Alpha-amylase synthesis in Bacillus subtilis 168 is activated at the end of exponential growth, is apparently not inducible, and is repressed by more-readily-metabolized carbon sources, such as glucose. Temporal activation and catabolite repression of amylase synthesis are distinct, genetically-separable events. We have localized the cis-acting regulatory DNA sequences necessary for proper regulation of amylase synthesis to the amyR region at the 5' end of the amylase gene. We propose experiments to define the DNA sequences within amyR which are involved in amylase regulation and to identify trans-acting factors which interact with these regulatory sites. Specifically, we propose a series of experiments designed to dissect the functional domains of the amyR region using targeted mutagenesis. In addition, we propose to identify and characterize trans-acting regulatory elements through a program combining mutagenesis with studies of physical interactions between trans-regulatory factors and the functional sites within amyR. Elucidation of the mechanism of amylase regulation resulting from these experiments will provide insights into the regulatory mechanisms governing the expression of other postexponentially-expressed genes in B. subtilis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM034324-06
Application #
3285107
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1984-12-01
Project End
1992-12-31
Budget Start
1990-01-01
Budget End
1990-12-31
Support Year
6
Fiscal Year
1990
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
Voskuil, Martin I; Chambliss, Glenn H (2002) The TRTGn motif stabilizes the transcription initiation open complex. J Mol Biol 322:521-32
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