<named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>-Induced Bleb-Niche Formation in Epithelial Cells Is Independent of Actinomyosin Contraction and Enhanced by Loss of Cystic Fibrosis Transmembrane-Conductance Regulator Osmoregulatory Function

ABSTRACT The opportunistic pathogen Pseudomonas aeruginosa can infect almost any site in the body but most often targets epithelial cell-lined tissues such as the airways, skin, and the cornea of the eye. A common predisposing factor is cystic fibrosis (CF), caused by defects in the cystic fibrosis...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Amber L. Jolly, Desire Takawira, Olufolarin O. Oke, Sarah A. Whiteside, Stephanie W. Chang, Emily R. Wen, Kevin Quach, David J. Evans, Suzanne M. J. Fleiszig
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2015
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/2b2cd1a5e9ea418786558b7544a2d645
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:2b2cd1a5e9ea418786558b7544a2d645
record_format	dspace
spelling	oai:doaj.org-article:2b2cd1a5e9ea418786558b7544a2d6452021-11-15T15:41:33Z<named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>-Induced Bleb-Niche Formation in Epithelial Cells Is Independent of Actinomyosin Contraction and Enhanced by Loss of Cystic Fibrosis Transmembrane-Conductance Regulator Osmoregulatory Function10.1128/mBio.02533-142150-7511https://doaj.org/article/2b2cd1a5e9ea418786558b7544a2d6452015-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02533-14https://doaj.org/toc/2150-7511ABSTRACT The opportunistic pathogen Pseudomonas aeruginosa can infect almost any site in the body but most often targets epithelial cell-lined tissues such as the airways, skin, and the cornea of the eye. A common predisposing factor is cystic fibrosis (CF), caused by defects in the cystic fibrosis transmembrane-conductance regulator (CFTR). Previously, we showed that when P. aeruginosa enters epithelial cells it replicates intracellularly and occupies plasma membrane blebs. This phenotype is dependent on the type 3 secretion system (T3SS) effector ExoS, shown by others to induce host cell apoptosis. Here, we examined mechanisms for P. aeruginosa-induced bleb formation, focusing on its relationship to apoptosis and the CFTR. The data showed that P. aeruginosa-induced blebbing in epithelial cells is independent of actin contraction and is inhibited by hyperosmotic media (400 to 600 mOsM), distinguishing bacterially induced blebs from apoptotic blebs. Cells with defective CFTR displayed enhanced bleb formation upon infection, as demonstrated using bronchial epithelial cells from a patient with cystic fibrosis and a CFTR inhibitor, CFTR(Inh)-172. The defect was found to be correctable either by incubation in hyperosmotic media or by complementation with CFTR (pGFP-CFTR), suggesting that the osmoregulatory function of CFTR counters P. aeruginosa-induced bleb-niche formation. Accordingly, and despite their reduced capacity for bacterial internalization, CFTR-deficient cells showed greater bacterial occupation of blebs and enhanced intracellular replication. Together, these data suggest that P. aeruginosa bleb niches are distinct from apoptotic blebs, are driven by osmotic forces countered by CFTR, and could provide a novel mechanism for bacterial persistence in the host. IMPORTANCE Pseudomonas aeruginosa is an opportunistic pathogen problematic in hospitalized patients and those with cystic fibrosis (CF). Previously, we showed that P. aeruginosa can enter epithelial cells and replicate within them and traffics to the membrane blebs that it induces. This “bleb-niche” formation requires ExoS, previously shown to cause apoptosis. Here, we show that the driving force for bleb-niche formation is osmotic pressure, differentiating P. aeruginosa-induced blebs from apoptotic blebs. Either CFTR