title: Micrococcin P1 - A bactericidal thiopeptide active against Mycobacterium tuberculosis.
abstract: The lack of proper treatment for serious infectious diseases due to the emergence of multidrug resistance reinforces the need for the discovery of novel antibiotics. This is particularly true for tuberculosis (TB) for which 3.7% of new cases and 20% of previously treated cases are estimated to be caused by multi-drug resistant strains. In addition, in the case of TB, which claimed 1.5 million lives in 2014, the treatment of the least complicated, drug sensitive cases is lengthy and disagreeable. Therefore, new drugs with novel targets are urgently needed to control resistant Mycobacterium tuberculosis strains. In this manuscript we report the characterization of the thiopeptide micrococcin P1 as an anti-tubercular agent. Our biochemical experiments show that this antibiotic inhibits the elongation step of protein synthesis in mycobacteria. We have further identified micrococcin resistant mutations in the ribosomal protein L11 (RplK); the mutations were located in the proline loop at the N-terminus. Reintroduction of the mutations into a clean genetic background, confirmed that they conferred resistance, while introduction of the wild type RplK allele into resistant strains re-established sensitivity. We also identified a mutation in the 23S rRNA gene. These data, in good agreement with previous structural studies suggest that also in M. tuberculosis micrococcin P1 functions by binding to the cleft between the 23S rRNA and the L11 protein loop, thus interfering with the binding of elongation factors Tu and G (EF-Tu and EF-G) and inhibiting protein translocation.
authors.affiliation: Department of Molecular Medicine, University of Padova, Padova, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Degiacomi
authors.affiliation: Queen Mary University of London, London E1 2AD, United Kingdom.
authors.forename: Yoann
authors.initials: Y
authors.lastname: Personne
authors.affiliation: Sanofi-Aventis R&D, Drug Disposition, 69367 Lyon, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Mondésert
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, Uppsala, Sweden.
authors.forename: Xueliang
authors.initials: X
authors.lastname: Ge
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, Uppsala, Sweden.
authors.forename: Chandra Sekhar
authors.initials: CS
authors.lastname: Mandava
authors.affiliation: Ecole Polytechnique Fédérale de Lausanne, Global Health Institute, Lausanne, Switzerland.
authors.forename: Ruben C
authors.initials: RC
authors.lastname: Hartkoorn
authors.affiliation: Department of Molecular Medicine, University of Padova, Padova, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Boldrin
authors.affiliation: Queen Mary University of London, London E1 2AD, United Kingdom.
authors.forename: Pavitra
authors.initials: P
authors.lastname: Goel
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, Uppsala, Sweden.
authors.forename: Kristin
authors.initials: K
authors.lastname: Peisker
authors.affiliation: Ecole Polytechnique Fédérale de Lausanne, Global Health Institute, Lausanne, Switzerland.
authors.forename: Andrej
authors.initials: A
authors.lastname: Benjak
authors.affiliation: Sanofi-Aventis R&D, Drug Disposition, 69367 Lyon, France.
authors.forename: Maria Belén
authors.initials: MB
authors.lastname: Barrio
authors.affiliation: Department of Molecular Medicine, University of Padova, Padova, Italy.
authors.forename: Marcello
authors.initials: M
authors.lastname: Ventura
authors.affiliation: Queen Mary University of London, London E1 2AD, United Kingdom.
authors.forename: Amanda C
authors.initials: AC
authors.lastname: Brown
authors.affiliation: Sanofi-Aventis R&D, Drug Disposition, 69367 Lyon, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Leblanc
authors.affiliation: Sanofi-Aventis R&D, Drug Disposition, 69367 Lyon, France.
authors.forename: Armin
authors.initials: A
authors.lastname: Bauer
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, Uppsala, Sweden.
authors.forename: Suparna
authors.initials: S
authors.lastname: Sanyal
authors.affiliation: Ecole Polytechnique Fédérale de Lausanne, Global Health Institute, Lausanne, Switzerland.
authors.forename: Stewart T
authors.initials: ST
authors.lastname: Cole
authors.affiliation: Sanofi-Aventis R&D, Drug Disposition, 69367 Lyon, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Lagrange
authors.affiliation: Queen Mary University of London, London E1 2AD, United Kingdom.
authors.forename: Tanya
authors.initials: T
authors.lastname: Parish
authors.affiliation: Department of Molecular Medicine, University of Padova, Padova, Italy. Electronic address: riccardo.manganelli@unipd.it.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Manganelli
mesh_terms: D000818:Animals; D000904:Antibiotics, Antitubercular / Q000008:administration & dosage / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000096:biosynthesis; D001430:Bacteriocins / Q000008:administration & dosage / Q000494:pharmacology*; D002478:Cells, Cultured; D015169:Colony Count, Microbial; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery; D004353:Drug Evaluation, Preclinical / Q000379:methods; D024881:Drug Resistance, Bacterial / Q000235:genetics; D006801:Humans; D008264:Macrophages / Q000382:microbiology; D008826:Microbial Sensitivity Tests / Q000379:methods; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D010441:Peptide Chain Elongation, Translational / Q000187:drug effects; D010455:Peptides / Q000008:administration & dosage / Q000494:pharmacology*; D012269:Ribosomal Proteins / Q000235:genetics
chemical_list: D000904:Antibiotics, Antitubercular; D001426:Bacterial Proteins; D001430:Bacteriocins; D010455:Peptides; D012269:Ribosomal Proteins; C027624:ribosomal protein L11; C009779:micrococcin
keywords: Drug development; Thiopeptides; Translation; Tuberculosis