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Prevalence of ExoY Activity in Pseudomonas aeruginosa Reference Panel Strains and Impact on Cytotoxicity in Epithelial Cells

ExoY is among the effectors that are injected by the type III secretion system (T3SS) of Pseudomonas aeruginosa into host cells. Inside eukaryotic cells, ExoY interacts with F-actin, which stimulates its potent nucleotidyl cyclase activity to produce cyclic nucleotide monophosphates (cNMPs). ExoY ha...

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Autores principales: Silistre, Hazel, Raoux-Barbot, Dorothée, Mancinelli, Federica, Sangouard, Flora, Dupin, Alice, Belyy, Alexander, Deruelle, Vincent, Renault, Louis, Ladant, Daniel, Touqui, Lhousseine, Mechold, Undine
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8524455/
https://www.ncbi.nlm.nih.gov/pubmed/34675890
http://dx.doi.org/10.3389/fmicb.2021.666097
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author Silistre, Hazel
Raoux-Barbot, Dorothée
Mancinelli, Federica
Sangouard, Flora
Dupin, Alice
Belyy, Alexander
Deruelle, Vincent
Renault, Louis
Ladant, Daniel
Touqui, Lhousseine
Mechold, Undine
author_facet Silistre, Hazel
Raoux-Barbot, Dorothée
Mancinelli, Federica
Sangouard, Flora
Dupin, Alice
Belyy, Alexander
Deruelle, Vincent
Renault, Louis
Ladant, Daniel
Touqui, Lhousseine
Mechold, Undine
author_sort Silistre, Hazel
collection PubMed
description ExoY is among the effectors that are injected by the type III secretion system (T3SS) of Pseudomonas aeruginosa into host cells. Inside eukaryotic cells, ExoY interacts with F-actin, which stimulates its potent nucleotidyl cyclase activity to produce cyclic nucleotide monophosphates (cNMPs). ExoY has broad substrate specificity with GTP as a preferential substrate in vitro. How ExoY contributes to the virulence of P. aeruginosa remains largely unknown. Here, we examined the prevalence of active ExoY among strains from the international P. aeruginosa reference panel, a collection of strains that includes environmental and clinical isolates, commonly used laboratory strains, and sequential clonal isolates from cystic fibrosis (CF) patients and thus represents the large diversity of this bacterial species. The ability to secrete active ExoY was determined by measuring the F-actin stimulated guanylate cyclase (GC) activity in bacterial culture supernatants. We found an overall ExoY activity prevalence of about 60% among the 40 examined strains with no significant difference between CF and non-CF isolates. In parallel, we used cellular infection models of human lung epithelial cells to compare the cytotoxic effects of isogenic reference strains expressing active ExoY or lacking the exoY gene. We found that P. aeruginosa strains lacking ExoY were in fact more cytotoxic to the epithelial cells than those secreting active ExoY. This suggests that under certain conditions, ExoY might partly alleviate the cytotoxic effects of other virulence factors of P. aeruginosa.
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spelling pubmed-85244552021-10-20 Prevalence of ExoY Activity in Pseudomonas aeruginosa Reference Panel Strains and Impact on Cytotoxicity in Epithelial Cells Silistre, Hazel Raoux-Barbot, Dorothée Mancinelli, Federica Sangouard, Flora Dupin, Alice Belyy, Alexander Deruelle, Vincent Renault, Louis Ladant, Daniel Touqui, Lhousseine Mechold, Undine Front Microbiol Microbiology ExoY is among the effectors that are injected by the type III secretion system (T3SS) of Pseudomonas aeruginosa into host cells. Inside eukaryotic cells, ExoY interacts with F-actin, which stimulates its potent nucleotidyl cyclase activity to produce cyclic nucleotide monophosphates (cNMPs). ExoY has broad substrate specificity with GTP as a preferential substrate in vitro. How ExoY contributes to the virulence of P. aeruginosa remains largely unknown. Here, we examined the prevalence of active ExoY among strains from the international P. aeruginosa reference panel, a collection of strains that includes environmental and clinical isolates, commonly used laboratory strains, and sequential clonal isolates from cystic fibrosis (CF) patients and thus represents the large diversity of this bacterial species. The ability to secrete active ExoY was determined by measuring the F-actin stimulated guanylate cyclase (GC) activity in bacterial culture supernatants. We found an overall ExoY activity prevalence of about 60% among the 40 examined strains with no significant difference between CF and non-CF isolates. In parallel, we used cellular infection models of human lung epithelial cells to compare the cytotoxic effects of isogenic reference strains expressing active ExoY or lacking the exoY gene. We found that P. aeruginosa strains lacking ExoY were in fact more cytotoxic to the epithelial cells than those secreting active ExoY. This suggests that under certain conditions, ExoY might partly alleviate the cytotoxic effects of other virulence factors of P. aeruginosa. Frontiers Media S.A. 2021-10-04 /pmc/articles/PMC8524455/ /pubmed/34675890 http://dx.doi.org/10.3389/fmicb.2021.666097 Text en Copyright © 2021 Silistre, Raoux-Barbot, Mancinelli, Sangouard, Dupin, Belyy, Deruelle, Renault, Ladant, Touqui and Mechold. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Silistre, Hazel
Raoux-Barbot, Dorothée
Mancinelli, Federica
Sangouard, Flora
Dupin, Alice
Belyy, Alexander
Deruelle, Vincent
Renault, Louis
Ladant, Daniel
Touqui, Lhousseine
Mechold, Undine
Prevalence of ExoY Activity in Pseudomonas aeruginosa Reference Panel Strains and Impact on Cytotoxicity in Epithelial Cells
title Prevalence of ExoY Activity in Pseudomonas aeruginosa Reference Panel Strains and Impact on Cytotoxicity in Epithelial Cells
title_full Prevalence of ExoY Activity in Pseudomonas aeruginosa Reference Panel Strains and Impact on Cytotoxicity in Epithelial Cells
title_fullStr Prevalence of ExoY Activity in Pseudomonas aeruginosa Reference Panel Strains and Impact on Cytotoxicity in Epithelial Cells
title_full_unstemmed Prevalence of ExoY Activity in Pseudomonas aeruginosa Reference Panel Strains and Impact on Cytotoxicity in Epithelial Cells
title_short Prevalence of ExoY Activity in Pseudomonas aeruginosa Reference Panel Strains and Impact on Cytotoxicity in Epithelial Cells
title_sort prevalence of exoy activity in pseudomonas aeruginosa reference panel strains and impact on cytotoxicity in epithelial cells
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8524455/
https://www.ncbi.nlm.nih.gov/pubmed/34675890
http://dx.doi.org/10.3389/fmicb.2021.666097
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