Anti-Bacterial Agents (therapeutic use); Benzamides (chemistry, therapeutic use); Blotting, Western; Catalytic Domain; Chromatography, Liquid; Dinitrobenzenes (chemistry, therapeutic use); Mycobacterium Infections, Nontuberculous (drug therapy); Mycobacterium smegmatis (drug effects); Nitroreductases (genetics, metabolism); RNA, Messenger (genetics); Racemases and Epimerases (antagonists & inhibitors, genetics, metabolism); Real-Time Polymerase Chain Reaction; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Thiazines (chemistry, therapeutic use); Tuberculosis, Multidrug-Resistant (drug therapy, microbiology)
Tuberculosis is still a leading cause of death worldwide. The selection and spread of Mycobacterium tuberculosis multidrug-resistant (MDR-TB) and extensively drug-resistant strains (XDR-TB) is a severe public health problem. Recently, two different classes of chemical series, the benzothiazinones (BTZ) and the dinitrobenzamide (DNB) derivatives have been found to be highly active against M. tuberculosis, including XDR-TB strains. The target of BTZs is DprE1 protein which works in concert with DprE2 to form the heteromeric decaprenylphosphoryl-β-D-ribose 2'-epimerase, involved in Decaprenyl-Phospho-Arabinose (DPA) biosynthesis. Interestingly, it has been shown that the DNBs block the same pathway thus suggesting that both drugs could share the same target. Moreover, in Mycobacterium smegmatis the overexpression of the NfnB nitroreductase led to the inactivation of the BTZs by reduction of a critical nitro-group to an amino-group. In this work several spontaneous M. smegmatis mutants resistant to DNBs were isolated. Sixteen mutants, showing high levels of DNB resistance, exhibited a mutation in the Cys394 of DprE1. Using fluorescence titration and mass spectrometry it has been possible to monitor the binding between DprE1 and DNBs, achieving direct evidence that MSMEG_6382 is the cellular target of DNBs in mycobacteria. Additionally, M. smegmatis mutants having low levels of resistance to DNBs harbor various mutations in MSMEG_6503 gene encoding the transcriptional repressor of the nitroreductase NfnB. By LC/MS analysis it has been demonstrated that NfnB is responsible for DNB inactivation. Taken together, our data demonstrate that both DNB and BTZ drugs share common resistance mechanisms in M. smegmatis.
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Published in PloS ONE, Volume 6, Issue 11, 2011, pages e26675-.
©2011 de Jesus Lopes Ribeiro et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
de Jesus Lopes Ribeiro, A., Degiacomi, G., Ewann, F., Buroni, S., Incandela, M., & Chiarelli, L. et al. (2011). Analogous Mechanisms of Resistance to Benzothiazinones and Dinitrobenzamides in Mycobacterium smegmatis. Plos ONE, 6(11), e26675. https://doi.org/10.1371/journal.pone.0026675