Elongation and Termination of Transcripts by RNA Pol II

描述（由申请人提供）：RNA聚合酶II合成信使RNA前体是基因表达的主要事件，对细胞生命至关重要。由每个基因产生的mRNA的量是每个相应蛋白质产生量的关键决定因素。因此，RNA pol II的功能受到细胞的极其仔细的调节。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链的起始水平或在其延伸水平进行调节。起始被认为是通过控制RNA聚合酶到基因启动子区域的募集来调节的，但是延伸是如何控制的还不太清楚。在许多临床上重要的例子中，伸长控制是重要的。HIV在延长水平调节其转录，并且未能调节c-myc癌基因转录物的延长有助于伯基特淋巴瘤的发展。在这个建议中，遗传和生物化学的方法将被用来研究pol II在动物细胞和芽殖酵母中的转录延伸和终止。转录机器的蛋白质在酵母和哺乳动物之间高度保守，并且酵母为遗传学提供了显着的实验优势。本研究的具体目的是：目的1：确定延伸因子如何影响芽殖酵母中pol Ⅱ对单个基因的转录和终止。目标二：确定pol II CTD在编码多聚腺苷酸化和非多聚腺苷酸化RNA的基因的3'末端终止转录中的作用。目标3：确定与转录pol II相关的前mRNA加工因子如何影响产生多聚腺苷酸化和非多聚腺苷酸化转录物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.