Lactonase Specificity Is Key to Quorum Quenching in Pseudomonas aeruginosa

Benjamin Rémy; Laure Plener; Philippe Decloquement; Nicholas Armstrong; Mikael Elias; David Daudé; Éric Chabrière

doi:10.3389/fmicb.2020.00762

Lactonase Specificity Is Key to Quorum Quenching in Pseudomonas aeruginosa

Benjamin Rémy, Laure Plener, Philippe Decloquement, Nicholas Armstrong, Mikael Elias, David Daudé, Éric Chabrière

Synthetic Biology and Biotechnology Division

Research output: Contribution to journal › Article › peer-review

20 Scopus citations

Abstract

The human opportunistic pathogen Pseudomonas aeruginosa orchestrates the expression of many genes in a cell density-dependent manner by using quorum sensing (QS). Two acyl-homoserine lactones (AHLs) are involved in QS circuits and contribute to the regulation of virulence factors production, biofilm formation, and antimicrobial sensitivity. Disrupting QS, a strategy referred to as quorum quenching (QQ) can be achieved using exogenous AHL-degrading lactonases. However, the importance of enzyme specificity on quenching efficacy has been poorly investigated. Here, we used two lactonases both targeting the signal molecules N-(3-oxododecanoyl)-L-homoserine lactone (3-oxo-C₁₂ HSL) and butyryl-homoserine lactone (C₄ HSL) albeit with different efficacies on C₄ HSL. Interestingly, both lactonases similarly decreased AHL concentrations and comparably impacted the expression of AHL-based QS genes. However, strong variations were observed in Pseudomonas Quinolone Signal (PQS) regulation depending on the lactonase used. Both lactonases were also found to decrease virulence factors production and biofilm formation in vitro, albeit with different efficiencies. Unexpectedly, only the lactonase with lower efficacy on C₄ HSL was able to inhibit P. aeruginosa pathogenicity in vivo in an amoeba infection model. Similarly, proteomic analysis revealed large variations in protein levels involved in antibiotic resistance, biofilm formation, virulence and diverse cellular mechanisms depending on the chosen lactonase. This global analysis provides evidences that QQ enzyme specificity has a significant impact on the modulation of QS-associated behavior in P. aeruginosa PA14.

Original language	English (US)
Article number	762
Journal	Frontiers in Microbiology
Volume	11
DOIs	https://doi.org/10.3389/fmicb.2020.00762
State	Published - Apr 24 2020

Bibliographical note

Funding Information:
The authors would like to thank Prof. Bernard Lascola and Lina Barrassi for their advice on the amoeba virulence assay, Dr. Raphael Lami for providing AHL reporter strains and Prof. Sophie Bleves for the E. coli competition assay. The authors also thank Nathan Hoekstra and Sara Maitland for corrections on the manuscript. Funding. This work was granted by a project RAPID (LACTO-TEX) from Direction Générale de l’Armement (DGA, France). This work was also supported by “Investissements d’avenir” program (Méditerranée Infection 10-IAHU-03) of the French Agence Nationale de la Recherche (ANR). BR is a Ph.D. student granted by the “Emplois Jeunes Doctorants” program of Région Provence-Alpes-Côte d’Azur (PACA, France). ME was supported by the MnDrive initiative.

Funding Information:
This work was granted by a project RAPID (LACTO-TEX) from Direction Générale de l’Armement (DGA, France). This work was also supported by “Investissements d’avenir” program (Méditerranée Infection 10-IAHU-03) of the French Agence Nationale de la Recherche (ANR). BR is a Ph.D. student granted by the “Emplois Jeunes Doctorants” program of Région Provence-Alpes-Côte d’Azur (PACA, France). ME was supported by the MnDrive initiative.

Publisher Copyright:
© Copyright © 2020 Rémy, Plener, Decloquement, Armstrong, Elias, Daudé and Chabrière.

Keywords

Pseudomonas aeruginosa
acyl-homoserine lactones
biofilm
lactonases
quorum quenching
quorum sensing
virulence

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10.3389/fmicb.2020.00762

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title = "Lactonase Specificity Is Key to Quorum Quenching in Pseudomonas aeruginosa",

abstract = "The human opportunistic pathogen Pseudomonas aeruginosa orchestrates the expression of many genes in a cell density-dependent manner by using quorum sensing (QS). Two acyl-homoserine lactones (AHLs) are involved in QS circuits and contribute to the regulation of virulence factors production, biofilm formation, and antimicrobial sensitivity. Disrupting QS, a strategy referred to as quorum quenching (QQ) can be achieved using exogenous AHL-degrading lactonases. However, the importance of enzyme specificity on quenching efficacy has been poorly investigated. Here, we used two lactonases both targeting the signal molecules N-(3-oxododecanoyl)-L-homoserine lactone (3-oxo-C12 HSL) and butyryl-homoserine lactone (C4 HSL) albeit with different efficacies on C4 HSL. Interestingly, both lactonases similarly decreased AHL concentrations and comparably impacted the expression of AHL-based QS genes. However, strong variations were observed in Pseudomonas Quinolone Signal (PQS) regulation depending on the lactonase used. Both lactonases were also found to decrease virulence factors production and biofilm formation in vitro, albeit with different efficiencies. Unexpectedly, only the lactonase with lower efficacy on C4 HSL was able to inhibit P. aeruginosa pathogenicity in vivo in an amoeba infection model. Similarly, proteomic analysis revealed large variations in protein levels involved in antibiotic resistance, biofilm formation, virulence and diverse cellular mechanisms depending on the chosen lactonase. This global analysis provides evidences that QQ enzyme specificity has a significant impact on the modulation of QS-associated behavior in P. aeruginosa PA14.",

