000225082 001__ 225082
000225082 005__ 20181203024533.0
000225082 0247_ $$2doi$$a10.1128/Aac.01523-16
000225082 022__ $$a0066-4804
000225082 02470 $$2ISI$$a000389063500002
000225082 037__ $$aARTICLE
000225082 245__ $$aCharacterization of DprE1-Mediated Benzothiazinone Resistance in Mycobacterium tuberculosis
000225082 260__ $$bAmerican Society for Microbiology$$c2016$$aWashington
000225082 269__ $$a2016
000225082 300__ $$a9
000225082 336__ $$aJournal Articles
000225082 520__ $$aBenzothiazinones (BTZs) are a class of compounds found to be extremely potent against both drug-susceptible and drug-resistant Mycobacterium tuberculosis strains. The potency of BTZs is explained by their specificity for their target decaprenylphosphoryl-D-ribose oxidase (DprE1), in particular by covalent binding of the activated form of the compound to the critical cysteine 387 residue of the enzyme. To probe the role of C387, we used promiscuous site-directed mutagenesis to introduce other codons at this position into dprE1 of M. tuberculosis. The resultant viable BTZ-resistant mutants were characterized in vitro, ex vivo, and biochemically to gain insight into the effects of these mutations on DprE1 function and on M. tuberculosis. Five different mutations (C387G, C387A, C387S, C387N, and C387T) conferred various levels of resistance to BTZ and exhibited different phenotypes. The C387G and C387N mutations resulted in a lower growth rate of the mycobacterium on solid medium, which could be attributed to the significant decrease in the catalytic efficiency of the DprE1 enzyme. All five mutations rendered the mycobacterium less cytotoxic to macrophages. Finally, differences in the potencies of covalent and noncovalent DprE1 inhibitors in the presence of C387 mutations were revealed by enzymatic assays. As expected from the mechanism of action, the covalent inhibitor PBTZ169 only partially inhibited the mutant DprE1 enzymes compared to the near-complete inhibition with a noncovalent DprE1 inhibitor, Ty38c. This study emphasizes the importance of the C387 residue for DprE1 activity and for the killing action of covalent inhibitors such as BTZs and other recently identified nitroaromatic inhibitors.
000225082 700__ $$uEcole Polytech Fed Lausanne, Global Hlth Inst, Lausanne, Switzerland$$aFoo, Caroline Shi-Yan
000225082 700__ $$0244938$$g196489$$uEcole Polytech Fed Lausanne, Global Hlth Inst, Lausanne, Switzerland$$aLechartier, Benoit
000225082 700__ $$uEcole Polytech Fed Lausanne, Global Hlth Inst, Lausanne, Switzerland$$aKolly, Gaelle S.
000225082 700__ $$uEcole Polytech Fed Lausanne, Global Hlth Inst, Lausanne, Switzerland$$aBoy-Rottger, Stefanie
000225082 700__ $$0242076$$g196878$$uEcole Polytech Fed Lausanne, Global Hlth Inst, Lausanne, Switzerland$$aNeres, Joao
000225082 700__ $$uEcole Polytech Fed Lausanne, Global Hlth Inst, Lausanne, Switzerland$$aRybniker, Jan
000225082 700__ $$uEcole Polytech Fed Lausanne, Global Hlth Inst, Lausanne, Switzerland$$aLupien, Andreanne
000225082 700__ $$0244366$$g179660$$uEcole Polytech Fed Lausanne, Global Hlth Inst, Lausanne, Switzerland$$aSala, Claudia
000225082 700__ $$uEcole Polytech Fed Lausanne, Global Hlth Inst, Lausanne, Switzerland$$aPiton, Jeremie
000225082 700__ $$aCole, Stewart T.$$uEcole Polytech Fed Lausanne, Global Hlth Inst, Lausanne, Switzerland$$g177247$$0243892
000225082 773__ $$j60$$tAntimicrobial Agents And Chemotherapy$$k11$$q6451-6459
000225082 909C0 $$xU11742$$0252302$$pUPCOL
000225082 909CO $$pSV$$particle$$ooai:infoscience.tind.io:225082
000225082 917Z8 $$x217823
000225082 937__ $$aEPFL-ARTICLE-225082
000225082 973__ $$rREVIEWED$$sPUBLISHED$$aEPFL
000225082 980__ $$aARTICLE