Abstract

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.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI017472-05
Application #
3127233
Study Section
(MG)
Project Start
1980-12-01
Project End
1985-11-30
Budget Start
1984-12-01
Budget End
1985-11-30
Support Year
5
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Public Health Research Institute
Department
Type
DUNS #
City
Newark
State
NJ
Country
United States
Zip Code

  Publications

Hue, K K; Bechhofer, D H (1992) Regulation of the macrolide-lincosamide-streptogramin B resistance gene ermD. J Bacteriol 174:5860-8
Keck, W; van Leeuwen, A M; Huber, M et al. (1990) Cloning and characterization of mepA, the structural gene of the penicillin-insensitive murein endopeptidase from Escherichia coli. Mol Microbiol 4:209-19
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
Villafane, R; Bechhofer, D H; Narayanan, C S et al. (1987) Replication control genes of plasmid pE194. J Bacteriol 169:4822-9
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
Bechhofer, D H; Dubnau, D (1987) Induced mRNA stability in Bacillus subtilis. Proc Natl Acad Sci U S A 84:498-502
Narayanan, C S; Dubnau, D (1987) An in vitro study of the translational attenuation model of ermC regulation. J Biol Chem 262:1756-65
Projan, S J; Monod, M; Narayanan, C S et al. (1987) Replication properties of pIM13, a naturally occurring plasmid found in Bacillus subtilis, and of its close relative pE5, a plasmid native to Staphylococcus aureus. J Bacteriol 169:5131-9
Monod, M; Denoya, C; Dubnau, D (1986) Sequence and properties of pIM13, a macrolide-lincosamide-streptogramin B resistance plasmid from Bacillus subtilis. J Bacteriol 167:138-47

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