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A genetic platform to investigate the functions of bacterial drug efflux pumps

ABSTRACT: Efflux pumps are a significant challenge for the development of new antibacterial agents. Overcoming efflux requires an in-depth understanding of efflux pump functions, substrate specificities, and the development of inhibitors. However, the complexities of drug efflux networks have limited such studies. To address these challenges, we report the generation of Efflux KnockOut-35 (EKO-35), a highly susceptible Escherichia coli strain lacking 35 efflux pumps. We demonstrate the utility of this strain by constructing an efflux platform consisting of strains individually expressing genes encoding efflux pumps forming tripartite complexes with the outer membrane channel TolC. This platform was profiled against a curated diverse compound collection, which enabled us to define physicochemical properties that contribute to transport. We also show the E. coli drug efflux network is conditionally essential for growth, and that the platform can be used to investigate efflux pump inhibitor specificities and also efflux pump interplay. We believe EKO-35 and the efflux platform will have widespread application for the study of drug efflux.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Escherichia Coli

SUBMITTER: Laura Thompson

LAB HEAD: Georgina Cox

PROVIDER: PXD033975 | Pride | 2022-08-31

REPOSITORIES: Pride

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			Action	DRS
	EKO35_LB_Rep1.raw	Raw
	EKO35_LB_Rep2.raw	Raw
	EKO35_LB_Rep3.raw	Raw
	EKO35_LB_Rep4.raw	Raw
	EKO35_M9_Rep1.raw	Raw

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