Characterization of translation elongation mechanisms

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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.