title: The serine/threonine kinase Par1b regulates epithelial lumen polarity via IRSp53-mediated cell-ECM signaling.
abstract: The serine/threonine kinase Par1b promotes cell-cell adhesion and determines the polarity of the luminal domain in epithelial cells. In this study, we demonstrate that Par1b also regulates cell-extracellular matrix (ECM) signaling in kidney-derived Madin-Darby canine kidney (MDCK) cells and identified the rho-guanosine triphosphatase adaptor and scaffolding protein IRSp53 as a Par1b substrate involved in this pathway. Par1b overexpression inhibits basal lamina formation, cell spreading, focal adhesion, stress fiber formation, and compaction, whereas Par1b depletion has the opposite effect. IRSp53 depletion mimics Par1b overexpression on cell-ECM signaling and lumen polarity but had no effect on adherens junction formation. Par1b directly phosphorylates IRSp53 on S366 in cell lysates and stimulates phosphorylation on S453/3/5 via an indirect mechanism. A Par1b phosphorylation-deficient IRSp53 mutant but not the wild-type protein efficiently rescues both the cell spreading and the lumen polarity defects in Par1b MDCK cells. Our data suggest a model in which Par1b phosphorylation prevents recruitment of IRSp53 effector proteins to its Src homology domain 3 by promoting 14-3-3 binding in the vicinity of that domain.
authors.affiliation: Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, The Bronx, NY 10461, USA.
authors.forename: David
authors.initials: D
authors.lastname: Cohen
authors.forename: Dawn
authors.initials: D
authors.lastname: Fernandez
authors.forename: Francisco
authors.initials: F
authors.lastname: Lázaro-Diéguez
authors.forename: Anne
authors.initials: A
authors.lastname: Müsch
mesh_terms: D048948:14-3-3 Proteins / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion; D016764:Cell Polarity; D002478:Cells, Cultured; D004285:Dogs; D004847:Epithelial Cells / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D018909:src Homology Domains
chemical_list: D048948:14-3-3 Proteins; D009419:Nerve Tissue Proteins; D017346:Protein Serine-Threonine Kinases