INVESTIGATION OF THE BINDING SITE OF RNA POLYMERASE ON DKSA

DKSA 上 RNA 聚合酶结合位点的研究

• 批准号: 7954670
• 负责人: Richard L. Gourse
• 金额: $ 0.02万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 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/7954670
• 关键词: Amino Acids; Binding Sites; Complex; Computer Retrieval of Information on Scientific Projects Database; DNA-Directed RNA Polymerase; Escherichia coli; Funding; Genetic Transcription; Grant; Half-Life; Homologous Gene; Institution; Investigation; NMR Spectroscopy; Nuclear Magnetic Resonance; Research; Research Personnel; Resources; Sequence Analysis; Source; Structure; Testing; Time; United States National Institutes of Health; promoter; thermophilic organism

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. At this time, all x-ray structures for holo and core RNAP are from the thermophiles T. thermophilus and T. aquaticus, neither of which appears to encode a DksA homolog recognizable from sequence analysis. We tested whether E. coli DksA would function on T. thermophilus or T. aquaticus RNAP. Unfortunately, E. coli DksA neither inhibited transcription nor reduced the half-lives of promoter complexes formed by these enzymes.Therefore, we concluded that it would not be prudent to attempt to get crystals for a heterologous complex containing E. coli DksA and T. thermophilus RNAP. Therefore, we have decided to use nuclear magnetic resonance (NMR) spectroscopy to define amino acid residues in E. coli DksA that are contacted by E. coli RNAP.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 此时，全RNAP和核RNAP的所有X射线结构都来自嗜热菌T。thermophilus和T. aquaticus，这两个似乎都没有编码可从序列分析中识别的DksA同源物。我们测试了E. coliDksA在T. thermophilus或T.水生植物RNAP。不幸的是，E. coli DksA既不抑制转录，也不减少由这些酶形成的启动子复合物的半衰期。coli DksA和T.嗜热菌RNAP因此，我们决定使用核磁共振（NMR）光谱来定义E中的氨基酸残基。coliDksA与E. coli RNAP。