The <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Lectin LecB Causes Integrin Internalization and Inhibits Epithelial Wound Healing

ABSTRACT The opportunistic bacterium Pseudomonas aeruginosa produces the fucose-specific lectin LecB, which has been identified as a virulence factor. LecB has a tetrameric structure with four opposing binding sites and has been shown to act as a cross-linker. Here, we demonstrate that LecB strongly...

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Autores principales:	Roland Thuenauer, Alessia Landi, Anne Trefzer, Silke Altmann, Sarah Wehrum, Thorsten Eierhoff, Britta Diedrich, Jörn Dengjel, Alexander Nyström, Anne Imberty, Winfried Römer
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	epithelial cells fucose glycosylation integrins laminin membrane invagination Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/ac2ce578ce1e4faea374eba74d39930f
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spelling	oai:doaj.org-article:ac2ce578ce1e4faea374eba74d39930f2021-11-15T15:57:01ZThe <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Lectin LecB Causes Integrin Internalization and Inhibits Epithelial Wound Healing10.1128/mBio.03260-192150-7511https://doaj.org/article/ac2ce578ce1e4faea374eba74d39930f2020-04-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.03260-19https://doaj.org/toc/2150-7511ABSTRACT The opportunistic bacterium Pseudomonas aeruginosa produces the fucose-specific lectin LecB, which has been identified as a virulence factor. LecB has a tetrameric structure with four opposing binding sites and has been shown to act as a cross-linker. Here, we demonstrate that LecB strongly binds to the glycosylated moieties of β1-integrins on the basolateral plasma membrane of epithelial cells and causes rapid integrin endocytosis. Whereas internalized integrins were degraded via a lysosomal pathway, washout of LecB restored integrin cell surface localization, thus indicating a specific and direct action of LecB on integrins to bring about their endocytosis. Interestingly, LecB was able to trigger uptake of active and inactive β1-integrins and also of complete α3β1-integrin–laminin complexes. We provide a mechanistic explanation for this unique endocytic process by showing that LecB has the additional ability to recognize fucose-bearing glycosphingolipids and causes the formation of membrane invaginations on giant unilamellar vesicles. In cells, LecB recruited integrins to these invaginations by cross-linking integrins and glycosphingolipids. In epithelial wound healing assays, LecB specifically cleared integrins from the surface of cells located at the wound edge and blocked cell migration and wound healing in a dose-dependent manner. Moreover, the wild-type P. aeruginosa strain PAO1 was able to loosen cell-substrate adhesion in order to crawl underneath exposed cells, whereas knockout of LecB significantly reduced crawling events. Based on these results, we suggest that LecB has a role in disseminating bacteria along the cell-basement membrane interface. IMPORTANCE Pseudomonas aeruginosa is a ubiquitous environmental bacterium that is one of the leading causes of nosocomial infections. P. aeruginosa is able to switch between planktonic, intracellular, and biofilm-based lifestyles, which allows it to evade the immune system as well as antibiotic treatment. Hence, alternatives to antibiotic treatment are urgently required to combat P. aeruginosa infections. Lectins, like the fucose-specific LecB, are promising targets, because removal of LecB resulted in decreased virulence in mouse models. Currently, several research groups are developing LecB inhibitors. However, the role of LecB in host-pathogen interactions is not well understood. The significance of our research is in identifying cellular mechanisms of how LecB facilitates P. aeruginosa infection. We introduce LecB as a new member of the list of bacterial molecules that bind integrins and show that P. aeruginosa can move forward underneath attached epithelial cells by loosening cell-basement membrane attachment in a LecB-dependent manner.Roland ThuenauerAlessia LandiAnne TrefzerSilke AltmannSarah WehrumThorsten EierhoffBritta DiedrichJörn DengjelAlexander NyströmAnne ImbertyWinfried RömerAmerican Society for Microbiologyarticleepithelial cellsfucoseglycosylationintegrinslamininmembrane invaginationMicrobiologyQR1-502ENmBio, Vol 11, Iss 2 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	epithelial cells fucose glycosylation integrins laminin membrane invagination Microbiology QR1-502
