Elongation and Termination of Transcripts by RNA Pol II

描述(由申请人提供)：由RNA聚合酶II合成信使RNA前体是基因表达中的主要事件，对细胞的生命至关重要。每种基因产生的信使核糖核糖核酸的量是每种相应蛋白质产生多少的关键决定因素。因此，RNA PolII的功能由细胞极其谨慎地调节。这种转录过程的破坏是癌症的主要原因，干扰病毒的转录程序是治疗的潜在目标。转录可以在RNA链的起始水平或其延伸水平上进行调节。启动被认为是通过控制RNA聚合酶的招募到基因的启动子区域来调节的，但它是如何控制延伸的还不太清楚。在一些临床上重要的例子中，弹性控制很重要。HIV在转录水平上调节c-myc癌基因的转录，而c-myc癌基因转录本未能调节c-myc转录产物的伸长与Burkitt淋巴瘤的发生有关。在这项提案中，将使用遗传和生化方法来研究PolII在动物细胞和发芽酵母中的转录延伸和终止。转录机制的蛋白质在酵母和哺乳动物之间高度保守，酵母为遗传学提供了显着的实验优势。本工作的具体目的是：目标1：确定伸长因子如何影响芽殖酵母中单个基因的转录和PolII的终止。目的：研究PolII CTD在多腺苷和非多腺苷RNA编码基因3‘端转录终止中的作用。目的3：确定与转录POLII相关的前mRNA处理因子如何影响产生多腺苷和非多腺苷转录本的基因的延伸和终止。

DESCRIPTION (provided by applicant): The synthesis of messenger RNA precursors by RNA polymerase II is the primary event in gene expression and is central to the life of cells. The amount of mRNA made from each gene is a key determinant of how much of each corresponding protein is made. For this reason RNA pol II function is extremely carefully regulated by the cell. Corruption of this process of transcription is a major cause of cancer and interference with the transcription program of viruses is a potential target for therapeutics. Transcription can be regulated at the level of initiation of the RNA chains or at the level of their elongation. Initiation is thought to be regulated by controlling recruitment of RNA polymerase to a gene's promoter region but it is much less clear how elongation is controlled. Elongational control is important in a number of clinically important examples. HIV regulates its transcription at the level of elongation and failure to regulate elongation of c-myc oncogene transcripts contributes to development of Burkitt's lymphoma. In this proposal, genetic and biochemical approaches will be used to study pol II transcriptional elongation and termination in animal cells and in budding yeast. The proteins of the transcriptional machinery are highly conserved between yeast and mammals and yeast offers significant experimental advantages for genetics. The specific aims of this work are: Aim 1: To determine how elongation factors influence transcription and termination by pol II on individual genes in budding yeast. Aim 2: To determine the role of the pol II CTD in termination of transcription at the 3' ends of genes encoding polyadenylated and non-polyadenylated RNAs. Aim 3: To determine how pre-mRNA processing factors that associate with transcribing pol II affect elongation and termination at genes that produce polyadenylated and non-polyadenylated transcripts.