August 2, 2016
Compounds were dispensed in a black 384-well low-volume microplate (Greiner Bio-One) using a Hewlett Packard HP D300 digital dispenser (Tecan Group Ltd.)
The rising incidence of antimicrobial resistance (AMR) makes it imperative to understand the underlying mechanisms. Mycobacterium tuberculosis (Mtb) is the single leading cause of death from a bacterial pathogen and estimated to be the leading cause of death from AMR. A pyrido-benzimidazole, 14, was reported to have potent bactericidal activity against Mtb. Here, we isolated multiple Mtb clones resistant to 14. Each had mutations in the putative DNA-binding and dimerization domains of rv2887, a gene encoding a transcriptional repressor of the MarR family. The mutations in Rv2887 led to markedly increased expression of rv0560c. We characterized Rv0560c as an S-adenosyl-L-methionine-dependent methyltransferase that N-methylates 14, abolishing its mycobactericidal activity. An Mtb strain lacking rv0560c became resistant to 14 by mutating decaprenylphosphoryl-β-d-ribose 2-oxidase (DprE1), an essential enzyme in arabinogalactan synthesis; 14 proved to be a nanomolar inhibitor of DprE1, and methylation of 14 by Rv0560c abrogated this activity. Thus, 14 joins a growing list of DprE1 inhibitors that are potently mycobactericidal. Bacterial methylation of an antibacterial agent, 14, catalyzed by Rv0560c of Mtb, is a previously unreported mechanism of AMR.
Warrier, T; Kapilashrami, K; Argyrou, A; Ioerger, TR; Little, D; Murphy, KC; Nandakumar, M; Park, S; Gold, B; Mi, J; Zhang, T; Meiler, E; Rees, M; Somersan-Karakaya, S; Porras-De Francisco, E; Martinez-Hoyos, M; Burns-Huang, K; Roberts, J; Ling, Y; Rhee, KY; Mendoza-Losana, A; Luo, M; Nathan, CF;
Journal: Proc. Natl. Acad. Sci. U.S.A. Pages: E4523-E4530