Regulation of RNA Polymerase II transcription by the phosphatase Rtr1

项目总结/摘要 RNA聚合酶延伸和终止的控制尚未完全了解。在这项工作中，我们将 研究C-末端结构域磷酸酶Rtr 1和Ssu 72在RNA调节中的作用， 聚合酶II（RNAPII）延伸和终止。我们的初步研究结果表明，Rtr 1的缺失， 导致转录延伸期间RNAPII早期终止效率的整体变化。 先前的工作已经表明，Ssu 72功能的破坏导致RNAPII终止减少，从而导致 终结符通读缺陷。在这个建议中，我们将测试如何扰动每磷酸酶的影响， 转录延长和终止。此外，我们还将对Rtr 1进行酶促表征， Ssu 72底物特异性，以进一步表征其作用机制。最后，我们将确定 Rtr 1和Ssu 72之间的相互作用程度，以确定它们是否能够靶向唯一和/或重叠的 RNAPII C-终止结构域内的位点。总的来说，这些研究将揭示基本的过程， RNAPII转录控制在人类许多疾病状态中被破坏。

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.