Arginine-deprivation–induced oxidative damage sterilizes <i>Mycobacterium tuberculosis</i>

Abstract

Reactive oxygen species (ROS)-mediated oxidative stress and DNA damage have recently been recognized as contributing to the efficacy of most bactericidal antibiotics, irrespective of their primary macromolecular targets. Inhibitors of targets involved in both combating oxidative stress as well as being required for in vivo survival may exhibit powerful synergistic action. This study demonstrates that the de novo arginine biosynthetic pathway in <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>) is up-regulated in the early response to the oxidative stress-elevating agent isoniazid or vitamin C. Arginine deprivation rapidly sterilizes the <i>Mtb</i> de novo arginine biosynthesis pathway mutants Δ<i>argB</i> and Δ<i>argF</i> without the emergence of suppressor mutants in vitro as well as in vivo. Transcriptomic and flow cytometry studies of arginine-deprived <i>Mtb</i> have indicated accumulation of ROS and extensive DNA damage. Metabolomics studies following arginine deprivation have revealed that these cells experienced depletion of antioxidant thiols and accumulation of the upstream metabolite substrate of ArgB or ArgF enzymes. Δ<i>argB</i> and Δ<i>argF</i> were unable to scavenge host arginine and were quickly cleared from both immunocompetent and immunocompromised mice. In summary, our investigation revealed in vivo essentiality of the de novo arginine biosynthesis pathway for <i>Mtb</i> and a promising drug target space for combating tuberculosis.