Characterization of translation elongation mechanisms

基因表达是将DNA解码为RNA转化为蛋白质的基本过程。将RNA解码成蛋白质，称为翻译，涉及一个名为核糖体的大型复杂大分子机器。核糖体读取并沿着RNA序列移动，从而将氨基酸顺序添加到新生蛋白质中。显然，翻译是受高度调控的，翻译的速度和保真度会在不同的生物情况下受到影响，如细胞应激和病毒感染。目前，影响核糖体解码和运动的确切调控和机制还知之甚少。由于其根本重要性，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，我们开发了一种新的最小重组翻译系统，以描述所涉及的机制和因素。这个系统将使我们能够系统地一次添加一个因素，以阐明每一步的功能。此外，这个系统提供了一个独特的机会来研究起始下游的事件，如RNA的运动和解码。使用生化方法，我们将描述核糖体如何准确地解码RNA，以及各种因素如何调节核糖体的运动。最后，由于RNA可以折叠并形成稳定的结构，我们还将测试核糖体如何相互作用和解开这些RNA结构。通过利用这个最小的系统，这些实验将增强我们对基本基因表达机制的知识。

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, 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 the movement of the ribosome. Finally, 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.