In E. coli, the synthesis of ribosomes is controlled primarily at the stage of rRNA transcription initiation. rRNA can account for as much as 50% of instantaneous cellular RNA synthesis. In order to understand the biosynthesis of the protein synthetic apparatus, factors involved in the upstream activation and growth rate dependent regulation of rRNA synthesis are being investigated. The mechanism of rRNA promoter-RNA polymerase association is being studied using kinetic and footprinting methods. This will lead to the identification of the individual steps involved in transcription initiation at the rrnB PL promoter and the factors that influence each step. The specific DNA sequences required for upstream activation and growth rate control will be defined by construction and selection of site mutations, followed by assay of operon fusions in vivo. Finally, the importance of position, orientation, and conformation of the upstream activator sequence (UAS) relative to the promoter and the feasibility of using the UAS to enhance transcription from heterologous promoters will also be examined using operon fusions in vivo.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Modified Research Career Development Award (K04)
Project #
5K04AI000935-04
Application #
3070982
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1988-09-01
Project End
1993-08-31
Budget Start
1991-09-01
Budget End
1992-08-31
Support Year
4
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
Schools of Earth Sciences/Natur
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715