Mechanism of Specific Initiation by RNA Polymerase II

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Regulation of the frequency of transcription initiation from promoters recognized by eukaryotic RNAase II plays a major role in changing the pattern of gene expression during development and differentiation. Very little is known about the mechanism(s) of this regulation, or, indeed, about the basic mechanism of transcription initiation upon which regulation is imposed. The goal here is to dissect transcription initiation by RNA polymerase II into different mechanistic steps, to identify the proteins and DNA sequences that contribute to each step, and to test the generality of the mechanisms among various class II promoters with the hope of understanding which of these steps are targets of regulation and how is this regulation accomplished. Four protein factors have been partially purified that are required in addition to RNA polymerase II for reconstitution transcription in vivo. Their apparent roles suggest that a multi-protein complex assembles at the promoter prior to transcription initiation. This can be investigated in this system. Dr. Hawley has worked with both prokaryotic and eukaryotic RNA polymerase during her training and brings a real expertise to the field. This project takes a very high priority for funding.