CAREER: Exploring Conservation in the Molecular Determinants of the Fidelity Mechanism in Translation

职业：探索翻译保真机制分子决定因素的守恒性

• 批准号: 0747230
• 负责人: Scott Blanchard
• 金额: $ 65万
• 依托单位: Joan and Sanford I. Weill Medical College of Cornell University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-15 至 2013-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0747230&HistoricalAwards=false
• 关键词: CAREER; Exploring; Conservation; Molecular; Determinants

This integrated research and educational plan focuses on the application and development of single-molecule Fluorescence Resonance Energy Transfer FRET(smFRET) methods to investigate the role and function of dynamic process of protein synthesis catalyzed by the ribosome. Particular focus will be given to delineating the structural and kinetic parameters of nanometer-scale remodeling events within the ribosome during tRNA selection and translocation processes on ribosomes from different species, and how these stochastic events contribute to the mechanism of fidelity. Our principal aim is to determine conserved and divergent features of these mechanisms across evolution. The application of smFRET imaging methods, in combination with genetic, biochemical, and computation efforts, will provide quantitative measures of the ribosome energy landscape at high-spatial and -time resolution. Towards this goal, advances will be made in the construction of biologically compatible microfluidic systems, the design of computational tools required for the analysis of single-molecule data, and in the stabilization of fluorescent dye molecules. The successful integration and completion of this project is expected to advance the understanding of conserved mechanisms in protein synthesis across evolution. A deeper knowledge of the role of dynamic structural processes within the ribosome will have a broad impact, spanning any enzymological system where conformational changes impact function and regulation in the cell. The nature of this research provides a unique training environment for young scientists where new technologies and cross-disciplinary expertise is required to engage effectively their scientific pursuits.

该综合研究和教育计划侧重于单分子荧光共振能量转移FRET（smFRET）方法的应用和发展，以研究核糖体催化蛋白质合成动态过程的作用和功能。特别关注的将是描绘的结构和动力学参数的核糖体内的tRNA选择和易位过程中的核糖体从不同的物种，以及如何这些随机事件有助于保真度的机制。我们的主要目的是确定这些机制在进化过程中的保守和不同特征。smFRET成像方法的应用，结合遗传，生物化学和计算的努力，将提供定量措施的核糖体能量景观在高空间和时间分辨率。为了实现这一目标，将在生物相容的微流体系统的建设，设计所需的计算工具的单分子数据的分析，并在荧光染料分子的稳定化方面取得进展。该项目的成功整合和完成有望促进对蛋白质合成中保守机制的理解。深入了解核糖体内动态结构过程的作用将产生广泛的影响，跨越任何酶系统，其中构象变化影响细胞中的功能和调节。这项研究的性质为年轻科学家提供了一个独特的培训环境，需要新技术和跨学科专业知识来有效地从事他们的科学追求。