Effects of Poly(A) Binding Protein on Translational Control

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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