Exposure to sub-inhibitory concentrations of the chemosensitizer 1-(1-naphthylmethyl)-piperazine creates membrane destabilization in multi-drug resistant Klebsiella pneumoniae

João Anes, Sathesh K. Sivasankaran, Dechamma M. Muthappa, Séamus Fanning*, Shabarinath Srikumar

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)
42 Downloads (Pure)

Abstract

Antimicrobial efflux is one of the important mechanisms causing multidrug resistance (MDR) in bacteria. Chemosensitizers like 1-(1-naphthylmethyl)-piperazine (NMP) can inhibit an efflux pump and therefore can overcome MDR. However, secondary effects of NMP other than efflux pump inhibition are rarely investigated. Here, using phenotypic assays, phenotypic microarray and transcriptomic assays we show that NMP creates membrane destabilization in MDR Klebsiella pneumoniae MGH 78578 strain. The NMP mediated membrane destabilization activity was measured using β-lactamase activity, membrane potential alteration studies, and transmission electron microscopy assays. Results from both β-lactamase and membrane potential alteration studies shows that both outer and inner membranes are destabilized in NMP exposed K. pneumoniae MGH 78578 cells. Phenotypic Microarray and RNA-seq were further used to elucidate the metabolic and transcriptional signals underpinning membrane destabilization. Membrane destabilization happens as early as 15 min post-NMP treatment. Our RNA-seq data shows that many genes involved in envelope stress response were differentially regulated in the NMP treated cells. Up-regulation of genes encoding the envelope stress response and repair systems show the distortion in membrane homeostasis during survival in an environment containing sub-inhibitory concentration of NMP. In addition, the lsr operon encoding the production of autoinducer-2 responsible for biofilm production was down-regulated resulting in reduced biofilm formation in NMP treated cells, a phenotype confirmed by crystal violet-based assays. We postulate that the early membrane disruption leads to destabilization of inner membrane potential, impairing ATP production and consequently resulting in efflux pump inhibition.

Original languageEnglish
Article number92
Number of pages14
JournalFrontiers in Microbiology
Volume10
DOIs
Publication statusPublished - 13 Feb 2019

Bibliographical note

Funding Information:
We kindly acknowledge Enterprise Ireland funded Sequencing Alliance for Food Environment (SAFE) project for funding both JA and SS. JA would also like to acknowledge the financial support through the research grant 11/F/051 provided by the Department of Agriculture, Food and the Marine (DAFM), Ireland.

Publisher Copyright:
© 2019 Anes, Sivasankaran, Muthappa, Fanning and Srikumar.

Keywords

  • 1-(1-naphthylmethyl)-piperazine
  • Chemosensitizers
  • Klebsiella pneumoniae
  • Membrane destabilization
  • NMP
  • Secondary effects

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

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