Inactivation of [Fe-S] metalloproteins mediates nitric oxide-dependent killing of Burkholderia mallei.
BACKGROUND
Much remains to be known about the mechanisms by which O(2)-dependent host defenses mediate broad antimicrobial activity.


RESULTS
We show herein that reactive nitrogen species (RNS) generated by inducible nitric oxide (NO) synthase (iNOS) account for the anti-Burkholderia mallei activity of IFNgamma-primed macrophages. Inducible NOS-mediated intracellular killing may represent direct bactericidal activity, because B. mallei showed an exquisite sensitivity to NO generated chemically. Exposure of B. mallei to sublethal concentrations of NO upregulated transcription of [Fe-S] cluster repair genes, while damaging the enzymatic activity of the [Fe-S] protein aconitase. To test whether [Fe-S] clusters are critical targets for RNS-dependent killing of B. mallei, a mutation was constructed in the NO-induced, [Fe-S] cluster repair regulator iscR. Not only was the iscR mutant hypersusceptible to iNOS-mediated killing, but its aconitase pool was readily oxidized by NO donors as compared to wild-type controls. Although killed by authentic H(2)O(2), which also oxidizes [Fe-S] clusters, B. mallei appear to be resilient to NADPH oxidase-mediated cytotoxicity. The poor respiratory burst elicited by this bacterium likely explains why the NADPH oxidase is nonessential to the killing of B. mallei while it is still confined within phagosomes.


CONCLUSIONS
Collectively, these findings have revealed a disparate role for NADPH oxidase and iNOS in the innate macrophage response against the strict aerobe B. mallei. To the best of our knowledge, this is the first instance in which disruption of [Fe-S] clusters is demonstrated as cause of the bactericidal activity of NO congeners.
Jones-Carson
Jessica
J
Department of Microbiology, University of Colorado Health Sciences Center, Aurora, Colorado, United States of America.
Laughlin
James
J
Hamad
Mohammed A
MA
Stewart
Amanda L
AL
Voskuil
Martin I
MI
Vázquez-Torres
Andrés
A
D000154:Aconitate Hydratase / Q000378:metabolism; D000818:Animals; D042726:Burkholderia mallei / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007506:Iron-Sulfur Proteins / Q000737:chemistry*; D008264:Macrophages / Q000378:metabolism / Q000382:microbiology; D051379:Mice; D008810:Mice, Inbred C57BL; D008954:Models, Biological; D008957:Models, Genetic; D019255:NADPH Oxidases / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism; D010588:Phagosomes / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism
D007506:Iron-Sulfur Proteins; D013481:Superoxides; D009569:Nitric Oxide; D006861:Hydrogen Peroxide; D019255:NADPH Oxidases; D000154:Aconitate Hydratase; D010100:Oxygen