The role of RNA polymerase in the regulation of gene expression during growth and sporulation of Bacillus subtilis will be investigated. Since the sigma-43 form of RNA polymerase plays a major role during growth, early stationary phase, and catabolite repression, the following aspects of this enzyme will be investigated in detail: (1) the genetic organization and regulation of the recently cloned and sequenced sigma-43 operon will be analyzed by determining its promoter sites and RNA polymerase specificities, by testing various environmental and physiological conditions that alter transcription from these promoters, by determining the function of the first gene of the operon, and by modifying the promoter regions by site directed mutagenesis; (2) the role of sigma-43 enzyme in catabolite resistant sporulation and catabolite repression will be investigated by studying the relationship between the crsA mutation in the rpoD (sigma-43) gene and the spoO and non-sporulation related suppressor genes for crsA. The studies will involve the isolation of additional suppressor genes for crsA including those which are not related to sporulation. Glucose sensitive promoters will be isolated by a newly developed promoter expression probe plasmid and the effect of the crsA mutation will be tested on the promoters by in vivo and in vitro transcription studies. Those spoO genes which suppress crsA and which can be suppressed by crsA will be cloned and characterized. The effect of spoO gene products will be examined on transcription carried out by the wild type sigma-43 enzyme and the crsA mutated sigma-43 enzyme.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM019673-19
Application #
3269730
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1976-09-01
Project End
1992-08-31
Budget Start
1990-09-01
Budget End
1992-08-31
Support Year
19
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of California Davis
Department
Type
Schools of Earth Sciences/Natur
DUNS #
094878337
City
Davis
State
CA
Country
United States
Zip Code
95618
Chang, B Y; Doi, R H (1993) Conformational properties of Bacillus subtilis RNA polymerase sigma A factor during transcription initiation. Biochem J 294 ( Pt 1):43-7
Chang, B Y; Doi, R H (1993) Effects of amino acid substitutions in the promoter -10 binding region of the sigma A factor on growth of Bacillus subtilis. J Bacteriol 175:2470-4
McCready, P M; Doi, R H (1992) Bacillus subtilis SenS exerts its activity through a site in the 5' flanking region of the aprE promoter. J Gen Microbiol 138:2069-74
Chang, B Y; Shyu, Y T; Doi, R H (1992) The interaction between Bacillus subtilis sigma-A (sigma A) factor and RNA polymerase with promoters. Biochimie 74:601-12
Qi, F X; He, X S; Doi, R H (1991) Localization of a new promoter, P5, in the sigA operon of Bacillus subtilis and its regulation in some spo mutant strains. J Bacteriol 173:7050-4
Foong, F; Hamamoto, T; Shoseyov, O et al. (1991) Nucleotide sequence and characteristics of endoglucanase gene engB from Clostridium cellulovorans. J Gen Microbiol 137:1729-36
He, X S; Shyu, Y T; Nathoo, S et al. (1991) Construction and use of a Bacillus subtilis mutant deficient in multiple protease genes for the expression of eukaryotic genes. Ann N Y Acad Sci 646:69-77
Venezia, N D; Krakow, J S (1990) Effects of anti-alpha monoclonal antibodies on initiation and elongation by the Escherichia coli RNA polymerase. J Biol Chem 265:8122-6
Zuberi, A R; Doi, R H (1990) A mutation in P23, the first gene in the RNA polymerase sigma A (sigma 43) operon, affects sporulation in Bacillus subtilis. J Bacteriol 172:2175-7
Chang, B Y; Doi, R H (1990) Overproduction, purification, and characterization of Bacillus subtilis RNA polymerase sigma A factor. J Bacteriol 172:3257-63

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