title: Salicylate functions as an efflux pump inducer and promotes the emergence of fluoroquinolone-resistant Campylobacter jejuni mutants.
abstract: Salicylate, a nonsteroidal anti-inflammatory compound, has been shown to increase the resistance of Campylobacter to antimicrobials. However, the molecular mechanism underlying salicylate-induced resistance has not yet been established. In this study, we determined how salicylate increases antibiotic resistance and evaluated its impact on the development of fluoroquinolone-resistant Campylobacter mutants. Transcriptional fusion assays, real-time quantitative reverse transcription-PCR (RT-PCR), and immunoblotting assays consistently demonstrated the induction of the CmeABC multidrug efflux pump by salicylate. Electrophoretic mobility shift assays further showed that salicylate inhibits the binding of CmeR (a transcriptional repressor of the TetR family) to the promoter DNA of cmeABC, suggesting that salicylate inhibits the function of CmeR. The presence of salicylate in the culture medium not only decreased the susceptibility of Campylobacter to ciprofloxacin but also resulted in an approximately 70-fold increase in the observed frequency of emergence of fluoroquinolone-resistant mutants under selection with ciprofloxacin. Together, these results indicate that in Campylobacter, salicylate inhibits the binding of CmeR to the promoter DNA and induces expression of cmeABC, resulting in decreased susceptibility to antibiotics and in increased emergence of fluoroquinolone-resistant mutants under selection pressure.
authors.affiliation: Department of Veterinary Microbiology and Preventive Medicine, 1116 Veterinary Medicine Complex, Iowa State University, Ames, IA 50011, USA.
authors.forename: Zhangqi
authors.initials: Z
authors.lastname: Shen
authors.forename: Xiao-Ying
authors.initials: XY
authors.lastname: Pu
authors.forename: Qijing
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism*; D019677:Artificial Gene Fusion; D001693:Biological Transport, Active / Q000187:drug effects*; D016123:Campylobacter jejuni / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D004269:DNA, Bacterial / Q000378:metabolism; D024881:Drug Resistance, Bacterial; D024202:Electrophoretic Mobility Shift Assay; D024841:Fluoroquinolones / Q000494:pharmacology*; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D015151:Immunoblotting; D011485:Protein Binding; D012097:Repressor Proteins / Q000037:antagonists & inhibitors; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012459:Salicylates / Q000378:metabolism*; D015533:Transcriptional Activation / Q000187:drug effects
chemical_list: D000900:Anti-Bacterial Agents; D000894:Anti-Inflammatory Agents, Non-Steroidal; D004269:DNA, Bacterial; D024841:Fluoroquinolones; D012097:Repressor Proteins; D012459:Salicylates