Molecular analysis of transcription termination by human RNA polymerase II

描述（由申请人提供）：RNA聚合酶II终止的分子机制仍然知之甚少。mRNA 3'末端加工与终止偶联，并且该加工步骤的失调导致人类疾病。本项目的长期目标是表征共转录加工因子在RNA 3'末端加工和pol II转录终止中的作用。本研究的目的是：1.确定哺乳动物poly（A）位点依赖性pol Ⅱ终止的相关因子; 2.研究延伸因子Spt 5在pol Ⅱ 3'末端加工和终止中的作用。我们将进行全基因组RNAi筛选，旨在鉴定人类细胞中pol II终止所需的新因子。该测定的变化可能进一步用于鉴定在某些癌症中观察到的参与替代性聚（A）位点选择的因素。 公共卫生相关性：越来越多的人类疾病，包括癌症，其特征在于基因转录物的不正确或交替的3'端形成。mRNA 3'末端加工与RNA聚合酶终止偶联。通过这个项目，我希望确定转录终止的分子机制，从而扩大我们对这些疾病的理解。

DESCRIPTION (provided by applicant): The molecular mechanisms of RNA polymerase II termination remain poorly understood. mRNA 3' end processing is coupled to termination, and deregulation of this processing step results in human disease. The long-term objective of this project is to characterize the role of co-transcriptional processing factors in RNA 3' end processing and pol II transcriptional termination. The aims of this proposal are 1: to determine factors involved in mammalian poly(A) site dependent pol II termination, and 2: to investigate the role of the elongation factor Spt5 on pol II 3' end processing and termination. We will conduct a genome wide RNAi screen designed to identify new factors required for pol II termination in human cells. Variations on this assay might further be used to identify factors involved in alternative poly(A) site choice observed in certain cancers. PUBLIC HEALTH RELEVANCE: There are a growing number of human diseases, including cancer, that are characterized by incorrect or alternate 3' end formation of gene transcripts. mRNA 3' end processing is coupled to RNA polymerase termination. With this project I hope to determine the molecular mechanisms of transcriptional termination, and thereby expand upon our understanding of these diseases.