spellingShingle	epithelial cells fucose glycosylation integrins laminin membrane invagination Microbiology QR1-502 Roland Thuenauer Alessia Landi Anne Trefzer Silke Altmann Sarah Wehrum Thorsten Eierhoff Britta Diedrich Jörn Dengjel Alexander Nyström Anne Imberty Winfried Römer The <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Lectin LecB Causes Integrin Internalization and Inhibits Epithelial Wound Healing
description	ABSTRACT The opportunistic bacterium Pseudomonas aeruginosa produces the fucose-specific lectin LecB, which has been identified as a virulence factor. LecB has a tetrameric structure with four opposing binding sites and has been shown to act as a cross-linker. Here, we demonstrate that LecB strongly binds to the glycosylated moieties of β1-integrins on the basolateral plasma membrane of epithelial cells and causes rapid integrin endocytosis. Whereas internalized integrins were degraded via a lysosomal pathway, washout of LecB restored integrin cell surface localization, thus indicating a specific and direct action of LecB on integrins to bring about their endocytosis. Interestingly, LecB was able to trigger uptake of active and inactive β1-integrins and also of complete α3β1-integrin–laminin complexes. We provide a mechanistic explanation for this unique endocytic process by showing that LecB has the additional ability to recognize fucose-bearing glycosphingolipids and causes the formation of membrane invaginations on giant unilamellar vesicles. In cells, LecB recruited integrins to these invaginations by cross-linking integrins and glycosphingolipids. In epithelial wound healing assays, LecB specifically cleared integrins from the surface of cells located at the wound edge and blocked cell migration and wound healing in a dose-dependent manner. Moreover, the wild-type P. aeruginosa strain PAO1 was able to loosen cell-substrate adhesion in order to crawl underneath exposed cells, whereas knockout of LecB significantly reduced crawling events. Based on these results, we suggest that LecB has a role in disseminating bacteria along the cell-basement membrane interface. IMPORTANCE Pseudomonas aeruginosa is a ubiquitous environmental bacterium that is one of the leading causes of nosocomial infections. P. aeruginosa is able to switch between planktonic, intracellular, and biofilm-based lifestyles, which allows it to evade the immune system as well as antibiotic treatment. Hence, alternatives to antibiotic treatment are urgently required to combat P. aeruginosa infections. Lectins, like the fucose-specific LecB, are promising targets, because removal of LecB resulted in decreased virulence in mouse models. Currently, several research groups are developing LecB inhibitors. However, the role of LecB in host-pathogen interactions is not well understood. The significance of our research is in identifying cellular mechanisms of how LecB facilitates P. aeruginosa infection. We introduce LecB as a new member of the list of bacterial molecules that bind integrins and show that P. aeruginosa can move forward underneath attached epithelial cells by loosening cell-basement membrane attachment in a LecB-dependent manner.
format	article
author	Roland Thuenauer Alessia Landi Anne Trefzer Silke Altmann Sarah Wehrum Thorsten Eierhoff Britta Diedrich Jörn Dengjel Alexander Nyström Anne Imberty Winfried Römer
author_facet	Roland Thuenauer Alessia Landi Anne Trefzer Silke Altmann Sarah Wehrum Thorsten Eierhoff Britta Diedrich Jörn Dengjel Alexander Nyström Anne Imberty Winfried Römer
author_sort	Roland Thuenauer
title	The <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Lectin LecB Causes Integrin Internalization and Inhibits Epithelial Wound Healing
title_short	The <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Lectin LecB Causes Integrin Internalization and Inhibits Epithelial Wound Healing
title_full	The <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Lectin LecB Causes Integrin Internalization and Inhibits Epithelial Wound Healing
title_fullStr	The <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Lectin LecB Causes Integrin Internalization and Inhibits Epithelial Wound Healing
title_full_unstemmed	The <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Lectin LecB Causes Integrin Internalization and Inhibits Epithelial Wound Healing
title_sort	<named-content content-type="genus-species">pseudomonas aeruginosa</named-content> lectin lecb causes integrin internalization and inhibits epithelial wound healing
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/ac2ce578ce1e4faea374eba74d39930f
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The <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Lectin LecB Causes Integrin Internalization and Inhibits Epithelial Wound Healing

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