title: Phosphatidylinositol 3-kinase interacts with the glucocorticoid receptor upon TLR2 activation.
abstract: Airway inflammation is a common condition where glucocorticoids (GC) are a well-established therapy. It has been demonstrated that GC stimulate components of innate immunity. Specifically, GC up-regulate TLR2 expression and activation upon inflammatory stimuli; however, little is known about the signalling involved in this process. To determine the mechanism by which dexamethasone modulates TLR2-induced cytokine production this signalling pathway was monitored in a lung epithelial cell line exposed to the TLR2 synthetic agonist, Pam(3) -Cys-Ser-Lys(4) . These experiments demonstrate that phosphatidylinositol 3-kinase (PI3K) is critical for the TLR2 downstream effects of GC. Cells expressing a PI3K mutant (p85-dominant negative, DN; p85 Δ478-511) and exposed to Pam(3) -Cys-Ser-Lys(4)  in the presence or absence of dexamethasone, showed enhanced tumour necrosis factor (TNF)α expression while AP-1 and NF-κB transcriptional activity were repressed. We provide experimental evidence that PI3K physically interacts with the glucocorticoid receptor (GR) through two putative PI3K recruitment consensus YxxM binding motifs in the GR, suggesting that some functions regulated by this receptor might occur through kinase interaction. Mutations of two tyrosine residues in the GR, 598 and 663, to phenylalanine significantly reduced interaction with PI3K and the GC effects on TLR2-induced TNF-α expression. However, these mutations did not alter GR transcriptional activity nor affect cellular localization of the expressed mutant GR in COS-1 cells. Therefore, the PI3K-GR interaction may contribute to the effects of GC on the TLR2 pro-inflammatory signalling cascade, thus defining a novel signalling mechanism with a profound impact on innate immune responses.
authors.affiliation: Immunology Disciplinary Program, Biomedical Sciences Institute, School of Medicine, University of Chile, Santiago, Chile.
authors.forename: Sergio
authors.initials: S
authors.lastname: Arancibia
authors.forename: Dixán
authors.initials: D
authors.lastname: Benítez
authors.forename: Lucia E
authors.initials: LE
authors.lastname: Núñez
authors.forename: Christine M
authors.initials: CM
authors.lastname: Jewell
authors.forename: Patricia
authors.initials: P
authors.lastname: Langjahr
authors.forename: Enzo
authors.initials: E
authors.lastname: Candia
authors.forename: Gerald
authors.initials: G
authors.lastname: Zapata-Torres
authors.forename: John A
authors.initials: JA
authors.lastname: Cidlowski
authors.forename: María-Julieta
authors.initials: MJ
authors.lastname: González
authors.forename: Marcela A
authors.initials: MA
authors.lastname: Hermoso
mesh_terms: D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis; D003907:Dexamethasone / Q000494:pharmacology*; D004847:Epithelial Cells / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D057809:HEK293 Cells; D006801:Humans; D008168:Lung / Q000378:metabolism; D016328:NF-kappa B / Q000096:biosynthesis; D010455:Peptides / Q000494:pharmacology; D058539:Phosphatidylinositol 3-Kinase / Q000235:genetics / Q000378:metabolism*; D011965:Receptors, Glucocorticoid / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D051195:Toll-Like Receptor 2 / Q000276:immunology* / Q000378:metabolism; D018808:Transcription Factor AP-1 / Q000096:biosynthesis; D015533:Transcriptional Activation; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000378:metabolism
chemical_list: D016207:Cytokines; D016328:NF-kappa B; D010455:Peptides; D011965:Receptors, Glucocorticoid; C495342:TLR2 protein, human; D051195:Toll-Like Receptor 2; D018808:Transcription Factor AP-1; D014409:Tumor Necrosis Factor-alpha; D003907:Dexamethasone; D058539:Phosphatidylinositol 3-Kinase