We will pursue aspects of the molecular biology of pE194, a 3500 base pair plasmid in Bacillus subtilis which confers resistance inducibly to the macrolide-lincosamide-streptogramin B antibiotics. The resistance is conferred via a specific N-methylation of adenine in 23S ribosomal RNA. We will continue studies on the post-transcriptional regulation of synthesis of a protein required for this resistance. Regulation is accompanied by adjustments in the stability of the mRNA for this protein. We will purify and characterize an rRNA methylase specified by pE194. We will pursue studies on gene-overlap in the pE194 genome. These studies include DNA, protein and RNA sequencing and the study of plasmid mutants. Finally, we will continue to characterize DNA replication control in pE194 focussing on defining and studying those regions of the genome which are required for replication and to produce a plasmid-specific trans-acting replication substance. We hope to eventually sequence the entire plasmid genome.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM037137-08
Application #
3292209
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1985-12-01
Project End
1990-11-30
Budget Start
1987-12-01
Budget End
1988-11-30
Support Year
8
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Public Health Research Institute
Department
Type
DUNS #
City
Newark
State
NJ
Country
United States
Zip Code
07103
Dubnau, D (1991) Genetic competence in Bacillus subtilis. Microbiol Rev 55:395-424
Dubnau, D (1991) The regulation of genetic competence in Bacillus subtilis. Mol Microbiol 5:11-8
Su, S L; Dubnau, D (1990) Binding of Bacillus subtilis ermC' methyltransferase to 23S rRNA. Biochemistry 29:6033-42
Breidt, F; Dubnau, D (1990) Identification of cis-acting sequences required for translational autoregulation of the ermC methylase. J Bacteriol 172:3661-8
Dempsey, L A; Dubnau, D A (1989) Localization of the replication origin of plasmid pE194. J Bacteriol 171:2866-9
Dempsey, L A; Dubnau, D A (1989) Identification of plasmid and Bacillus subtilis chromosomal recombination sites used for pE194 integration. J Bacteriol 171:2856-65
Denoya, C; Dubnau, D (1989) Mono- and dimethylating activities and kinetic studies of the ermC 23 S rRNA methyltransferase. J Biol Chem 264:2615-24
Monod, M; Mohan, S; Dubnau, D (1987) Cloning and analysis of ermG, a new macrolide-lincosamide-streptogramin B resistance element from Bacillus sphaericus. J Bacteriol 169:340-50
Denoya, C D; Dubnau, D (1987) Site and substrate specificity of the ermC 23S rRNA methyltransferase. J Bacteriol 169:3857-60
Narayanan, C S; Dubnau, D (1987) Demonstration of erythromycin-dependent stalling of ribosomes on the ermC leader transcript. J Biol Chem 262:1766-71

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