Direct observation of substrate induction of resistance mechanism in Pseudomonas aeruginosa using single live cell imaging

Biochemical and Biophysical Research Communications
Xiao-Hong Nancy XuMichelle E Nowak

Abstract

The resistance mechanism in three strains of Pseudomonas aeruginosa, Delta ABM (devoid of MexAB-OprM), WT, and nalB-1 (overexpression of MexAB-OprM), was investigated using real-time single live cell imaging and fluorescence spectroscopy. Time courses of fluorescence intensity of these three strains in ethidium bromide (EtBr) showed that accumulation kinetics and extrusion machinery were highly dependent upon pump substrate (EtBr) concentration. At high substrate concentration (100 microM), the accumulation kinetic profiles in the cells at earlier incubation times were similar to those observed in low concentration. As EtBr accumulated in the cells reached a critical concentration, the fluorescence intensity of Delta ABM decreased below the fluorescence intensity of EtBr in buffer solution. This result suggested an inductive mechanism in the development of substrate resistance in P. aeruginosa. Substrates appeared to trigger the degradation of EtBr in Delta ABM. Unlike bulk measurements, single live cell imaging overcame the ensemble measurement of bulk analysis and showed that efflux machinery and resistance mechanism in individual cells were not synchronized.

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Citations

Jun 15, 2010·Analytical and Bioanalytical Chemistry·Kerry J LeeXiao-Hong Nancy Xu
Dec 20, 2011·Environmental Science & Technology·Kamila Czechowska, Jan Roelof van der Meer
Mar 24, 2004·Nature Reviews. Microbiology·Mary E Lidstrom, Deirdre R Meldrum
Nov 8, 2002·Journal of Biomedical Optics·Sophia V KyriacouXiao-Hong Nancy Xu
Jul 15, 2009·Biotechnology and Applied Biochemistry·Sheetal RainaTajalli Keshavarz
Jul 3, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Sandra SakalauskaitėRimantas Daugelavičius

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