Universität zu Köln

Foo, Caroline Shi-Yan ORCID: 0000-0002-6380-4917, Lechartier, Benoit, Kolly, Gaelle S., Boy-Rottger, Stefanie, Neres, Joao ORCID: 0000-0003-4488-2423, Rybniker, Jan, Lupien, Andreanne, Sala, Claudia ORCID: 0000-0003-4031-4648, Piton, Jeremie and Cole, Stewart T. (2016). Characterization of DprE1-Mediated Benzothiazinone Resistance in Mycobacterium tuberculosis. Antimicrob. Agents Chemother., 60 (11). S. 6451 - 6460. WASHINGTON: AMER SOC MICROBIOLOGY. ISSN 1098-6596

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Abstract

Benzothiazinones (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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Foo, Caroline Shi-Yan UNSPECIFIED orcid.org/0000-0002-6380-4917 UNSPECIFIED Lechartier, Benoit UNSPECIFIED UNSPECIFIED UNSPECIFIED Kolly, Gaelle S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Boy-Rottger, Stefanie UNSPECIFIED UNSPECIFIED UNSPECIFIED Neres, Joao UNSPECIFIED orcid.org/0000-0003-4488-2423 UNSPECIFIED Rybniker, Jan UNSPECIFIED UNSPECIFIED UNSPECIFIED Lupien, Andreanne UNSPECIFIED UNSPECIFIED UNSPECIFIED Sala, Claudia UNSPECIFIED orcid.org/0000-0003-4031-4648 UNSPECIFIED Piton, Jeremie UNSPECIFIED UNSPECIFIED UNSPECIFIED Cole, Stewart T. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-257575
DOI:	10.1128/AAC.01523-16
Journal or Publication Title:	Antimicrob. Agents Chemother.
Volume:	60
Number:	11
Page Range:	S. 6451 - 6460
Date:	2016
Publisher:	AMER SOC MICROBIOLOGY
Place of Publication:	WASHINGTON
ISSN:	1098-6596
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language INHIBITORS; DPRE1; COMBINATION; ARABINAN Multiple languages Microbiology; Pharmacology & Pharmacy Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/25757