1887

Abstract

Aminoglycoside antibiotics are widely used to treat infections of . The MexXY-OprM efflux pump is an important contributor to aminoglycoside tolerance in reference strains and expression of the genes is repressed by the MexZ repressor protein. Direct investigation of the role of this efflux pump in clinical isolates is relatively limited.

The contribution of MexXY-OprM to aminoglycoside resistance is isolate-specific.

To quantify the role of MexXY-OprM and its repressor, MexZ, in clinical isolates of

The genes were deleted from ten clinical isolates of , and the gene from nine isolates. Antimicrobial susceptibility testing was carried out for commonly used antipseudomonal drugs on the engineered mutants and the isogenic wild-type isolates. RT-qPCR was used to measure expression of the gene.

All but one of the mutants were more susceptible to the clinically used aminoglycosides tobramycin, gentamicin and amikacin but the degree to which susceptibility increased varied greatly between isolates. The mutants were also more susceptible to a fluoroquinolone, ciprofloxacin. In three isolates with functional MexZ, deletion of increased expression of and aminoglycoside tolerance. Conversely, deleting from six clinical isolates with sequence variants had little or no effect on expression of or on aminoglycoside susceptibility, consistent with the variants abolishing MexZ function. Genome analysis showed that over 50 % of 619 clinical isolates had sequence variants predicted to reduce the affinity of MexZ for DNA, likely increasing expression and hence efflux of aminoglycosides.

Our findings show that the interplay between MexXY, MexZ and the level of expression plays an important role in aminoglycoside resistance in clinical isolates of but the magnitude of the contribution of this efflux pump to resistance is isolate-specific.

Funding
This study was supported by the:
  • University of Otago
    • Principle Award Recipient: AswinThacharodi
  • Health Research Council of New Zealand (Award 17/372)
    • Principle Award Recipient: IainL Lamont
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/content/journal/jmm/10.1099/jmm.0.001551
2022-06-16
2022-06-28
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