Elongation and Termination of Transcripts by RNA Pol II

RNA Pol II 转录本的延伸和终止

• 批准号: 7085486
• 负责人: DAVID L BENTLEY
• 金额: $ 22.63万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2008-03-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7085486
• 关键词: DNA directed RNA polymerase; RNA biosynthesis; RNA splicing; Xenopus; Xenopus oocyte; enzyme activity; fungal genetics; genetic regulatory element; genetic transcription; immunoprecipitation; messenger RNA; microarray technology; nucleic acid hybridization; phosphorylation; polyadenylate; polymerase chain reaction; transcription factor; transcription termination; yeasts

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.

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，对细胞的生命至关重要。每个基因产生的mRNA的数量是每个相应蛋白质产生多少的关键决定因素。因此，RNA pol II的功能受到细胞的严格调控。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。起始被认为是通过控制RNA聚合酶在基因启动子区域的招募来调节的，但如何控制延伸则不太清楚。在一些临床重要的例子中，伸长控制是很重要的。HIV在伸长水平上调控其转录，未能调控c-myc癌基因转录物的伸长有助于伯基特淋巴瘤的发展。在这个提议中，遗传和生化的方法将被用于研究动物细胞和出芽酵母中pol II转录的延伸和终止。酵母和哺乳动物之间的转录机制蛋白高度保守，酵母为遗传学提供了显著的实验优势。这项工作的具体目的是：目的1：确定伸长因子如何影响芽殖酵母中pol II对单个基因的转录和终止。目的2：确定pol II CTD在编码多聚腺苷化和非多聚腺苷化rna的基因3'端转录终止中的作用。目的3：确定与转录pol II相关的mrna前加工因子如何影响产生聚腺苷化和非聚腺苷化转录本的基因的延伸和终止。