Target (MexB)- and Efflux-Based Mechanisms Decreasing the Effectiveness of the Efflux Pump Inhibitor D13-9001 in Pseudomonas aeruginosa PAO1: Uncovering a New Role for MexMN-OprM in Efflux of β-Lactams and a Novel Regulatory Circuit (MmnRS) Controlling MexMN Expression

Efflux pumps contribute to antibiotic resistance in Gram-negative pathogens. Correspondingly, efflux pump inhibitors (EPIs) may reverse this resistance. D13-9001 specifically inhibits MexAB-OprM in Mutants with decreased susceptibility to MexAB-OprM inhibition by D13-9001 were identified, and these...

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Published in:Antimicrobial agents and chemotherapy Vol. 63; no. 2
Main Authors: Ranjitkar, Srijan, Jones, Adriana K, Mostafavi, Mina, Zwirko, Zachary, Iartchouk, Oleg, Barnes, S Whitney, Walker, John R, Willis, Thomas W, Lee, Patrick S, Dean, Charles R
Format: Journal Article
Language:English
Published: United States American Society for Microbiology 01-02-2019
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Abstract Efflux pumps contribute to antibiotic resistance in Gram-negative pathogens. Correspondingly, efflux pump inhibitors (EPIs) may reverse this resistance. D13-9001 specifically inhibits MexAB-OprM in Mutants with decreased susceptibility to MexAB-OprM inhibition by D13-9001 were identified, and these fell into two categories: those with alterations in the target MexB (F628L and ΔV177) and those with an alteration in a putative sensor kinase of unknown function, PA1438 (L172P). The alterations in MexB were consistent with reported structural studies of the D13-9001 interaction with MexB. The PA1438 alteration mediated a >150-fold upregulation of MexMN pump gene expression and a >50-fold upregulation of PA1438 and the neighboring response regulator gene, PA1437. We propose that these be renamed and for ex egulator and ex sensor, respectively. MexMN was shown to partner with the outer membrane channel protein OprM and to pump several β-lactams, monobactams, and tazobactam. Upregulated MexMN functionally replaced MexAB-OprM to efflux these compounds but was insusceptible to inhibition by D13-9001. MmnS also mediated a decrease in susceptibility to imipenem and biapenem that was independent of MexMN-OprM. Expression of , encoding the uptake channel for these compounds, was downregulated, suggesting that this channel is also part of the MmnSR regulon. Transcriptome sequencing (RNA-seq) of cells encoding MmnS revealed, among other things, an interrelationship between the regulation of and genes involved in heavy metal resistance.
AbstractList Efflux pumps contribute to antibiotic resistance in Gram-negative pathogens. Correspondingly, efflux pump inhibitors (EPIs) may reverse this resistance. D13-9001 specifically inhibits MexAB-OprM in Mutants with decreased susceptibility to MexAB-OprM inhibition by D13-9001 were identified, and these fell into two categories: those with alterations in the target MexB (F628L and ΔV177) and those with an alteration in a putative sensor kinase of unknown function, PA1438 (L172P). The alterations in MexB were consistent with reported structural studies of the D13-9001 interaction with MexB. The PA1438 alteration mediated a >150-fold upregulation of MexMN pump gene expression and a >50-fold upregulation of PA1438 and the neighboring response regulator gene, PA1437. We propose that these be renamed and for ex egulator and ex sensor, respectively. MexMN was shown to partner with the outer membrane channel protein OprM and to pump several β-lactams, monobactams, and tazobactam. Upregulated MexMN functionally replaced MexAB-OprM to efflux these compounds but was insusceptible to inhibition by D13-9001. MmnS also mediated a decrease in susceptibility to imipenem and biapenem that was independent of MexMN-OprM. Expression of , encoding the uptake channel for these compounds, was downregulated, suggesting that this channel is also part of the MmnSR regulon. Transcriptome sequencing (RNA-seq) of cells encoding MmnS revealed, among other things, an interrelationship between the regulation of and genes involved in heavy metal resistance.
Efflux pumps contribute to antibiotic resistance in Gram-negative pathogens. Correspondingly, efflux pump inhibitors (EPIs) may reverse this resistance. Efflux pumps contribute to antibiotic resistance in Gram-negative pathogens. Correspondingly, efflux pump inhibitors (EPIs) may reverse this resistance. D13-9001 specifically inhibits MexAB-OprM in Pseudomonas aeruginosa . Mutants with decreased susceptibility to MexAB-OprM inhibition by D13-9001 were identified, and these fell into two categories: those with alterations in the target MexB (F628L and ΔV177) and those with an alteration in a putative sensor kinase of unknown function, PA1438 (L172P). The alterations in MexB were consistent with reported structural studies of the D13-9001 interaction with MexB. The PA1438 L172P alteration mediated a >150-fold upregulation of MexMN pump gene expression and a >50-fold upregulation of PA1438 and the neighboring response regulator gene, PA1437. We propose that these be renamed mmnR and mmnS for M ex MN r egulator and M ex MN sensor, respectively. MexMN was shown to partner with the outer membrane channel protein OprM and to pump several β-lactams, monobactams, and tazobactam. Upregulated MexMN functionally replaced MexAB-OprM to efflux these compounds but was insusceptible to inhibition by D13-9001. MmnS L172P also mediated a decrease in susceptibility to imipenem and biapenem that was independent of MexMN-OprM. Expression of oprD , encoding the uptake channel for these compounds, was downregulated, suggesting that this channel is also part of the MmnSR regulon. Transcriptome sequencing (RNA-seq) of cells encoding MmnS L172P revealed, among other things, an interrelationship between the regulation of mexMN and genes involved in heavy metal resistance.
