A Small Multidrug Resistance Transporter in<i>Pseudomonas aeruginosa</i>Confers Substrate-Specific Resistance or Susceptibility
ABSTRACT Small Multidrug Resistance (SMR) transporters are key players in the defense of multidrug-resistant pathogens to toxins and other homeostasis-perturbing compounds. However, recent evidence demonstrates that EmrE, an SMR fromEscherichia coliand a model for understanding transport, can also induce susceptibility to some compounds by drug-gated proton leak. This runs down the ΔpH component of the Proton Motive Force (PMF), reducing viability of the affected bacteria. Proton leak may provide an unexplored drug target distinct from the targets of most known antibiotics. Activating proton leak requires an SMR to be merely present, rather than be the primary resistance mechanism, and dissipates the energy source for many other efflux pumps. PAsmr, an EmrE homolog fromP. aeruginosa, transports many EmrE substrates in cells and purified systems. We hypothesized that PAsmr, like EmrE, may confer susceptibility to some compounds via drug-gated proton leak. Growth assays ofE. coliexpressing PAsmr displayed substrate-dependent resistance and susceptibility phenotypes, andin vitrosolid-supported membrane electrophysiology experiments revealed that PAsmr performs both antiport and substrate-gated proton uniport, demonstrating the same functional promiscuity observed in EmrE. Growth assays ofP. aeruginosastrain PA14 demonstrated that PAsmr contributes resistance to some antimicrobial compounds, but no growth defect is observed with susceptibility substrates, suggestingP. aeruginosacan compensate for the proton leak occurring through PAsmr. These phenotypic differences betweenP. aeruginosaandE. coliadvance our understanding of underlying resistance mechanisms inP. aeruginosaand prompt further investigation into the role that SMRs play in antibiotic resistance in pathogens.IMPORTANCE Small multidrug resistance transporters are a class of efflux pumps found in many pathogens, but whose contributions to antibiotic resistance are not fully understood. We hypothesize that these transporters may confer not only resistance, but also susceptibility, by dissipating the proton-motive force. This means to use an SMR transporter as a target, it merely needs to be present (as opposed to being the primary resistance mechanism). Here, we test this hypothesis with an SMR transporter found inPseudomonas aeruginosaand find that it can perform both antiport (conferring resistance) and substrate-gated proton leak. Proton leak is detrimental to growth inE. colibut notP. aeruginosa, suggesting thatP. aeruginosaresponds differently to or can altogether prevent ΔpH dissipation..
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Preprint| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
bioRxiv.org - (2024) vom: 16. Apr. Zur Gesamtaufnahme - year:2024 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
Wegrzynowicz, Andrea K. [VerfasserIn] |
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Volltext [kostenfrei] |
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| doi: |
10.1101/2023.09.28.560013 |
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XBI040990249 |
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| 520 | |a ABSTRACT Small Multidrug Resistance (SMR) transporters are key players in the defense of multidrug-resistant pathogens to toxins and other homeostasis-perturbing compounds. However, recent evidence demonstrates that EmrE, an SMR fromEscherichia coliand a model for understanding transport, can also induce susceptibility to some compounds by drug-gated proton leak. This runs down the ΔpH component of the Proton Motive Force (PMF), reducing viability of the affected bacteria. Proton leak may provide an unexplored drug target distinct from the targets of most known antibiotics. Activating proton leak requires an SMR to be merely present, rather than be the primary resistance mechanism, and dissipates the energy source for many other efflux pumps. PAsmr, an EmrE homolog fromP. aeruginosa, transports many EmrE substrates in cells and purified systems. We hypothesized that PAsmr, like EmrE, may confer susceptibility to some compounds via drug-gated proton leak. Growth assays ofE. coliexpressing PAsmr displayed substrate-dependent resistance and susceptibility phenotypes, andin vitrosolid-supported membrane electrophysiology experiments revealed that PAsmr performs both antiport and substrate-gated proton uniport, demonstrating the same functional promiscuity observed in EmrE. Growth assays ofP. aeruginosastrain PA14 demonstrated that PAsmr contributes resistance to some antimicrobial compounds, but no growth defect is observed with susceptibility substrates, suggestingP. aeruginosacan compensate for the proton leak occurring through PAsmr. These phenotypic differences betweenP. aeruginosaandE. coliadvance our understanding of underlying resistance mechanisms inP. aeruginosaand prompt further investigation into the role that SMRs play in antibiotic resistance in pathogens.IMPORTANCE Small multidrug resistance transporters are a class of efflux pumps found in many pathogens, but whose contributions to antibiotic resistance are not fully understood. We hypothesize that these transporters may confer not only resistance, but also susceptibility, by dissipating the proton-motive force. This means to use an SMR transporter as a target, it merely needs to be present (as opposed to being the primary resistance mechanism). Here, we test this hypothesis with an SMR transporter found inPseudomonas aeruginosaand find that it can perform both antiport (conferring resistance) and substrate-gated proton leak. Proton leak is detrimental to growth inE. colibut notP. aeruginosa, suggesting thatP. aeruginosaresponds differently to or can altogether prevent ΔpH dissipation. | ||
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| 700 | 1 | |a Heelan, William J. |e verfasserin |4 aut | |
| 700 | 1 | |a Demas, Sydnye P. |e verfasserin |4 aut | |
| 700 | 1 | |a McLean, Maxwell S. |e verfasserin |4 aut | |
| 700 | 1 | |a Peters, Jason M. |e verfasserin |4 aut | |
| 700 | 1 | |a Henzler-Wildman, Katherine A. |e verfasserin |4 aut | |
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