STRUCTURE AND FUNCTION OF EUKARYOTIC INITIATOR TRNA

真核起始子 TRNA 的结构和功能

• 批准号: 3306455
• 负责人: UTTAM L RAJBHANDARY
• 金额: $ 16.09万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-02-01 至 1996-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3306455
• 关键词: binding proteins; eukaryote; gene expression; nucleic acid sequence; protein biosynthesis; protein structure function; ribosomes; site directed mutagenesis; tissue /cell culture; transfer RNA; translation factor

The long range objective of this proposal is to understand the relationship between structure and function of eukaryotic initiator tRNAS, including the molecular mechanisms underlying their specific recognition by components of the protein synthesis machinery. Because of their unique role in initiation of protein biosynthesis, initiator tRNAS from prokaryotic and eukaryotic sources possess biochemical properties which are distinct from those of non-initiator tRNAS. These include (i) specific binding to the initiation factor eIF-2, (ii) direct binding to the P site on the ribosome and (iii) exclusion from binding to the A site on the ribosome. Along with these specific properties, initiator tRNAS possess unique sequence and/or structural features which are not found in non-initiator tRNAS. The immediate objective of this proposal is to understand the relationship between these features and the above properties of eukaryotic initiator tRNAS. The approach to be used involves isolation and analysis of properties of mutant human tRNAS. Site specific mutations will be used to generate different classes of mutant tRNAs: (a) those derived from initiator tRNAs which have lost one, two or all three of the features unique to initiator tRNAs and (b) those derived from the non-initiator methionine tRNA which now possess one, two or all three of the structural features unique to initiator tRNA. The function and detailed properties of these and other mutant tRNAs will be studied in vivo, in the yeast S. cerevisiae and in mammalian cells, and in vitro.

该提议的远距离目标是了解关系 在真核引发剂TRNA的结构和功能之间，包括 分子机制是由成分的特定识别的基础 蛋白质合成机制。因为它们在开始中的独特作用 蛋白质生物合成，原核和真核的引发剂TRNA 来源具有与生化特性不同的生化特性 非发射剂TRNA。 这些包括（i）特定的与启动的结合 因子EIF-2，（ii）直接结合核糖体上的P位点和（iii） 排除与核糖体上A位点的结合。 以及这些 特定属性，引发剂TRNA具有独特的序列和/或 在非发射剂TRNA中找不到的结构特征。 这 该提议的直接目标是了解关系 这些特征与真核引发剂的上述特性之间 trnas。 要使用的方法涉及隔离和分析 突变人类TRNA的性质。 特定站点的突变将用于 生成不同类别的突变trnas：（a） 损失了一个，两个或全部三个功能的启动器trnas 启动器trnas和（b）源自非发射因素的唯一 蛋氨酸tRNA现在拥有一个，两个或全部三个结构 启动器tRNA独有的功能。 功能和详细属性 这些和其他突变的TRNA将在酵母中进行体内研究。 酿酒酵母和哺乳动物细胞和体外。