Efflux pumps contribute to antibiotic resistance in Gram-negative pathogens. Correspondingly, efflux pump inhibitors (EPIs) may reverse this resistance. D13-9001 specifically inhibits MexAB-OprM in Pseudomonas aeruginosa. Mutants with decreased susceptibility to MexAB-OprM inhibition by D13-9001 were identified, and these fell into two categories: those with alterations in the target MexB (F628L and ΔV177) and those with an alteration in a putative sensor kinase of unknown function, PA1438 (L172P). The alterations in MexB were consistent with reported structural studies of the D13-9001 interaction with MexB. The PA1438L172P alteration mediated a >150-fold upregulation of MexMN pump gene expression and a >50-fold upregulation of PA1438 and the neighboring response regulator gene, PA1437. We propose that these be renamed mmnR and mmnS for MexMN regulator and MexMN sensor, respectively. MexMN was shown to partner with the outer membrane channel protein OprM and to pump several β-lactams, monobactams, and tazobactam. Upregulated MexMN functionally replaced MexAB-OprM to efflux these compounds but was insusceptible to inhibition by D13-9001. MmnSL172P also mediated a decrease in susceptibility to imipenem and biapenem that was independent of MexMN-OprM. Expression of oprD, encoding the uptake channel for these compounds, was downregulated, suggesting that this channel is also part of the MmnSR regulon. Transcriptome sequencing (RNA-seq) of cells encoding MmnSL172P revealed, among other things, an interrelationship between the regulation of mexMN and genes involved in heavy metal resistance.
Author Jones, Adriana K
Ranjitkar, Srijan
Lee, Patrick S
Walker, John R
Zwirko, Zachary
Willis, Thomas W
Barnes, S Whitney
Mostafavi, Mina
Dean, Charles R
Iartchouk, Oleg
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  givenname: Adriana K
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  givenname: Oleg
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  organization: Infectious Diseases, Novartis Institutes for Biomedical Research, Emeryville, California, USA
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  givenname: Charles R
  orcidid: 0000-0001-9858-9818
  surname: Dean
  fullname: Dean, Charles R
  email: charlesr.dean@novartis.com
  organization: Infectious Diseases, Novartis Institutes for Biomedical Research, Emeryville, California, USA charlesr.dean@novartis.com
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Keywords β-lactams
Pseudomonas aeruginosa
efflux pump inhibitor
drug resistance mechanisms
heavy metals
efflux pumps
Language English
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All Rights Reserved. https://doi.org/10.1128/ASMCopyrightv2
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S.R. and A.K.J. contributed equally to this article.
Citation Ranjitkar S, Jones AK, Mostafavi M, Zwirko Z, Iartchouk O, Barnes SW, Walker JR, Willis TW, Lee PS, Dean CR. 2019. Target (MexB)- and efflux-based mechanisms decreasing the effectiveness of the efflux pump inhibitor D13-9001 in Pseudomonas aeruginosa PAO1: uncovering a new role for MexMN-OprM in efflux of β-lactams and a novel regulatory circuit (MmnRS) controlling MexMN expression. Antimicrob Agents Chemother 63:e01718-18. https://doi.org/10.1128/AAC.01718-18.
Present address: Srijan Ranjitkar, Epizyme, Cambridge, Massachusetts, USA.
ORCID 0000-0001-9858-9818
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PublicationTitle Antimicrobial agents and chemotherapy
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Snippet Efflux pumps contribute to antibiotic resistance in Gram-negative pathogens. Correspondingly, efflux pump inhibitors (EPIs) may reverse this resistance....
Efflux pumps contribute to antibiotic resistance in Gram-negative pathogens. Correspondingly, efflux pump inhibitors (EPIs) may reverse this resistance. Efflux...
SourceID pubmedcentral
proquest
crossref
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pubmed
SourceType Open Access Repository
Aggregation Database
Index Database
SubjectTerms beta-Lactams
beta-Lactams - pharmacology
Imipenem - pharmacology
Mechanisms of Resistance
Microbial Sensitivity Tests
Monobactams - pharmacology
Piperidines
Piperidines - pharmacology
Pseudomonas aeruginosa
Pseudomonas aeruginosa - drug effects
Pseudomonas aeruginosa - genetics
Quaternary Ammonium Compounds
Quaternary Ammonium Compounds - pharmacology
Tazobactam - pharmacology
Thienamycins - pharmacology
Transcriptome - genetics
Title Target (MexB)- and Efflux-Based Mechanisms Decreasing the Effectiveness of the Efflux Pump Inhibitor D13-9001 in Pseudomonas aeruginosa PAO1: Uncovering a New Role for MexMN-OprM in Efflux of β-Lactams and a Novel Regulatory Circuit (MmnRS) Controlling MexMN Expression
URI https://www.ncbi.nlm.nih.gov/pubmed/30420483
https://journals.asm.org/doi/10.1128/AAC.01718-18
https://search.proquest.com/docview/2132734992
https://pubmed.ncbi.nlm.nih.gov/PMC6355610
Volume 63
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