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Mechanism of RNA-dependent kinase PKR expression. In a relentless attempt to meet the project's goals in their entirety, it is only a minor drawback that the challenging time frame did not suffice for the assignment's flawless completion. The murine RNA-dependent protein kinase, which derives its significance from the fact that it is a key mediator in a cell's antiviral activity was the focus of our study. As a response to a viral attack, PKR autophosphorylates itself and then phosphorylates other proteins in a sequence of events that triggers the eventual "shut down" of the cell. Our research goal for the summer was to successfully isolate PKR from E.Coli cells and to determine several of its biophysical and biochemical characteristics, as well as its complexes formed with target substrate proteins and activating double-stranded RNAs. This investigation of its structure might help us to clear up many urgent questions considering the mechanistic properties of PKR. In order to do this, we were going to head towards the production of crystals in order to determine its exact structure via Xray crystallography. Eventually, the detailed structural analysis of PKR could lead to a variety of experiments to synthesize small-molecule ligands which are able to jeopardize TAR RNA and Tat protein functions (both HIV components) that disrupt the antiviral activity of PKR. Unfortunately, we had difficulties with the purification of PKR from E.Coli cells. Return to the RISE 2000 project list |
