Regulation of RNA Polymerase II transcription by the phosphatase Rtr1

项目摘要/摘要 RNA聚合酶的延长和终止的控制还不完全清楚。在这项工作中，我们将 研究C末端结构域磷酸酶Rtr1和Ssu72在RNA调控中的作用 聚合酶II(RNAPII)的延长和终止。我们的初步发现表明，Rtr1的缺失 导致RNAPII在转录延伸过程中早期终止的效率发生全局变化。 以前的工作表明，Ssu72功能的中断会导致RNAPII终止减少，从而导致 终结者通读缺陷。在这项提案中，我们将测试每种磷酸酶的扰动如何影响 转录延伸和终止。此外，我们还将对Rtr1和Rtr1进行酶学鉴定 Ssu72底物的特异性，以进一步表征其作用机制。最后，我们将确定 Rtr1和Ssu72之间的相互作用程度，以确定它们是否能够针对唯一和/或重叠 RNAPII C-终止域内的站点。总体而言，这些研究将阐明这一基本过程 RNAPII的转录控制，在人类的许多疾病状态中都被破坏。

Project Summary/Abstract The control of RNA Polymerase elongation and termination is not fully understood. In this work, we will investigate the role of the C-terminal domain phosphatases Rtr1 and Ssu72 in the regulation of RNA Polymerase II (RNAPII) elongation and termination. Our preliminary findings suggest that deletion of Rtr1 results in global changes in the efficiency of early termination of RNAPII during transcription elongation. Previous work has shown that disruption of Ssu72 function leads to decreased RNAPII termination resulting in terminator read-through defects. In this proposal, we will test how perturbations of each phosphatase effect transcription elongation and termination. Additionally, we will perform enzymatic characterization of Rtr1 and Ssu72 substrate specificity to further characterize their mechanisms of action. Finally, we will determine the degree of interplay between Rtr1 and Ssu72 to determine if they are able to target unique and/or overlapping sites within the RNAPII C-termination domain. Overall, these studies will shed light on the fundamental process of RNAPII transcription control which is disrupted in numerous disease states in humans.