Molecular analysis of transcription termination by human RNA polymerase II

描述(申请人提供)：RNA聚合酶II终止的分子机制仍然知之甚少。信使核糖核酸3‘末端的处理与终止有关，而这一处理步骤的解除调控会导致人类疾病。该项目的长期目标是表征共转录加工因子在RNA3‘末端加工和PolII转录终止中的作用。本研究的目的是：1：确定哺乳动物PolII依赖PolyA位点终止的相关因素；2：研究延长因子Spt5在POLII 3‘末端加工和终止中的作用。我们将进行全基因组RNAi筛选，以确定在人类细胞中终止PolII所需的新因素。该检测的变异可能进一步用于确定某些癌症中观察到的替代Poly(A)位点选择所涉及的因素。 与公共卫生相关：包括癌症在内的越来越多的人类疾病的特点是基因转录本的3‘端不正确或交替形成。RNA3‘末端处理与RNA聚合酶终止有关。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.