Biophysical Studies on the Additive Effects of Modification in tRNA and rRNA

tRNA 和 rRNA 修饰相加效应的生物物理学研究

• 批准号: 0078501
• 负责人: Edward Nikonowicz
• 金额: $ 45.24万
• 依托单位: William Marsh Rice University
• 依托单位国家: 美国
• 项目类别: Continuing grant
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2005-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0078501&HistoricalAwards=false
• 关键词: Biophysical; Studies; Additive; Effects; Modification

Nikonowicz, Edward P.MCB 0078501The goal of this project is to understand key aspects of the post-transcriptional modification process of tRNA at the molecular level, including the basis of molecule specific RNA recognition by modification enzymes and the functional basis of modifications within RNA molecules. This research focuses on modification sites within the anticodon stem-loop E. coli tRNAPhe and the rRNA target of the dual specificity modification enzyme pseudouridine synthase RluA. Specifically, the following topics will be addressed: (1) chemical and enzymatic protection of MiaA RNA ligands to identify positions on the RNA that contact MiaA protein and to monitor binding induced RNA structure changes, (2) structural studies of an RNA ligand in complex with MiaA, and (3) structure determination of Helix-35 from E. coli 23S rRNA, the "second" target of the RluA enzyme. The results of these studies are directly relevant to understanding the specificity and affinity of protein-RNA interactions and the impact of modification on RNA structure and dynamics and its relationship to RNA function (e.g. translation). These investigations also will complement mechanistic studies of the prenylation reaction catalyzed by MiaA. Many RNA molecules interact with specific protein molecules either to be modified by those proteins or to perform unique cellular functions as a protein-RNA complex. Thus, it is of fundamental biological importance to learn the molecular principles that encode the specificity of individual protein-RNA interactions. At present, the molecular details of protein-RNA recognition are known for only a small number of complexes and even fewer general principles for recognition have emerged. The overall goal of this research is to determine the rules of protein-RNA recognition for two model bacterial proteins and to determine the structural and stability effects introduced by their enzymatic action.

Nikonowicz，Edward P.MCB 0078501该项目的目的是了解分子水平上TRNA的转录后修饰过程的关键方面，包括通过修饰酶和RNA分子中修饰的功能基础的分子特异性RNA识别的基础。这项研究着重于反密码子茎环大肠杆菌中的修饰位点和双重特异性修饰酶Pseudouridine合成酶RLUA的rRNA靶标。 Specifically, the following topics will be addressed: (1) chemical and enzymatic protection of MiaA RNA ligands to identify positions on the RNA that contact MiaA protein and to monitor binding induced RNA structure changes, (2) structural studies of an RNA ligand in complex with MiaA, and (3) structure determination of Helix-35 from E. coli 23S rRNA, the "second" target of the RluA enzyme.这些研究的结果与了解蛋白-RNA相互作用的特异性和亲和力直接相关，以及修饰对RNA结构和动力学及其与RNA功能的关系的影响（例如翻译）。这些研究还将补充由MIAA催化的原始反应的机理研究。许多RNA分子与特定的蛋白质分子相互作用要么由这些蛋白质修饰，要么作为蛋白RNA复合物进行独特的细胞功能。因此，学习编码单个蛋白-RNA相互作用的特异性的分子原理是基本的生物学重要性。目前，蛋白-RNA识别的分子细节仅以少量的复合物而闻名，甚至已经出现了更少的一般原则。 这项研究的总体目标是确定两种模型细菌蛋白的蛋白-RNA识别规则，并确定其酶促作用引入的结构和稳定性效应。