In vitro and in vivo fitness costs associated with Mycobacterium tuberculosis RpoB mutation H526D

Future Microbiol. 2017 Jul;12(9):753-765. doi: 10.2217/fmb-2017-0022. Epub 2017 Mar 27.

Abstract

Aim: There is controversy regarding the potential fitness costs of rifampicin (RIF) resistance-conferring mutations in the Mycobacterium tuberculosis (Mtb) rpoB gene. We characterized the pathogenicity of an Mtb RpoB H526D mutant.

Materials & methods: A mutant containing the RpoB H526D mutation was isolated from wild-type Mtb grown on RIF-containing plates and complemented for determination of in vitro and in vivo fitness costs.

Results: The RpoB H526D mutant showed reduced survival relative to control strains during progressive hypoxia and delayed growth following resuscitation from nutrient starvation (p < 0.05), which was associated with reduced expression of the resuscitation-promoting factor genes rpfB, rpfC and rpfE. Relative to the isogenic wild-type strain, the mutant showed significantly attenuated growth and long-term survival as well as reduced inflammation in mouse lungs. Conclusion & future perspective: Our data suggest that RpoB H526D mutation confers a fitness cost during growth-limiting conditions in vitro and in mouse lungs.

Keywords: Mycobacterium tuberculosis; Rpf; inflammation; murine model; nutrient starvation; persistence; progressive hypoxia; resuscitation-promoting factor; rpoB; stringent response; virulence.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antitubercular Agents / pharmacology
  • Bacterial Proteins / genetics*
  • DNA-Directed RNA Polymerases / genetics*
  • Disease Models, Animal
  • Drug Resistance, Bacterial / genetics
  • Genetic Complementation Test
  • Genetic Fitness*
  • Humans
  • Inflammation
  • Lung / microbiology
  • Mice
  • Microbial Sensitivity Tests
  • Microbial Viability / drug effects
  • Mutation*
  • Mycobacterium tuberculosis / drug effects
  • Mycobacterium tuberculosis / genetics*
  • Mycobacterium tuberculosis / growth & development
  • Mycobacterium tuberculosis / pathogenicity
  • Oxygen
  • Real-Time Polymerase Chain Reaction
  • Tuberculosis, Multidrug-Resistant / microbiology
  • Virulence

Substances

  • Antitubercular Agents
  • Bacterial Proteins
  • rpoB protein, Mycobacterium tuberculosis
  • DNA-Directed RNA Polymerases
  • Oxygen