VPW: Mechanistic Investigation of Transcription Initiation and Elongation by E. coli RNA Polymerase (Biochemistry)

VPW：大肠杆菌 RNA 聚合酶转录起始和延伸的机制研究（生物化学）

• 批准号: 9252956
• 负责人: Lilian Hsu
• 金额: $ 10.47万
• 依托单位: University of California-Berkeley
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-01 至 1994-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9252956&HistoricalAwards=false
• 关键词: VPW; Mechanistic; Investigation; Transcription; Initiation

The proposed research will address two fundamental aspects of transcription by Escherichia coli RNA polymerase, namely, its tendency to undergo abortive initiation and its mechanism of translocation during transcription elongation. To clarify the first question, the P.I. will perform direct and quantitative comparisons, under a standardized reaction condition, of the extent and pattern of abortive initiation by a comprehensive set of E. coli promoters. The results of this determination may lead to sequence-specific correlations of abortive initiation with the initial transcribed sequence of each promoter. The second question is aimed at resolving two currently conflicting models of RNA polymerase translocation- one continuous and the other discontinuous-with the growth of the RNA chain. Experimentally, footprinting analysis with DNase I, exonuclease III, hydroxyl radical, and potassium permanganate will be performed on a contiguous set of specifically-halted elongating complexes. RNA polymerase is a central enzyme in the regulation of gene expression. Despite extensive investigation, many aspects of its catalytic mechanisms remain controversial and unclear. This work should forge a clearer picture of how translocation occurs. The project furthers VPW program objectives to provide opportunities for women to advance their careers in science or engineering through research, and to encourage other women to pursue careers in these areas through the investigator's enhanced visibility as a role model on the host campus. The proposed activities which contribute to the second objective include: teaching part of the introductory Biochemistry course to non-biochemistry science majors; mentoring and advising students; and directing undergraduate and graduate students working on research.

这项研究将解决两个基本问题。 大肠杆菌RNA聚合酶的转录方面， 也就是说，它的倾向，经历流产的启动和它的 转录延伸过程中的易位机制。 为了澄清第一个问题，P.I.将直接执行， 在标准化反应下的定量比较 条件，由一个失败的启动的程度和模式 E. coli启动子。 的结果 确定可能导致序列特异性相关性， 每个基因的初始转录序列的失败启动 启动子 第二个问题旨在解决两个 目前相互矛盾的RNA聚合酶易位模型 一个是连续的，另一个是不连续的， 的RNA链。 实验上，足迹分析与 DNA酶I、核酸外切酶III、羟基自由基和钾 高锰酸盐将在一组连续的 特异性停止的延伸复合物。 RNA聚合酶是调节细胞增殖的中心酶， 的基因表达。 尽管进行了广泛的调查，许多 其催化机制的各个方面仍然存在争议， 不清楚 这项工作应该能更清楚地说明 发生易位。 该项目进一步推动了VPW计划的目标， 妇女在科学领域发展事业的机会 或通过研究工程，并鼓励其他 妇女在这些领域从事职业， 调查员作为东道国榜样的知名度得到提高 校园 拟议的活动有助于 第二个目标包括：教学的导论部分 非生物化学专业开设生物化学课程; 指导和指导学生; 从事研究工作的本科生和研究生。