA sequences and look at the effect of phosphopeptides derived from the CTD heptad repeats on the RNA-protein interaction. Our findings demonstrate that the CID displays weak RNA-binding activity, but with some degree of sequence preference, the RNA-protein and peptide-protein interfaces overlap and the CTD-derived phosphopeptides and RNA compete for the binding site. We propose that competition between the protein-peptide and the protein-RNA interaction is important mechanistically and required for the disengagement of polyadenylation factors from RNAPII.