IDENTIFYING THE PATH OF THE RNA PRIMER AROUND A SIMPLE RNA POLYMERASE

识别简单 RNA 聚合酶周围 RNA 引物的路径

• 批准号: 7954492
• 负责人: CHANGGONG LI
• 金额: $ 0.33万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7954492
• 关键词: Binding Sites; Complex; Computer Retrieval of Information on Scientific Projects Database; DNA-Directed RNA Polymerase; Ensure; Funding; Grant; Hour; Individual; Institution; Polymerase; Polynucleotide Adenylyltransferase; Proteins; RNA; RNA Polymerase I; RNA primers; Research; Research Personnel; Resources; Source; Structure; Surface; Tail; Time; United States National Institutes of Health; Vaccinia virus; Work; flexibility; structural biology; synchrotron radiation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Vaccinia virus poly(A) polymerase (VP55) is the only RNA polymerase known to translocate with respect to its RNA primer, raising a paradox: Which is more flexible: The polymerase or the primer? The protein has two known modes of action, namely the processive synthesis of 30 nt tails and, as a heterodimer with its processivity factor (VP39), the semi-processive synthesis of 200 ? 300 nt tails. Past, collaborative studies, have yielded crystal structures for VP39, and for the VP55-VP39 heterodimer. The most important, yet absent information is a structural understanding of the heterodimer-primer complex. By using the beam time of a colleague, a full-time crystallographer in the Gershon lab has progressed markedly in finding binding sites for short RNA segments on the polymerase surface. Although this is ground-breaking, original work for a poly(A) polymerase, 3 hours of beam time every 2 months provides inadequate progress in this NIH-funded work to ensure grant renewal. Beam time is the major bottleneck, hence the current request for individual, regular beam time for this crystallographic work.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 痘苗病毒多聚腺苷酸聚合酶 (VP55) 是唯一已知可相对于其 RNA 引物易位的 RNA 聚合酶，这引发了一个悖论：聚合酶和引物哪个更灵活？该蛋白质有两种已知的作用模式，即 30 nt 尾部的持续合成，以及作为具有持续因子 (VP39) 的异二聚体，以及 200 nt 尾部的半持续合成。 300 nt 尾巴。过去的合作研究已经获得了 VP39 和 VP55-VP39 异二聚体的晶体结构。最重要但缺乏的信息是对异二聚体-引物复合物的结构理解。通过利用同事的光束时间，Gershon 实验室的一名全职晶体学家在寻找聚合酶表面短 RNA 片段的结合位点方面取得了显着进展。尽管这是 Poly(A) 聚合酶的开创性原创工作，但每 2 个月 3 小时的光束时间在这项 NIH 资助的工作中并没有取得足够的进展，无法确保资助续签。光束时间是主要瓶颈，因此目前这项晶体学工作需要单独、定期的光束时间。