A shared mechanism of multidrug resistance in laboratory-evolved uropathogenic .

The emergence of multidrug-resistant bacteria poses a significant threat to human health, necessitating a comprehensive understanding of their underlying mechanisms. Uropathogenic (UPEC), the primary causative agent of urinary tract infections, is frequently associated with multidrug resistance and recurrent infections. To elucidate the mechanism of resistance of UPEC to beta-lactam antibiotics, we generated ampicillin-resistant UPEC strains through continuous exposure to low and high levels of ampicillin in the laboratory, referred to as Low Amp and High Amp, respectively. Whole-genome sequencing revealed that both Low and High Amp strains contained mutations in the , , and genes. The High Amp strain exhibited a single additional mutation in the gene. Using protein modeling and qRT-PCR analyses, we validated the contributions of each mutation in the identified genes to antibiotic resistance in the Amp strains, including a decrease in membrane permeability, increased expression of multidrug efflux pump, and inhibition of cell lysis. Furthermore, the Amp strain does not decrease the bacterial burden in the mouse bladder even after continuous antibiotic treatment , implicating the increasing difficulty in treating host infections caused by the Amp strain. Interestingly, ampicillin-induced mutations also result in multidrug resistance in UPEC, suggesting a common mechanism by which bacteria acquire cross-resistance to other classes of antibiotics.