This research proposal focuses on the molecular details of how the E. coli rho protein determines the location for the dissociation of the ternary transcription complex at specific sites on the DNA template. A model has been previously suggested that postulates that the interaction of rho protein with large unstructured regions of the transcript activates the ATPase-dependent rho-catalyzed transcript release reaction. RNA polymerase pause sites kinetically determine the position on the template where the release reaction is most likely to take place. This model will be used to design synthetic rho-dependent termination sites that will be inserted into plasmid expression vectors. The function of these sites will be evaluated in purified in vitro transcription reactions as well as in vivo.

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University of New Mexico
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