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Methods to Inhibit Bacterial Pyomelanin Production and Determine the Corresponding Increase in Sensitivity to Oxidative Stress
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NAD+ auxotrophy is bacteriocidal for the tubercle bacilli.

Catherine Vilchèze1, Brian Weinrick, Ka-Wing Wong

  • 1Howard Hughes Medical Institute, Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.

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Mycobacterium bovis possesses a functional salvage pathway for NAD(+) synthesis, unlike previously thought. This allows it to acquire nicotinamide, crucial for growth and distinguishing it from Mycobacterium tuberculosis.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • Mycobacterium tuberculosis and Mycobacterium bovis synthesize NAD(+) via de novo or salvage pathways.
  • A mutation in PncA in M. bovis reportedly impairs its salvage pathway, preventing nicotinic acid secretion and distinguishing it from M. tuberculosis via the niacin test.

Purpose of the Study:

  • To investigate the NAD(+) biosynthesis pathways in Mycobacterium bovis.
  • To determine the role of the salvage pathway in M. bovis growth and nicotinamide acquisition.
  • To assess the potential of targeting common NAD(+) biosynthesis enzymes as drug targets.

Main Methods:

  • Genetic deletion of the de novo biosynthesis pathway (nadABC) in M. bovis and M. tuberculosis.
  • In vivo growth studies of wild-type and mutant strains in mice.
  • Complementation studies by introducing M. tuberculosis pncA into M. bovis DeltanadABC.

Main Results:

  • M. bovis DeltanadABC can be generated, indicating a functional salvage pathway.
  • M. bovis DeltanadABC fails to grow in mice, while M. tuberculosis DeltanadABC grows normally.
  • M. tuberculosis can acquire nicotinamide from the host, utilizing its salvage pathway.
  • Introduction of M. tuberculosis pncA restores growth of M. bovis DeltanadABC in mice, confirming the salvage pathway's role.

Conclusions:

  • NAD(+) starvation is lethal to tubercle bacilli.
  • The functional salvage pathway allows nicotinamide acquisition by M. tuberculosis.
  • Enzymes common to both de novo and salvage NAD(+) pathways are potential drug targets for tuberculosis treatment.