Development of a suicide gene as a novel approach to killing Mycobacterium tuberculosis.
The increase in multidrug-resistant tuberculosis and high mortality among those co-infected with HIV-1 necessitates new therapeutic approaches directed at Mycobacterium tuberculosis. We hypothesized that a dominant-negative mutation in the DNA-dependent RNA polymerase gene would inhibit transcription of all genes by blocking access of the wild-type enzyme to promoters. An evolutionarily invariant lysine was substituted with arginine by site-directed mutagenesis in the rpoB gene. The dominant-negative rpoB gene product inhibited a transposon-derived kanamycin-resistance gene in both M. smegmatis and M. tuberculosis H37Rv, leading to growth inhibition of the mycobacteria on solid media containing kanamycin. The dominant-negative mutant rpoB gene is a potential suicide gene especially for the treatment of multidrug-resistant tuberculosis once a delivery strategy is also developed.
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Department of Medicine, New York University Medical Center, New York City 10016, USA.
Yie
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Tchou-Wong
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KM
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D015139:Blotting, Southern; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D049454:Firefly Luciferin / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics; D005804:Genes, Lethal; D005818:Genetic Engineering; D015316:Genetic Therapy; D006801:Humans; D007613:Kanamycin Resistance / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D009161:Mycobacterium / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000254:growth & development*; D018088:Tuberculosis, Multidrug-Resistant / Q000628:therapy
D001426:Bacterial Proteins; D049454:Firefly Luciferin; D012321:DNA-Directed RNA Polymerases