title: Starvation, together with the SOS response, mediates high biofilm-specific tolerance to the fluoroquinolone ofloxacin.
abstract: High levels of antibiotic tolerance are a hallmark of bacterial biofilms. In contrast to well-characterized inherited antibiotic resistance, molecular mechanisms leading to reversible and transient antibiotic tolerance displayed by biofilm bacteria are still poorly understood. The physiological heterogeneity of biofilms influences the formation of transient specialized subpopulations that may be more tolerant to antibiotics. In this study, we used random transposon mutagenesis to identify biofilm-specific tolerant mutants normally exhibited by subpopulations located in specialized niches of heterogeneous biofilms. Using Escherichia coli as a model organism, we demonstrated, through identification of amino acid auxotroph mutants, that starved biofilms exhibited significantly greater tolerance towards fluoroquinolone ofloxacin than their planktonic counterparts. We demonstrated that the biofilm-associated tolerance to ofloxacin was fully dependent on a functional SOS response upon starvation to both amino acids and carbon source and partially dependent on the stringent response upon leucine starvation. However, the biofilm-specific ofloxacin increased tolerance did not involve any of the SOS-induced toxin-antitoxin systems previously associated with formation of highly tolerant persisters. We further demonstrated that ofloxacin tolerance was induced as a function of biofilm age, which was dependent on the SOS response. Our results therefore show that the SOS stress response induced in heterogeneous and nutrient-deprived biofilm microenvironments is a molecular mechanism leading to biofilm-specific high tolerance to the fluoroquinolone ofloxacin.
authors.affiliation: Institut Pasteur, Unité de Génétique des Biofilms, Paris, France.
authors.forename: Steve P
authors.initials: SP
authors.lastname: Bernier
authors.forename: David
authors.initials: D
authors.lastname: Lebeaux
authors.forename: Alicia S
authors.initials: AS
authors.lastname: DeFrancesco
authors.forename: Amandine
authors.initials: A
authors.lastname: Valomon
authors.forename: Guillaume
authors.initials: G
authors.lastname: Soubigou
authors.forename: Jean-Yves
authors.initials: JY
authors.lastname: Coppée
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Ghigo
authors.forename: Christophe
authors.initials: C
authors.lastname: Beloin
mesh_terms: D000596:Amino Acids / Q000235:genetics; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D018441:Biofilms / Q000187:drug effects / Q000254:growth & development; D004251:DNA Transposable Elements / Q000235:genetics*; D024881:Drug Resistance, Bacterial / Q000187:drug effects / Q000235:genetics; D004361:Drug Tolerance / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development; D024841:Fluoroquinolones / Q000494:pharmacology; D016296:Mutagenesis; D015242:Ofloxacin / Q000494:pharmacology; D010933:Plankton / Q000187:drug effects / Q000235:genetics; D013014:SOS Response, Genetics; D013217:Starvation
chemical_list: D000596:Amino Acids; D000900:Anti-Bacterial Agents; D004251:DNA Transposable Elements; D024841:Fluoroquinolones; D015242:Ofloxacin