Francisella tularensis: FupA mutation contributes to fluoroquinolone resistance by increasing vesicle secretion and biofilm formation

Francisella tularensis is the causative agent in tularemia for which the high prevalence of treatment failure and relapse is a major concern. Directed-evolution experiments revealed that acquisition of fluoroquinolone (FQ) resistance was linked to factors in addition to mutations in DNA gyrase. Here...

Full description

Saved in:
Bibliographic Details
Main Authors: Claire Siebert, Helena Lindgren, Sabrina Ferré, Corinne Villers, Sandrine Boisset, Julien Perard, Anders Sjöstedt, Max Maurin, Céline Brochier-Armanet, Yohann Couté, Patricia Renesto
Format: Article
Language:English
Published: Taylor & Francis Group 2019-01-01
Series:Emerging Microbes and Infections
Subjects:
Online Access:https://www.tandfonline.com/doi/10.1080/22221751.2019.1615848
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850216671129632768
author Claire Siebert
Helena Lindgren
Sabrina Ferré
Corinne Villers
Sandrine Boisset
Julien Perard
Anders Sjöstedt
Max Maurin
Céline Brochier-Armanet
Yohann Couté
Patricia Renesto
author_facet Claire Siebert
Helena Lindgren
Sabrina Ferré
Corinne Villers
Sandrine Boisset
Julien Perard
Anders Sjöstedt
Max Maurin
Céline Brochier-Armanet
Yohann Couté
Patricia Renesto
author_sort Claire Siebert
collection DOAJ
description Francisella tularensis is the causative agent in tularemia for which the high prevalence of treatment failure and relapse is a major concern. Directed-evolution experiments revealed that acquisition of fluoroquinolone (FQ) resistance was linked to factors in addition to mutations in DNA gyrase. Here, using F. tularensis live vaccine strain (LVS) as a model, we demonstrated that FupA/B (Fer-Utilization Protein) expression is linked to FQ susceptibility, and that the virulent strain F. tularensis subsp. tularensis SCHU S4 deleted for the homologous FupA protein exhibited even higher FQ resistance. In addition to an increased FQ minimal inhibitory concentration, LVSΔfupA/B displayed tolerance toward bactericidal compounds including ciprofloxacin and gentamicin. Interestingly, the FupA/B deletion was found to promote increased secretion of outer membrane vesicles (OMVs). Mass spectrometry-based quantitative proteomic characterization of vesicles from LVS and LVS∆fupA/B identified 801 proteins, including a subset of 23 proteins exhibiting differential abundance between both strains which may therefore contribute to the reduced antibiotic susceptibility of the FupA/B-deleted strain. We also demonstrated that OMVs are key structural elements of LVSΔfupA/B biofilms providing protection against FQ. These results provide a new basis for understanding and tackling antibiotic resistance and/or persistence of Francisella and other pathogenic members of the Thiotrichales class.
format Article
id doaj-art-0428d34834be4f76b1cbc00e4e120bd3
institution OA Journals
issn 2222-1751
language English
publishDate 2019-01-01
publisher Taylor & Francis Group
record_format Article
series Emerging Microbes and Infections
spelling doaj-art-0428d34834be4f76b1cbc00e4e120bd32025-08-20T02:08:14ZengTaylor & Francis GroupEmerging Microbes and Infections2222-17512019-01-018180882210.1080/22221751.2019.1615848Francisella tularensis: FupA mutation contributes to fluoroquinolone resistance by increasing vesicle secretion and biofilm formationClaire Siebert0Helena Lindgren1Sabrina Ferré2Corinne Villers3Sandrine Boisset4Julien Perard5Anders Sjöstedt6Max Maurin7Céline Brochier-Armanet8Yohann Couté9Patricia Renesto10TIMC-IMAG UMR 5525 - UGA CNRS, Grenoble, FranceLaboratory for Molecular Infection Medicine Sweden and Department of Clinical Microbiology, Umeå University, Umeå, SwedenUniversité Grenoble Alpes, CEA, Inserm, IRIG-BGE, Grenoble, FranceTIMC-IMAG UMR 5525 - UGA CNRS, Grenoble, FranceTIMC-IMAG UMR 5525 - UGA CNRS, Grenoble, FranceUniversité Grenoble Alpes, CNRS, CEA, BIG-LCBM, Grenoble, FranceLaboratory for Molecular Infection Medicine Sweden and Department of Clinical Microbiology, Umeå University, Umeå, SwedenTIMC-IMAG UMR 5525 - UGA CNRS, Grenoble, FranceLaboratoire de Biométrie et Biologie Évolutive, Université Claude Bernard Lyon 1, CNRS, UMR5558, Villeurbanne, FranceUniversité Grenoble Alpes, CEA, Inserm, IRIG-BGE, Grenoble, FranceTIMC-IMAG UMR 5525 - UGA CNRS, Grenoble, FranceFrancisella tularensis is the causative agent in tularemia for which the