RNA PROTEIN INTERACTIONS IN E COLI RHO

大肠杆菌 RHO 中的 RNA 蛋白质相互作用

• 批准号: 6012984
• 负责人: T Kevin HITCHENS
• 金额: $ 3.67万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6012984
• 关键词: Escherichia coli; RNA binding protein; bacterial DNA; bacterial RNA; bacterial proteins; intermolecular interaction; model design /development; nuclear magnetic resonance spectroscopy; physical model; protein structure function; structural biology; thermodynamics; transcription termination

This proposal focuses on the investigation of structure, and specific protein-nucleotide interactions of the RNA-binding domain of E. coli. rho protein. This protein plays an important role in the termination of transcription in prokaryotic organisms. Rho is particularly intriguing in that it binds equally well single- stranded DNA and single-stranded DNA however, it is only activated when bound to RNA. NMR spectroscopy methods will be used to determine the structures of-the rho RNA binding domain when bound to single stranded RNA and single stranded DNA. Various oligonucleotide ligands and protein mutations will be used to probe the specific interactions between the protein and nucleic acids to further the understand molecular mechanism of rho-dependent transcription termination. While much has been learned about DNA binding protein in recent years, relatively little is known about RNA binding proteins. Characterization of the binding interactions of rho with nucleic acids will be helpful in understanding RNA binding in other proteins since the RNA binding domain has structural homology with many prokaryotic and eukaryotic RNA-binding proteins.

本研究的重点是研究大肠杆菌RNA结合结构域的结构和特异性蛋白质-核苷酸相互作用。杆菌rho蛋白。这种蛋白质在原核生物中的转录终止中起重要作用。Rho是特别有趣的，因为它同样很好地结合单链DNA和单链DNA，然而，它仅在结合RNA时被激活。NMR光谱法将用于确定rho RNA结合结构域与单链RNA和单链DNA结合时的结构。我们将利用各种寡核苷酸配体和蛋白质突变来探测蛋白质和核酸之间的特异性相互作用，以进一步了解rho依赖性转录终止的分子机制。虽然近年来对DNA结合蛋白的了解很多，但对RNA结合蛋白的了解相对较少。rho与核酸的结合相互作用的表征将有助于理解其他蛋白质中的RNA结合，因为RNA结合结构域与许多原核和真核RNA结合蛋白具有结构同源性。