Effects of Poly(A) Binding Protein on Translational Control

本项目旨在确定Poly (A) Binding Protein （PABP）刺激蛋白合成的机制，并将解旋酶相互作用与翻译效率联系起来。结合和解旋酶活性的直接荧光测定将用于确定这些因子的解绕活性。通过将解绕速率与翻译速率相关联，可以分析解旋酶反应需要的程度，以及解旋酶反应是否具有速率限制。通过研究定量结合和动力学，我们将了解到阻碍翻译的二级结构是由不利的结合平衡还是限速步骤的抑制引起的。最终，翻译是一个高度调控的过程，不仅具有严格控制的速率，而且还决定了mRNA的选择。这个项目的实验是第一个在解旋酶活性和翻译效率之间建立详细联系的实验。该项目将使用已经了解的生化分析以及新的生物物理分析，这将导致这些过程的详细动力学分析。蛋白质合成是一个高度调控的过程，它使细胞能够快速响应环境的变化，并且节省物质和能源资源。蛋白质的合成是有效生长发育所必需的。虽然关于蛋白质合成的必要成分和一般途径的信息很多，但对这些过程的限速步骤和分子细节所知甚少。为了调节一个过程，人们想要影响限速步骤，并确定正常和异常蛋白质合成之间的关键区别。更好地了解蛋白质合成的分子机制将导致疾病状态的可能治疗靶点，提高生长效率，可能利用植物生产疫苗和抗病毒策略

0076344Goss This project seeks to determine the mechanism of stimulation of Poly (A) Binding Protein (PABP) stimulation of protein synthesis and to correlate the helicase interaction with translational efficiency. A direct fluorescence assay for binding and helicase activity will be utilized to determine the unwinding activity of these factors. By correlating the rate of unwinding with the rate of translation, an analysis can be made of the extent to which the helicase reaction is required and if it is rate limiting. By examining quantitative binding and kinetics, insight will be gained on whether secondary structure that blocks translation results from unfavorable binding equilibria or inhibition of rate-limiting steps. Ultimately, translation is a highly regulated process that not only has a closely controlled rate, but also determines the selection of mRNA. The experiments of this project are among the first directed at making detailed connections between helicase activity and translational efficiency. This project will use biochemical assays already understood as well as new biophysical assays that will lead to detailed kinetic analyses of these processes. Protein synthesis is a highly regulated process that allows a cell to respond to changes in the environment rapidly and with and economy of material and energy resources. Protein synthesis is necessary for efficient growth and development. While a great deal of information is available about the components necessary and the general pathways for protein synthesis, much less is known about the rate-limiting steps and molecular details of these processes. In order to regulate a process, one wants to affect the rate-limiting step and identify the crucial differences between normal and abnormal protein synthesis. A better understanding of the molecular mechanism of protein synthesis will lead to possible therapeutic targets for disease states, increased growth efficiency, possible uses of plants for production of vaccines, and anti-viral strategies