high prevalence of treatment failure and relapse is a major concern. Directed-evolution experiments revealed that acquisition of fluoroquinolone (FQ) resistance was linked to factors in addition to mutations in DNA gyrase. Here, using F. tularensis live vaccine strain (LVS) as a model, we demonstrated that FupA/B (Fer-Utilization Protein) expression is linked to FQ susceptibility, and that the virulent strain F. tularensis subsp. tularensis SCHU S4 deleted for the homologous FupA protein exhibited even higher FQ resistance. In addition to an increased FQ minimal inhibitory concentration, LVSΔfupA/B displayed tolerance toward bactericidal compounds including ciprofloxacin and gentamicin. Interestingly, the FupA/B deletion was found to promote increased secretion of outer membrane vesicles (OMVs). Mass spectrometry-based quantitative proteomic characterization of vesicles from LVS and LVS∆fupA/B identified 801 proteins, including a subset of 23 proteins exhibiting differential abundance between both strains which may therefore contribute to the reduced antibiotic susceptibility of the FupA/B-deleted strain. We also demonstrated that OMVs are key structural elements of LVSΔfupA/B biofilms providing protection against FQ. These results provide a new basis for understanding and tackling antibiotic resistance and/or persistence of Francisella and other pathogenic members of the Thiotrichales class.https://www.tandfonline.com/doi/10.1080/22221751.2019.1615848FrancisellaOMVsbiofilmsantibioticsfluoroquinolones
spellingShingle Claire Siebert
Helena Lindgren
Sabrina Ferré
Corinne Villers
Sandrine Boisset
Julien Perard
Anders Sjöstedt
Max Maurin
Céline Brochier-Armanet
Yohann Couté
Patricia Renesto
Francisella tularensis: FupA mutation contributes to fluoroquinolone resistance by increasing vesicle secretion and biofilm formation
Emerging Microbes and Infections
Francisella
OMVs
biofilms
antibiotics
fluoroquinolones
title Francisella tularensis: FupA mutation contributes to fluoroquinolone resistance by increasing vesicle secretion and biofilm formation
title_full Francisella tularensis: FupA mutation contributes to fluoroquinolone resistance by increasing vesicle secretion and biofilm formation
title_fullStr Francisella tularensis: FupA mutation contributes to fluoroquinolone resistance by increasing vesicle secretion and biofilm formation
title_full_unstemmed Francisella tularensis: FupA mutation contributes to fluoroquinolone resistance by increasing vesicle secretion and biofilm formation
title_short Francisella tularensis: FupA mutation contributes to fluoroquinolone resistance by increasing vesicle secretion and biofilm formation
title_sort francisella tularensis fupa mutation contributes to fluoroquinolone resistance by increasing vesicle secretion and biofilm formation
topic Francisella
OMVs
biofilms
antibiotics
fluoroquinolones
url https://www.tandfonline.com/doi/10.1080/22221751.2019.1615848
work_keys_str_mv AT clairesiebert francisellatularensisfupamutationcontributestofluoroquinoloneresistancebyincreasingvesiclesecretionandbiofilmformation
AT helenalindgren francisellatularensisfupamutationcontributestofluoroquinoloneresistancebyincreasingvesiclesecretionandbiofilmformation
AT sabrinaferre francisellatularensisfupamutationcontributestofluoroquinoloneresistancebyincreasingvesiclesecretionandbiofilmformation
AT corinnevillers francisellatularensisfupamutationcontributestofluoroquinoloneresistancebyincreasingvesiclesecretionandbiofilmformation
AT sandrineboisset francisellatularensisfupamutationcontributestofluoroquinoloneresistancebyincreasingvesiclesecretionandbiofilmformation
AT julienperard francisellatularensisfupamutationcontributestofluoroquinoloneresistancebyincreasingvesiclesecretionandbiofilmformation
AT anderssjostedt francisellatularensisfupamutationcontributestofluoroquinoloneresistancebyincreasingvesiclesecretionandbiofilmformation
AT maxmaurin francisellatularensisfupamutationcontributestofluoroquinoloneresistancebyincreasingvesiclesecretionandbiofilmformation
AT celinebrochierarmanet francisellatularensisfupamutationcontributestofluoroquinoloneresistancebyincreasingvesiclesecretionandbiofilmformation
AT yohanncoute francisellatularensisfupamutationcontributestofluoroquinoloneresistancebyincreasingvesiclesecretionandbiofilmformation
AT patriciarenesto francisellatularensisfupamutationcontributestofluoroquinoloneresistancebyincreasingvesiclesecretionandbiofilmformation