title: The junctional integrity of epithelial cells is modulated by Pseudomonas aeruginosa quorum sensing molecule through phosphorylation-dependent mechanisms.
abstract: In Pseudomonas aeruginosa, cell-cell communication based on acyl-homoserine lactone (HSL) quorum sensing molecules is known to coordinate the production of virulence factors and biofilms by the bacterium. Incidentally, these bacterial signals can also modulate mammalian cell behaviour. We report that 3O-C(12)-HSL can disrupt adherens junctions in human epithelial Caco-2 cells as evidenced by a reduction of the expression and distribution of E-cadherin and beta-catenin. Using co-immunoprecipitation we also found that P. aeruginosa 3O-C(12)-HSL-treatment resulted in tyrosine hyperphosphorylation of E-cadherin, beta-catenin, occludin and ZO-1. Similarly, serine and threonine residues of E-cadherin and ZO-1 became more phosphorylated after 3O-C(12)-HSL treatment. On the contrary, occludin and beta-catenin underwent dephosphorylation on serine and threonine residues after exposition of 3O-C(12)-HSL. These changes in the phosphorylation state were paralleled by alteration in the structure of junction complexes and increased paracellular permeability. Moreover, pre-treatment of the Caco-2 cells with protein phosphatase and kinase inhibitors prevented 3O-C(12)-HSL-induced changes in paracellular permeability and interactions between occludin-ZO-1 and the E-cadherin-beta-catenin. These findings clearly suggest that an alteration in the phosphorylation status of junction proteins are involved in the changes in cell junction associations and enhanced paracellular permeability, and that bacterial signals are indeed sensed by the host cells.
authors.affiliation: Division of Medical Microbiology, Department of Clinical and Experimental Medicine, Linköping University, SE-58185 Linköping, Sweden. elevi@imk.liu.se
authors.forename: Elena
authors.initials: E
authors.lastname: Vikström
authors.forename: Lan
authors.initials: L
authors.lastname: Bui
authors.forename: Peter
authors.initials: P
authors.lastname: Konradsson
authors.forename: Karl-Eric
authors.initials: KE
authors.lastname: Magnusson
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000494:pharmacology; D018938:Caco-2 Cells; D015820:Cadherins / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D019833:Genistein / Q000494:pharmacology; D006714:Homoserine / Q000031:analogs & derivatives / Q000494:pharmacology; D006801:Humans; D007365:Intercellular Junctions / Q000187:drug effects* / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D062793:Occludin; D019319:Okadaic Acid / Q000494:pharmacology; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D010768:Phosphoserine / Q000378:metabolism; D010769:Phosphothreonine / Q000378:metabolism; D019000:Phosphotyrosine / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D011550:Pseudomonas aeruginosa / Q000737:chemistry*; D053038:Quorum Sensing; D019311:Staurosporine / Q000494:pharmacology; D062826:Zonula Occludens-1 Protein; D051176:beta Catenin / Q000378:metabolism
chemical_list: D015820:Cadherins; D008565:Membrane Proteins; C109860:N-(3-oxododecanoyl)homoserine lactone; C568985:OCLN protein, human; D062793:Occludin; D010750:Phosphoproteins; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein; D051176:beta Catenin; D010769:Phosphothreonine; D010768:Phosphoserine; D019319:Okadaic Acid; D019000:Phosphotyrosine; D006714:Homoserine; D019833:Genistein; D019311:Staurosporine; D015107:4-Butyrolactone