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.

该提案的长期目标是了解两者之间的关系 真核起始子 tRNAS 的结构和功能之间的关系，包括 其成分特异性识别的分子机制 蛋白质合成机器。由于它们在启动中的独特作用 蛋白质生物合成，原核和真核生物的起始tRNAS 来源具有与其他来源不同的生化特性 非起始 tRNA。 这些包括 (i) 与起始物的特异性结合 因子 eIF-2，(ii) 直接结合至核糖体上的 P 位点，以及 (iii) 排除与核糖体 A 位点的结合。 连同这些 特定的性质，起始子 tRNAS 拥有独特的序列和/或 非起始 tRNA 中未发现的结构特征。 这 该提案的直接目标是了解这种关系 这些特征与真核引发剂的上述特性之间 tRNA。 所使用的方法包括隔离和分析 突变型人类 tRNA 的特性。 位点特异性突变将用于 产生不同类别的突变 tRNA：(a) 源自 起始 tRNA 丢失了一项、两项或全部三个特征 启动子 tRNA 所特有的，(b) 源自非启动子的 tRNA 蛋氨酸 tRNA，现在具有一种、两种或全部三种结构 具有启动子 tRNA 独有的特征。 的功能和详细属性 这些和其他突变 tRNA 将在酵母 S. 体内进行研究。 酿酒酵母和哺乳动物细胞以及体外。