STRUCTURE AND FUNCTION OF YEAST TRANSLATION FACTOR EIF3

酵母翻译因子 EIF3 的结构和功能

• 批准号: 6635788
• 负责人: DAVID V FREISTROFFER
• 金额: $ 1.34万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-01 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6635788
• 关键词: binding sites; crosslink; fungal genetics; messenger RNA; microarray technology; polymerase chain reaction; protein structure function; translation factor; yeasts

Gene expression in eukaryotes is often controlled at the level of the initiation of translation. eIF3 plays a central role in this process. It is responsible for ribosome subunit dissociation, it stabilizes the initiator Met-tRNA on the small ribosomal subunit and it is required for mRNA binding to the ribosome. p33, an RNA-binding subunit of eIF3, has recently been cloned and partially characterized. p33 is part of the "core" of the eIF3 complex and is an essential protein in yeast. The elucidation of the function of the p33 subunit, the function of eIF3, and the mechanism of translation initiation in yeast in a longer perspective will be sought in the research under this training period via these four specific aims: 1. Identify the physiological RNA target for p33 binding using SELEX. 2. Isolate high and low copy number extragenic suppressors of p33 mutants. 3. Use microarrays and kinetic RT-PCR to examine mRNA from polysome fractions in p33 mutants. . 4. Elucidate the structure/function of eIF3 by protein crosslinking and by forming partial complexes (e.g., p33, p39 and p93) and testing their activities in vitro.

真核生物中的基因表达通常被控制在翻译起始的水平上。EIF3在这一过程中发挥着核心作用。它负责核糖体亚基的解离，稳定小核糖体亚基上的启动子Met-tRNA，是mRNA与核糖体结合所必需的。P33是eIF3的一个RNA结合亚基，最近被克隆并部分鉴定。P33是eIF3复合体“核心”的一部分，是酵母中必不可少的蛋白质。在本次培训期间的研究中，将寻求从更长远的角度阐明p33亚基的功能、eIF3的功能以及酵母中翻译启动的机制：1.利用SELEX鉴定p33结合的生理RNA靶点。2.分离高拷贝数和低拷贝数的p33突变体基因外抑制物。3.应用基因芯片和动态RT-PCR技术检测p33突变体中多聚体部分的mRNA。。4.通过蛋白质交联和形成部分复合体(如p33、p39和p93)并在体外检测它们的活性，阐明eIF3的结构和功能。