Description of compensatory gyrA mutations restoring fluoroquinolone susceptibility in Mycobacterium tuberculosis
| Autor: | Stéphanie Petrella, Alexandra Aubry, Nicolas Veziris, Hélène Ferrand, Claudine Mayer, Aurélie Bouige, Alix Pantel, Stéphanie Matrat, Wladimir Sougakoff |
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| Přispěvatelé: | Université Pierre et Marie Curie - Paris 6 (UPMC), Microbiologie structurale - Structural Microbiology (Microb. Struc. (UMR_3528 / U-Pasteur_5)), Institut Pasteur [Paris] (IP)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Centre d'Immunologie et de Maladies Infectieuses (CIMI), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Parts of this paper were presented at the ‘International Conference on Antimicrobial Agents and Chemotherapy’, USA, 2011 (C1-626), and at the EMBO Conference ‘Tuberculosis 2012’, France, 2012 (TUBERCULOSIS2012/000269)., Institut Pasteur [Paris]-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Centre National de Référence des Mycobactéries et de la Résistance aux Antituberculeux [CHU Pitié-Salpêtrière], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Centre National de Référence des Mycobactéries et de la Résistance des Mycobactéries aux Antituberculeux, Bactériologie-Hygiène [CHU Pitié-Salpêtrière] ], CHU Pitié-Salpêtrière [APHP] |
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
MESH: Mycobacterium tuberculosis DNA Mutational Analysis Mutant Antitubercular Agents medicine.disease_cause DNA gyrase MESH: Recombinant Proteins Suppression Genetic [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases Pharmacology (medical) MESH: DNA Mutational Analysis MESH: Suppression Genetic MESH: Inhibitory Concentration 50 Mutation MESH: Microbial Sensitivity Tests biology Chemistry MESH: Escherichia coli Recombinant Proteins 3. Good health MESH: Mutagenesis Site-Directed Infectious Diseases [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology DNA Gyrase DNA supercoil Fluoroquinolones medicine.drug MESH: DNA Gyrase Microbiology (medical) 030106 microbiology Mutation Missense Microbial Sensitivity Tests Microbiology Mycobacterium tuberculosis Inhibitory Concentration 50 03 medical and health sciences Drug Resistance Bacterial MESH: Drug Resistance Bacterial Escherichia coli medicine Humans Pharmacology MESH: Mutation Missense MESH: Humans Mutagenesis MESH: Fluoroquinolones biology.organism_classification MESH: Antitubercular Agents Mutagenesis Site-Directed [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology Ofloxacin |
| Zdroj: | Journal of Antimicrobial Chemotherapy Journal of Antimicrobial Chemotherapy, 2016, 71 (9), pp.2428-2431. ⟨10.1093/jac/dkw169⟩ Journal of Antimicrobial Chemotherapy, Oxford University Press (OUP), 2016, 71 (9), pp.2428-2431. ⟨10.1093/jac/dkw169⟩ |
| ISSN: | 0305-7453 1460-2091 |
| DOI: | 10.1093/jac/dkw169⟩ |
| Popis: | International audience; OBJECTIVES:Resistance to fluoroquinolones (FQs) in Mycobacterium tuberculosis (Mtb) is mainly due to mutations in DNA gyrase (GyrA2B2), with the most common substitutions located at positions 90 and 94 in GyrA. Two clinical MDR Mtb (MDR-TB) strains harbouring an A90E or D94N substitution in GyrA were found to be surprisingly susceptible to FQs (ofloxacin MIC ≤2 mg/L). We studied the impact of the additional GyrA substitutions found in these strains (T80A and T80A + A90G, respectively) on FQ susceptibility.METHODS:Mutants of interest were generated by site-specific mutagenesis of GyrA alleles. WT and mutant TB DNA gyrase subunits were overexpressed in Escherichia coli and purified, and the in vitro susceptibility to FQs of their DNA supercoiling reaction was studied.RESULTS:IC50s of mutant gyrase complexes bearing GyrA D94N and A90E were 3- to 36-fold higher than WT IC50s, whereas IC50s of gyrase bearing T80A + A90G + D94N and T80A + A90E were close to the WT IC50s.CONCLUSIONS:We demonstrated that substitutions T80A and A90G restore FQ susceptibility when associated with a substitution implicated in high-level FQ resistance. Line probe assay misclassification of MDR-TB strains as pre-XDR or XDR can be corrected by sequence analysis of gyrA. |
| Databáze: | OpenAIRE |
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