title: N-methylation of a bactericidal compound as a resistance mechanism in Mycobacterium tuberculosis.
abstract: 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.
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medicine, New York, NY 10021;
authors.forename: Thulasi
authors.initials: T
authors.lastname: Warrier
authors.affiliation: Chemical Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY 10065;
authors.forename: Kanishk
authors.initials: K
authors.lastname: Kapilashrami
authors.affiliation: Platform Technology and Science, GlaxoSmithKline, Stevenage SG1 2NY, United Kingdom;
authors.forename: Argyrides
authors.identifier: 0000-0003-3141-9122
authors.initials: A
authors.lastname: Argyrou
authors.affiliation: Department of Computer Science and Engineering, Texas A&M University, College Station, TX 77843-3474;
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Ioerger
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medicine, New York, NY 10021;
authors.forename: David
authors.initials: D
authors.lastname: Little
authors.affiliation: Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA 01655;
authors.forename: Kenan C
authors.initials: KC
authors.lastname: Murphy
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medicine, New York, NY 10021;
authors.forename: Madhumitha
authors.initials: M
authors.lastname: Nandakumar
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medicine, New York, NY 10021;
authors.forename: Suna
authors.initials: S
authors.lastname: Park
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medicine, New York, NY 10021;
authors.forename: Ben
authors.initials: B
authors.lastname: Gold
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medicine, New York, NY 10021;
authors.forename: Jianjie
authors.initials: J
authors.lastname: Mi
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medicine, New York, NY 10021;
authors.forename: Tuo
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Diseases of the Developing World, GlaxoSmithKline (GSK), 28760 Madrid, Spain;
authors.forename: Eugenia
authors.initials: E
authors.lastname: Meiler
authors.affiliation: Platform Technology and Science, GlaxoSmithKline, Stevenage SG1 2NY, United Kingdom;
authors.forename: Mike
authors.initials: M
authors.lastname: Rees
authors.affiliation: Department of Medicine, Weill Cornell Medicine, New York, NY 10021;
authors.forename: Selin
authors.initials: S
authors.lastname: Somersan-Karakaya
authors.affiliation: Diseases of the Developing World, GlaxoSmithKline (GSK), 28760 Madrid, Spain;
authors.forename: Esther
authors.initials: E
authors.lastname: Porras-De Francisco
authors.affiliation: Diseases of the Developing World, GlaxoSmithKline (GSK), 28760 Madrid, Spain;
authors.forename: Maria
authors.initials: M
authors.lastname: Martinez-Hoyos
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medicine, New York, NY 10021;
authors.forename: Kristin
authors.initials: K
authors.lastname: Burns-Huang
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medicine, New York, NY 10021;
authors.forename: Julia
authors.initials: J
authors.lastname: Roberts
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medicine, New York, NY 10021;
authors.forename: Yan
authors.initials: Y
authors.lastname: Ling
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medicine, New York, NY 10021; Department of Medicine, Weill Cornell Medicine, New York, NY 10021;
authors.forename: Kyu Y
authors.initials: KY
authors.lastname: Rhee
authors.affiliation: Diseases of the Developing World, GlaxoSmithKline (GSK), 28760 Madrid, Spain;
authors.forename: Alfonso
authors.initials: A
authors.lastname: Mendoza-Losana
authors.affiliation: Chemical Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY 10065; Department of Pharmacology, Weill Cornell Medicine, New York, NY 10021.
authors.forename: Minkui
authors.initials: M
authors.lastname: Luo
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medicine, New York, NY 10021; cnathan@med.cornell.edu.
authors.forename: Carl F
authors.initials: CF
authors.lastname: Nathan
mesh_terms: D000995:Antitubercular Agents / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001562:Benzimidazoles / Q000737:chemistry / Q000378:metabolism; D024881:Drug Resistance, Bacterial; D015964:Gene Expression Regulation, Bacterial; D008745:Methylation; D008780:Methyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D015394:Molecular Structure; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000378:metabolism*; D000072417:Protein Domains; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000378:metabolism
chemical_list: D000995:Antitubercular Agents; D001426:Bacterial Proteins; D001562:Benzimidazoles; D012097:Repressor Proteins; D012436:S-Adenosylmethionine; C031000:benzimidazole; D008780:Methyltransferases
keywords: antimicrobial resistance; arabinogalactan synthesis; methyltransferase; transcription factor