Effects of Poly(A) Binding Protein on Translational Control

0076344Goss该项目旨在确定多聚（A）结合蛋白（PABP）刺激蛋白质合成的刺激机制，并将解旋酶相互作用与翻译效率相关联。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旋活性。通过将解旋速率与翻译速率相关联，可以分析解旋酶反应所需的程度以及解旋酶反应是否限制速率。通过检查定量结合和动力学，将获得洞察力的二级结构，阻止翻译结果从不利的结合平衡或抑制限速步骤。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。该项目的实验是第一个针对解旋酶活性和翻译效率之间的详细联系。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节的过程，使细胞能够快速响应环境的变化，并节省材料和能源。 蛋白质合成是有效生长和发育所必需的。 虽然有大量的信息是必要的组成部分和蛋白质合成的一般途径，知道的速率限制步骤和这些过程的分子细节少得多。 为了调节一个过程，人们希望影响限速步骤，并确定正常和异常蛋白质合成之间的关键差异。 更好地理解蛋白质合成的分子机制将导致疾病状态的可能治疗目标，增加生长效率，可能使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