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Mycobacterium smegmatis RNA polymerase: DNA supercoiling, action of rifampicin and mechanism of rifampicin resistance

Levin, ME and Hatfull, GF (1993) Mycobacterium smegmatis RNA polymerase: DNA supercoiling, action of rifampicin and mechanism of rifampicin resistance. Molecular Microbiology, 8 (2). 277 - 285. ISSN 0950-382X

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We have isolated RNA polymerase from Mycobacterium smegmatis and established conditions for specific transcription initiation in vitro. The M. smegmatis enzyme has a strong dependence on supercoiling of the DNA substrate for transcription from mycobacterial promoters. We also show that RNA polymerase is the target for rifampicin, and that this antibiotic specifically inhibits the transition from synthesis of short oligoribonucleotides to full‐length transcripts. RNA polymerase isolated from a rifampicin‐resistant mutant of M. smegmatis is less sensitive to rifampicin in vitro, confirming that one mechanism of rifampicin resistance in mycobacteria is through alteration of RNA polymerase. This in vitro transcription system provides a simple method for the characterization of gene expression in mycobacteria including the pathogens Mycobacterium tuberculosis, Mycobacterium avium and Mycobacterium leprae. It also provides a system for evaluating potential anti‐mycobacterial drugs. Copyright © 1993, Wiley Blackwell. All rights reserved


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Item Type: Article
Status: Published
CreatorsEmailPitt UsernameORCID
Levin, ME
Hatfull, GFgfh@pitt.eduGFH
Date: 1 January 1993
Date Type: Publication
Journal or Publication Title: Molecular Microbiology
Volume: 8
Number: 2
Page Range: 277 - 285
DOI or Unique Handle: 10.1111/j.1365-2958.1993.tb01572.x
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
ISSN: 0950-382X
MeSH Headings: Bacterial Proteins--antagonists & inhibitors; Bacterial Proteins--isolation & purification; Bacterial Proteins--metabolism; DNA, Bacterial--genetics; DNA, Superhelical--genetics; DNA-Directed RNA Polymerases--antagonists & inhibitors; DNA-Directed RNA Polymerases--isolation & purification; DNA-Directed RNA Polymerases--metabolism; Drug Resistance, Microbial; Heat-Shock Proteins--metabolism; Mycobacterium--drug effects; Mycobacterium--enzymology; Mycobacterium--genetics; Promoter Regions, Genetic; Rifampin--pharmacology; Substrate Specificity; Transcription, Genetic
PubMed ID: 8316080
Date Deposited: 12 Mar 2013 19:03
Last Modified: 02 Feb 2019 15:57


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