Characterization of translation elongation mechanisms

翻译延伸机制的表征

• 批准号: 341459-2012
• 负责人: Jan, Eric
• 金额: $ 2.48万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2013
• 资助国家: 加拿大
• 起止时间: 2013-01-01 至 2014-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=527315
• 关键词: Characterization; translation; elongation; mechanisms

Gene expression is the fundamental process of decoding DNA to RNA into protein. Decoding RNA into protein, called translation, involves a large complex macromolecular machine called the ribosome. The ribosome reads and moves along the RNA sequence to sequentially add amino acids to the nascent protein. It is apparent that translation is highly regulated and the rates and fidelity of translation can be influenced under different biological situations such as cellular stress and viral infection. Currently, the exact regulations and mechanisms affecting ribosome decoding and movement of the ribosome is poorly understood. Because of its fundamental importance in gene expression and regulation, it is imperative that we understand the full repertoire of translation mechanisms and regulations. To directly address these issues, we have developed a novel minimal reconstituted translation system in order to delineate the mechanisms and factors involved. This system will allow us to systematically add back factors one at a time in order to elucidate the function of each step. Moreover, this system allows the unique opportunity to study events downstream of initiation such as the movement and decoding of RNA. Using biochemical approaches, we will characterize how the ribosome accurately decodes RNA as well as how factors are mediating th movement of the ribosome. Moreover, as RNA can fold and form stable structures, we will also test how the ribosome interacts and unwinds these RNA structures. By taking advantage of this minimal system, these experiments will enhance our knowledge of basic gene expression mechanisms.

基因表达是将DNA解码为RNA为蛋白质的基本过程。将RNA解码为蛋白质，称为翻译，涉及一种称为核糖体的大型复合物分子机。核糖体沿RNA序列读取并移动，以依次将氨基酸添加到新生的蛋白质中。显然，翻译受到高度调节，在不同的生物学情况（例如细胞应激和病毒感染）下，翻译的速度和忠诚度可能受到影响。当前，核糖体的核糖体解码和运动的确切法规和机制知之甚少。由于其在基因表达和调节中的基本重要性，因此我们必须了解翻译机制和法规的全部内容。为了直接解决这些问题，我们开发了一种新颖的最小重组翻译系统，以描绘涉及的机制和因素。该系统将使我们一次系统地添加一个因素，以阐明每个步骤的功能。此外，该系统允许独特的机会研究起始下游的事件，例如RNA的运动和解码。使用生化方法，我们将表征核糖体如何准确解码RNA以及如何介导核糖体运动的因素。 此外，由于RNA可以折叠并形成稳定的结构，因此我们还将测试核糖体如何相互作用并放松这些RNA结构。通过利用这个最小系统，这些实验将增强我们对基本基因表达机制的了解。