FUNCTIONAL ANALYSIS OF AN RNA STRUCTURAL MOTIF

RNA 结构基序的功能分析

• 批准号: 6077089
• 负责人: Ya-Ming Hou
• 金额: $ 2.61万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-17 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6077089
• 关键词: Archaea; Escherichia coli; SDS polyacrylamide gel electrophoresis; X ray crystallography; acylation; aminoacyl tRNA; bacterial RNA; cystathionine beta synthase; cysteine; enzyme activity; high performance liquid chromatography; molecular cloning; mutagen testing; nuclear magnetic resonance spectroscopy; nucleic acid sequence; nucleic acid structure; physical model; polymerase chain reaction; protein structure function; site directed mutagenesis; structural biology; sugar phosphates; transfer RNA; uracil nucleotides

DESCRIPTION: (Adapted from Applicant's Abstract) Although various aspects of transfer RNA (tRNA) recognition by aminoacryl-tRNA synthetases have been well established, that of structural recognition is not understood. The study of recognition of E. coli tRNACys by cysteine-tRNA synthetase offers the potential that questions of tRNA structural recognition can be addressed. One objective is to determine the mechanism of contribution of a single nucleotide U73 to aminoacylation. A second objective is to investigate the scope and impact of a structural recognition of an unusual G15:G48 base pair that is unique to E. coli tRNACys. A third objective is to study the potentially novel mechanism of aminoacylation with cysteine in an archaebacterium. In aim 1, the hypothesis that U73 uses its functional groups to contribute to aminoacylation will be tested. In aim 2, the hypothesis that U73 uses its ability to confer a fold-back structure of the CCA end to contribute to aminoacylation will be tested. In aim 3, the biologically relevant structure of the important G15:G48 in E. coli tRNACys will be defined and tested, In aim 4, the hypothesis that G15:G48 coordinates with multiple nucleotides to contribute to aminoacylation will be tested. In aim 5, the hypothesis that tRNA structural recognition is the primary difference between the E. coli and human systems of aminoacylation with cysteine will be tested. In aim 6, the hypothesis that aminoacylation of tRNACys in the archaebacterium Halobacter volcanii (H. volcanni) is catalyzed by a novel cysteine-tRNA synthetase will be tested. The results of these studies will be integrated into structural studies of tRNACys and its complex with cysteine-tRNA synthetase in order to have a better understanding of tRNA structural recognition by an aminoacyl-tRNA synthetase.

描述：（根据申请人的摘要改编），尽管 通过氨基丙烯酰基-TRNA识别转移RNA（tRNA）的各个方面 合成酶已经建立了结构识别 不了解。 研究的研究 半胱氨酸-TRNA合成酶的潜力是tRNA的问题 可以解决结构识别。一个目的是确定 单个核苷酸U73对 氨基酰化。 第二个目标是调查范围和 对不寻常的G15：G48碱基对的结构识别的影响 大肠杆菌trnacys是独特的。 第三个目标是研究 与半胱氨酸的氨基酰化的潜在新型机制 考古细菌。 在AIM 1中，U73使用其官能团的假设 将测试有助于氨基酰化。 在AIM 2中，假设 U73使用其赋予CCA末端的折叠结构的能力 为氨基化作用做出贡献。 在AIM 3中 大肠杆菌中重要的G15：G48的生物学相关结构 在AIM 4中，将定义和测试Trnacys。 G15：G48与多个核苷酸坐标有助于 氨基酰化将进行测试。 在AIM 5中，tRNA的假设 结构识别是大肠杆菌和大肠杆菌之间的主要区别 用半胱氨酸的氨基酰化系统将进行测试。 目标 6，假设trnacys在考古细菌中的氨基酰化 Halobacter Volcanii（H. volcanni）被一种新颖的半胱氨酸-TRNA催化 合成酶将进行测试。 这些研究的结果将是 与Trnacys的结构研究及其复合 半胱氨酸-tRNA合成酶为了更好地了解tRNA 氨基酰基-TRNA合成酶的结构识别。