Divergent Roles of Escherichia Coli Encoded Lon Protease in Imparting Resistance to Uncouplers of Oxidative Phosphorylation: Roles of marA, rob, soxS and acrB

Verma, Taru; Nandini, Santhi Sanil; Singh, Varsha; Raghavan, Abinaya; Annappa, Harshita; Bhaskarla, Chetana; Dubey, Ashim Kumar; Nandi, Dipankar

doi:10.1007/s00284-024-03632-w

Cited by 1 publication

(1 citation statement)

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“…To study efflux-mediated multidrug resistance, we examined lon -defective mutants of E. coli, which accumulate the MarA protein, subsequently upregulating the AcrAB-TolC efflux pump and rendering them resistant to most antibiotics. − We observed that the PMDR of 4AP-Cn dyes was significantly less in WT bacteria than the Δ lon strain when both were treated with ciprofloxacin and tetracycline, unlike PMB treatment (Figure A). Furthermore, we assessed phenotypic efflux-mediated antibiotic resistance using sodium salicylate (NaSal), a compound that binds to the MarR protein, leading to derepression of MarA expression .…”

Section: Resultsmentioning

confidence: 99%

Quantifying Membrane Alterations with Tailored Fluorescent Dyes: A Rapid Antibiotic Resistance Profiling Methodology

Dubey,

Sardana,

Verma

et al. 2024

ACS Infect. Dis.

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Accurate detection of bacterial antibiotic sensitivity is crucial for theranostics and the containment of antibioticresistant infections. However, the intricate task of detecting and quantifying the antibiotic-induced changes in the bacterial cytoplasmic membrane, and their correlation with other metabolic pathways leading to antibiotic resistance, poses significant challenges. Using a novel class of 4-aminophthalimide (4AP)based fluorescent dyes with precisely tailored alkyl chains, namely 4AP-C9 and 4AP-C13, we quantify stress-mediated alterations in E. coli membranes. Leveraging the unique depth-dependent positioning and environment-sensitive fluorescence properties of these dyes, we detect antibiotic-induced membrane damage through single-cell imaging and monitoring the fluorescence peak maxima difference ratio (PMDR) of the dyes within the bacterial membrane, complemented by other methods. The correlation between the ROS-induced cytoplasmic membrane damage and the PMDR of dyes quantifies sensitivity against bactericidal antibiotics, which correlates to antibiotic-induced lipid peroxidation. Significantly, our findings largely extend to clinical isolates of E. coli and other ESKAPE pathogens like K. pneumoniae and Enterobacter subspecies. Our data reveal that 4AP-Cn probes can potentially act as precise scales to detect antibiotic-induced membrane damage ("thinning") occurring at a subnanometer scale through the quantification of dyes' PMDR, making them promising membrane dyes for rapid detection of bacterial antibiotic resistance, distinguishing sensitive and resistant infections with high specificity in a clinical setup.

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