Characterization of the Interactions of Eukaryotic Initiation Factors, Ribosomes, and mRNA

真核起始因子、核糖体和 mRNA 相互作用的表征

• 批准号: 9722907
• 负责人: Dixie Goss
• 金额: $ 34.13万
• 依托单位: CUNY Hunter College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-10-01 至 2000-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9722907&HistoricalAwards=false
• 关键词: Characterization; Interactions; Eukaryotic; Initiation; Factors

9722907 Goss The long term goal of this project is to understand the mechanism of eukaryotic protein synthesis. Eukaryotes initiate translation differently in normal cells than in virus infected cells or cells under stress conditions such as heat shock. The overall scheme of protein synthesis has been described, but the details and particularly the kinetics of this process remain to be elucidated. The equilibrium and kinetics of the interaction of eukaryotic initiation factors (eIFs) with mRNA and oligonucleotides will be examined. The effects of eIFs on ribosome binding to several oligonucleotides and mRNA will be determined. Direct fluorescence measurements will be used to elucidate the interaction of initiation factors, ribosomes and mRNA or oligonucleotides. These multiple interactions will be analyzed to determine if the binding interactions are cooperative, anti-cooperative, or independent. Kinetic results will determine the rate limiting steps and the probable pathway of interactions. Quantitative data will give insight into the effect of mRNA secondary structure on ribosome binding, the possible communication between the 5' and 3' termini of RNA, and the possibility of internal ribosome entry in higher plant systems. These results will aid in formulating a detailed molecular mechanism for the assembly of the eukaryotic initiation complex. This project will elucidate the mechanism for protein synthesis in plant systems. Regulation of protein synthesis is important in regulation of cell growth and development as well as in biotechnology applications. Plants provide possible hosts for production of vaccines and other proteins of commercial interest without the contaminants that may be present from a bacterial or other host used for gene cloning. It is therefore important to understand the regulation of protein synthesis in plants. These studies will determine the rate limiting steps for interactions that determine the production of proteins. By elucidating these steps o ne can then determine the possible intervention points to increase or decrease the rate of the overall protein synthesis process.

9722907高斯这个项目的长期目标是了解真核生物蛋白质合成的机制。 真核细胞在正常细胞中启动翻译的方式与病毒感染细胞或热休克等应激条件下的细胞不同。蛋白质合成的总体方案已经描述，但细节，特别是这个过程的动力学仍有待阐明。 将检查真核起始因子（eIFs）与mRNA和寡核苷酸相互作用的平衡和动力学。 将确定eIF对核糖体与几种寡核苷酸和mRNA结合的影响。 直接荧光测量将用于阐明起始因子、核糖体和mRNA或寡核苷酸的相互作用。 将分析这些多重相互作用以确定结合相互作用是协同的、反协同的还是独立的。 动力学结果将确定速率限制步骤和相互作用的可能途径。 定量数据将使深入了解mRNA二级结构对核糖体结合的影响，RNA的5'和3'末端之间可能的通讯，以及高等植物系统中内部核糖体进入的可能性。 这些结果将有助于制定一个详细的真核起始复合物的组装分子机制。 本项目将阐明植物蛋白质合成的机制 系统. 蛋白质合成的调节在细胞生长和发育的调节以及生物技术应用中是重要的。 植物为生产疫苗和其他具有商业价值的蛋白质提供了可能的宿主，而没有可能存在于用于基因克隆的细菌或其他宿主中的污染物。 因此，了解植物蛋白质合成的调控是很重要的。 这些研究将确定决定蛋白质产生的相互作用的限速步骤。通过阐明这些步骤，然后可以确定可能的干预点，以增加或减少整个蛋白质合成过程的速率。