U.S.-Korea Cooperative Research on DNA Sequence Effects on Open Complex Formation by RNA Polymerase

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This proposal requests funds to permit Dr. M. Thomas Record, Department of Chemistry, University of Wisconsin, to pursue with Dr. Jung-Hye Roe, Department of Microbiology, Seoul National University, for a period of 24 months, a program of cooperative research on DNA sequence effects on open complex formation by RNA polymerase. The effect of sequence variations near the transcriptional start site on the rate of formation and dissociation of the open complex will be studied. The three-step mechanism of open-complex formation, proposed by the collaborators in previous research, will be examined in an attempt to identify intermediate steps. In particular, the effects of sequence variations on the rates of association and dissociation under various solution conditions will be studied. In prokaryotes the extent and the timing of gene expression is regulated primarily at the level of transcription initiation. Transcriptional control requires the interaction of specific proteins with specific sites on DNA. In the present research, site-specific mutagenesis of E. coli DNA will be employed to study the effect of the _ ____ introduction of variant promoters on the rates of formation and dissociation of open complexes of RNA polymerase under different conditions of temperature, concentrations of cations and anions, etc. Drs. Record and Roe are highly respected scientists who have successfully collaborated previously in this field of research. This project is relevant to the objectives of the U.S.-Korea Cooperative Science Program which seeks to increase the level of cooperation between U.S. and Korean scientists and engineers through the exchange of scientific information, ideas, skills, and techniques and through collaboration on problems of mutual benefit.