inhibition or CFTR mutation (as seen in people with CF) causes P. aeruginosa to make more bleb niches and provides an osmotic driving force for blebbing. CFTR inhibition also enhances bacterial occupation of blebs and intracellular replication. Since CFTR is targeted for removal from the plasma membrane when P. aeruginosa invades a healthy cell, these findings could relate to pathogenesis in both CF and healthy patient populations.Amber L. JollyDesire TakawiraOlufolarin O. OkeSarah A. WhitesideStephanie W. ChangEmily R. WenKevin QuachDavid J. EvansSuzanne M. J. FleiszigAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 6, Iss 2 (2015)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Amber L. Jolly Desire Takawira Olufolarin O. Oke Sarah A. Whiteside Stephanie W. Chang Emily R. Wen Kevin Quach David J. Evans Suzanne M. J. Fleiszig <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>-Induced Bleb-Niche Formation in Epithelial Cells Is Independent of Actinomyosin Contraction and Enhanced by Loss of Cystic Fibrosis Transmembrane-Conductance Regulator Osmoregulatory Function
description	ABSTRACT The opportunistic pathogen Pseudomonas aeruginosa can infect almost any site in the body but most often targets epithelial cell-lined tissues such as the airways, skin, and the cornea of the eye. A common predisposing factor is cystic fibrosis (CF), caused by defects in the cystic fibrosis transmembrane-conductance regulator (CFTR). Previously, we showed that when P. aeruginosa enters epithelial cells it replicates intracellularly and occupies plasma membrane blebs. This phenotype is dependent on the type 3 secretion system (T3SS) effector ExoS, shown by others to induce host cell apoptosis. Here, we examined mechanisms for P. aeruginosa-induced bleb formation, focusing on its relationship to apoptosis and the CFTR. The data showed that P. aeruginosa-induced blebbing in epithelial cells is independent of actin contraction and is inhibited by hyperosmotic media (400 to 600 mOsM), distinguishing bacterially induced blebs from apoptotic blebs. Cells with defective CFTR displayed enhanced bleb formation upon infection, as demonstrated using bronchial epithelial cells from a patient with cystic fibrosis and a CFTR inhibitor, CFTR(Inh)-172. The defect was found to be correctable either by incubation in hyperosmotic media or by complementation with CFTR (pGFP-CFTR), suggesting that the osmoregulatory function of CFTR counters P. aeruginosa-induced bleb-niche formation. Accordingly, and despite their reduced capacity for bacterial internalization, CFTR-deficient cells showed greater bacterial occupation of blebs and enhanced intracellular replication. Together, these data suggest that P. aeruginosa bleb niches are distinct from apoptotic blebs, are driven by osmotic forces countered by CFTR, and could provide a novel mechanism for bacterial persistence in the host. IMPORTANCE Pseudomonas aeruginosa is an opportunistic pathogen problematic in hospitalized patients and those with cystic fibrosis (CF). Previously, we showed that P. aeruginosa can enter epithelial cells and replicate within them and traffics to the membrane blebs that it induces. This “bleb-niche” formation requires ExoS, previously shown to cause apoptosis. Here, we show that the driving force for bleb-niche formation is osmotic pressure, differentiating P. aeruginosa-induced blebs from apoptotic blebs. Either CFTR inhibition or CFTR mutation (as seen in people with CF) causes P. aeruginosa to make more bleb niches and provides an osmotic driving force for blebbing. CFTR inhibition also enhances bacterial occupation of blebs and intracellular replication. Since CFTR is targeted for removal from the plasma membrane when P. aeruginosa invades a healthy cell, these findings could relate to pathogenesis in both CF and healthy patient populations.
