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 |
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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 |
Author_xml | – sequence: 1 givenname: Srijan surname: Ranjitkar fullname: Ranjitkar, Srijan organization: Infectious Diseases, Novartis Institutes for Biomedical Research, Emeryville, California, USA – sequence: 2 givenname: Adriana K surname: Jones fullname: Jones, Adriana K organization: Infectious Diseases, Novartis Institutes for Biomedical Research, Emeryville, California, USA – sequence: 3 givenname: Mina surname: Mostafavi fullname: Mostafavi, Mina organization: Infectious Diseases, Novartis Institutes for Biomedical Research, Emeryville, California, USA – sequence: 4 givenname: Zachary surname: Zwirko fullname: Zwirko, Zachary organization: Biotherapeutic and Analytical Technologies, Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, USA – sequence: 5 givenname: Oleg surname: Iartchouk fullname: Iartchouk, Oleg organization: Biotherapeutic and Analytical Technologies, Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, USA – sequence: 6 givenname: S Whitney surname: Barnes fullname: Barnes, S Whitney organization: Genomics Institute of Novartis Research Foundation (GNF), San Diego, California, USA – sequence: 7 givenname: John R surname: Walker fullname: Walker, John R organization: Genomics Institute of Novartis Research Foundation (GNF), San Diego, California, USA – sequence: 8 givenname: Thomas W surname: Willis fullname: Willis, Thomas W organization: Infectious Diseases, Novartis Institutes for Biomedical Research, Emeryville, California, USA – sequence: 9 givenname: Patrick S surname: Lee fullname: Lee, Patrick S organization: Infectious Diseases, Novartis Institutes for Biomedical Research, Emeryville, California, USA – sequence: 10 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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ContentType | Journal Article |
Copyright | Copyright © 2019 American Society for Microbiology. Copyright © 2019 American Society for Microbiology. 2019 American Society for Microbiology |
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DocumentTitleAlternate | Resistance to Efflux Pump Inhibition in , , Ranjitkar et al Resistance to Efflux Pump Inhibition in P. aeruginosa |
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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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Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 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. |
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PublicationTitle | Antimicrobial agents and chemotherapy |
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Publisher | American Society for Microbiology |
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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... |
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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 |
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