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Suppression of the recA SOS response gene and reactive oxygen species (ROS) overproduction have been shown, separately, to enhance fluoroquinolone activity and lethality. Their putative synergistic impact as a strategy to potentiate the efficacy of bactericidal antimicrobial agents such as fluoroquinolones is unknown. We generated Escherichia coli mutants that exhibited a suppressed Δ recA gene in combination with inactivated ROS detoxification system genes (Δ sodA , Δ sodB , Δ katG , Δ katE , and Δ ahpC ) or inactivated oxidative stress regulator genes (Δ oxyR and Δ rpoS ) to evaluate the interplay of both DNA repair and detoxification systems in drug response. Synergistic sensitization effects, ranging from 7.5- to 30-fold relative to the wild type, were observed with ciprofloxacin in double knockouts of recA and inactivated detoxification system genes. Compared to recA knockout, inactivation of an additional detoxification system gene reduced MIC values up to 8-fold. In growth curves, no growth was evident in mutants doubly deficient for recA gene and oxidative detoxification systems at subinhibitory concentrations of ciprofloxacin, in contrast to the recA -deficient strain. There was a marked reduction of viable bacteria in a short period of time when the recA gene and other detoxification system genes ( katG , sodA , or ahpC ) were inactivated (using absolute ciprofloxacin concentrations). At 4 h, a bactericidal effect of ciprofloxacin was observed for Δ katG Δ recA and Δ ahpC Δ recA double mutants compared to the single Δ recA mutant (Δ3.4 log 10 CFU/ml). Synergistic quinolone sensitization, by targeting the recA gene and oxidative detoxification stress systems, reinforces the role of both DNA repair systems and ROS in antibiotic-induced bacterial cell death, opening up a new pathway for antimicrobial sensitization.

Synergistic Quinolone Sensitization by Targeting the recA SOS Response Gene and Oxidative Stress