format	article
author	Amber L. Jolly Desire Takawira Olufolarin O. Oke Sarah A. Whiteside Stephanie W. Chang Emily R. Wen Kevin Quach David J. Evans Suzanne M. J. Fleiszig
author_facet	Amber L. Jolly Desire Takawira Olufolarin O. Oke Sarah A. Whiteside Stephanie W. Chang Emily R. Wen Kevin Quach David J. Evans Suzanne M. J. Fleiszig
author_sort	Amber L. Jolly
title	<named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>-Induced Bleb-Niche Formation in Epithelial Cells Is Independent of Actinomyosin Contraction and Enhanced by Loss of Cystic Fibrosis Transmembrane-Conductance Regulator Osmoregulatory Function
title_short	<named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>-Induced Bleb-Niche Formation in Epithelial Cells Is Independent of Actinomyosin Contraction and Enhanced by Loss of Cystic Fibrosis Transmembrane-Conductance Regulator Osmoregulatory Function
title_full	<named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>-Induced Bleb-Niche Formation in Epithelial Cells Is Independent of Actinomyosin Contraction and Enhanced by Loss of Cystic Fibrosis Transmembrane-Conductance Regulator Osmoregulatory Function
title_fullStr	<named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>-Induced Bleb-Niche Formation in Epithelial Cells Is Independent of Actinomyosin Contraction and Enhanced by Loss of Cystic Fibrosis Transmembrane-Conductance Regulator Osmoregulatory Function
title_full_unstemmed	<named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>-Induced Bleb-Niche Formation in Epithelial Cells Is Independent of Actinomyosin Contraction and Enhanced by Loss of Cystic Fibrosis Transmembrane-Conductance Regulator Osmoregulatory Function
title_sort	<named-content content-type="genus-species">pseudomonas aeruginosa</named-content>-induced bleb-niche formation in epithelial cells is independent of actinomyosin contraction and enhanced by loss of cystic fibrosis transmembrane-conductance regulator osmoregulatory function
publisher	American Society for Microbiology
publishDate	2015
url	https://doaj.org/article/2b2cd1a5e9ea418786558b7544a2d645
work_keys_str_mv	AT amberljolly namedcontentcontenttypegenusspeciespseudomonasaeruginosanamedcontentinducedblebnicheformationinepithelialcellsisindependentofactinomyosincontractionandenhancedbylossofcysticfibrosistransmembraneconductanceregulatorosmoregulatoryfunction AT desiretakawira namedcontentcontenttypegenusspeciespseudomonasaeruginosanamedcontentinducedblebnicheformationinepithelialcellsisindependentofactinomyosincontractionandenhancedbylossofcysticfibrosistransmembraneconductanceregulatorosmoregulatoryfunction AT olufolarinooke namedcontentcontenttypegenusspeciespseudomonasaeruginosanamedcontentinducedblebnicheformationinepithelialcellsisindependentofactinomyosincontractionandenhancedbylossofcysticfibrosistransmembraneconductanceregulatorosmoregulatoryfunction AT sarahawhiteside namedcontentcontenttypegenusspeciespseudomonasaeruginosanamedcontentinducedblebnicheformationinepithelialcellsisindependentofactinomyosincontractionandenhancedbylossofcysticfibrosistransmembraneconductanceregulatorosmoregulatoryfunction AT stephaniewchang namedcontentcontenttypegenusspeciespseudomonasaeruginosanamedcontentinducedblebnicheformationinepithelialcellsisindependentofactinomyosincontractionandenhancedbylossofcysticfibrosistransmembraneconductanceregulatorosmoregulatoryfunction AT emilyrwen namedcontentcontenttypegenusspeciespseudomonasaeruginosanamedcontentinducedblebnicheformationinepithelialcellsisindependentofactinomyosincontractionandenhancedbylossofcysticfibrosistransmembraneconductanceregulatorosmoregulatoryfunction AT kevinquach namedcontentcontenttypegenusspeciespseudomonasaeruginosanamedcontentinducedblebnicheformationinepithelialcellsisindependentofactinomyosincontractionandenhancedbylossofcysticfibrosistransmembraneconductanceregulatorosmoregulatoryfunction AT davidjevans namedcontentcontenttypegenusspeciespseudomonasaeruginosanamedcontentinducedblebnicheformationinepithelialcellsisindependentofactinomyosincontractionandenhancedbylossofcysticfibrosistransmembraneconductanceregulatorosmoregulatoryfunction AT suzannemjfleiszig namedcontentcontenttypegenusspeciespseudomonasaeruginosanamedcontentinducedblebnicheformationinepithelialcellsisindependentofactinomyosincontractionandenhancedbylossofcysticfibrosistransmembraneconductanceregulatorosmoregulatoryfunction
_version_	1718427633131192320

<named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>-Induced Bleb-Niche Formation in Epithelial Cells Is Independent of Actinomyosin Contraction and Enhanced by Loss of Cystic Fibrosis Transmembrane-Conductance Regulator Osmoregulatory Function

Ejemplares similares