RNA-binding RNA Polymerase II CTD-Associated Proteins

描述（由申请人提供）： 项目摘要。这项建议的主要目的是了解RNA聚合酶II终止多聚腺苷酸化和非多聚腺苷酸化转录本的机制。控制这一过程的因素和条件在决定哪些基因表达及其表达水平方面起着重要作用。我们将继续研究酿酒酵母RNA结合蛋白Nrd1，Nab3和RNA解旋酶Sen1在转录终止中的作用。我们还将研究RNA聚合酶II CTD磷酸化的作用和一些其他RNA结合蛋白的功能，这些蛋白与多聚腺苷酸化和非多聚腺苷酸化转录物的终止有关。我们将使用一种新开发的交联技术来绘制RNA聚合酶II及其终止因子与活细胞中新生RNA的结合。这些图谱和随后的遗传和生物化学实验将用于：（i）鉴定对不同类型的细胞起作用的终止子序列。 RNA聚合酶II转录本和（ii）确定从模板中去除RNA聚合酶II所涉及的机制。

DESCRIPTION (provided by applicant): Project Summary. The broad objective of this proposal is to understand the mechanism of RNA polymerase II termination of both polyadenylated and non-polyadenylated transcripts. The factors and conditions that control this process play important roles in determining which genes are expressed and the level of their expression. We will continue to study the roles of Saccharomyces cerevisiae RNA-binding proteins Nrd1, Nab3 and the RNA helicase Sen1 in transcription termination. We will also investigate the roles of RNA polymerase II CTD phosphorylation and the function of a number of other RNA-binding proteins implicated in termination of both polyadenylated and non-polyadenylated transcripts. We will use a newly developed cross-linking technique to map the binding of RNA polymerase II and its termination factors to nascent RNAs in living cells. These maps and subsequent genetic and biochemical experiments will be used to: (i) identify terminator sequences that operate on different classes of RNA polymerase II transcripts and (ii) determine the mechanisms involved in removing RNA polymerase II from the template.