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note = "Funding Information: The authors would like to thank Prof. Bernard Lascola and Lina Barrassi for their advice on the amoeba virulence assay, Dr. Raphael Lami for providing AHL reporter strains and Prof. Sophie Bleves for the E. coli competition assay. The authors also thank Nathan Hoekstra and Sara Maitland for corrections on the manuscript. Funding. This work was granted by a project RAPID (LACTO-TEX) from Direction G{\'e}n{\'e}rale de l{\textquoteright}Armement (DGA, France). This work was also supported by “Investissements d{\textquoteright}avenir” program (M{\'e}diterran{\'e}e Infection 10-IAHU-03) of the French Agence Nationale de la Recherche (ANR). BR is a Ph.D. student granted by the “Emplois Jeunes Doctorants” program of R{\'e}gion Provence-Alpes-C{\^o}te d{\textquoteright}Azur (PACA, France). ME was supported by the MnDrive initiative. Funding Information: This work was granted by a project RAPID (LACTO-TEX) from Direction G{\'e}n{\'e}rale de l{\textquoteright}Armement (DGA, France). This work was also supported by “Investissements d{\textquoteright}avenir” program (M{\'e}diterran{\'e}e Infection 10-IAHU-03) of the French Agence Nationale de la Recherche (ANR). BR is a Ph.D. student granted by the “Emplois Jeunes Doctorants” program of R{\'e}gion Provence-Alpes-C{\^o}te d{\textquoteright}Azur (PACA, France). ME was supported by the MnDrive initiative. Publisher Copyright: {\textcopyright} Copyright {\textcopyright} 2020 R{\'e}my, Plener, Decloquement, Armstrong, Elias, Daud{\'e} and Chabri{\`e}re.",

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AU - Decloquement, Philippe

AU - Armstrong, Nicholas

AU - Elias, Mikael

AU - Daudé, David

AU - Chabrière, Éric

N1 - Funding Information: The authors would like to thank Prof. Bernard Lascola and Lina Barrassi for their advice on the amoeba virulence assay, Dr. Raphael Lami for providing AHL reporter strains and Prof. Sophie Bleves for the E. coli competition assay. The authors also thank Nathan Hoekstra and Sara Maitland for corrections on the manuscript. Funding. This work was granted by a project RAPID (LACTO-TEX) from Direction Générale de l’Armement (DGA, France). This work was also supported by “Investissements d’avenir” program (Méditerranée Infection 10-IAHU-03) of the French Agence Nationale de la Recherche (ANR). BR is a Ph.D. student granted by the “Emplois Jeunes Doctorants” program of Région Provence-Alpes-Côte d’Azur (PACA, France). ME was supported by the MnDrive initiative. Funding Information: This work was granted by a project RAPID (LACTO-TEX) from Direction Générale de l’Armement (DGA, France). This work was also supported by “Investissements d’avenir” program (Méditerranée Infection 10-IAHU-03) of the French Agence Nationale de la Recherche (ANR). BR is a Ph.D. student granted by the “Emplois Jeunes Doctorants” program of Région Provence-Alpes-Côte d’Azur (PACA, France). ME was supported by the MnDrive initiative. Publisher Copyright: © Copyright © 2020 Rémy, Plener, Decloquement, Armstrong, Elias, Daudé and Chabrière.

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Y1 - 2020/4/24

N2 - The human opportunistic pathogen Pseudomonas aeruginosa orchestrates the expression of many genes in a cell density-dependent manner by using quorum sensing (QS). Two acyl-homoserine lactones (AHLs) are involved in QS circuits and contribute to the regulation of virulence factors production, biofilm formation, and antimicrobial sensitivity. Disrupting QS, a strategy referred to as quorum quenching (QQ) can be achieved using exogenous AHL-degrading lactonases. However, the importance of enzyme specificity on quenching efficacy has been poorly investigated. Here, we used two lactonases both targeting the signal molecules N-(3-oxododecanoyl)-L-homoserine lactone (3-oxo-C12 HSL) and butyryl-homoserine lactone (C4 HSL) albeit with different efficacies on C4 HSL. Interestingly, both lactonases similarly decreased AHL concentrations and comparably impacted the expression of AHL-based QS genes. However, strong variations were observed in Pseudomonas Quinolone Signal (PQS) regulation depending on the lactonase used. Both lactonases were also found to decrease virulence factors production and biofilm formation in vitro, albeit with different efficiencies. Unexpectedly, only the lactonase with lower efficacy on C4 HSL was able to inhibit P. aeruginosa pathogenicity in vivo in an amoeba infection model. Similarly, proteomic analysis revealed large variations in protein levels involved in antibiotic resistance, biofilm formation, virulence and diverse cellular mechanisms depending on the chosen lactonase. This global analysis provides evidences that QQ enzyme specificity has a significant impact on the modulation of QS-associated behavior in P. aeruginosa PA14.

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KW - lactonases

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Lactonase Specificity Is Key to Quorum Quenching in Pseudomonas aeruginosa

Abstract

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