title: Phosphoinositide 3-kinase regulates the role of retromer in transcytosis of the polymeric immunoglobulin receptor.
abstract: Retromer is a multimeric protein complex that mediates intracellular receptor sorting. One of the roles of retromer is to promote transcytosis of the polymeric immunoglobulin receptor (pIgR) and its ligand polymeric immunoglobulin A (pIgA) in polarized epithelial cells. In Madin-Darby Canine Kidney (MDCK) cells, overexpression of Vps35, the retromer subunit key for cargo recognition, restores transcytosis to a pIgR mutant that is normally degraded. Here we show that pIgA transcytosis was not restored in these cells when treated with the specific phosphoinositide 3-kinase (PI3K) inhibitor LY294002. Likewise, the decrease in pIgA transcytosis by wild-type pIgR seen upon PI3K inhibition was not reverted by Vps35 overexpression. PI3K inhibition reduced membrane association of sorting-nexins (SNX) 1 and 2, which constitute the retromer subcomplex involved in membrane deformation, while association of the Vps35-Vps26-Vps29 subcomplex, involved in cargo recognition, remained virtually unaffected. Colocalization between the two retromer subcomplexes was reduced upon the treatment. Whereas the interaction among the subunits of the Vps35-Vps26-Vps29 subcomplex remained unchanged, less Vps35 was found associated with pIgR upon PI3K inhibition. In addition, colocalization of internalized pIgA with subunits of both retromer subcomplexes throughout the transcytotic pathway was substantially reduced by LY294002 treatment. These data implicate PI3K in controlling retromer's role in pIgR-pIgA transcytosis.
authors.affiliation: Department of Anatomy, Cardiovascular Research Institute and Department of Biochemistry and Biophysics, University of California School of Medicine, San Francisco, California 94143-2140, USA. mverges@cipf.es <mverges@cipf.es>
authors.forename: Marcel
authors.initials: M
authors.lastname: Vergés
authors.forename: Isabel
authors.initials: I
authors.lastname: Sebastián
authors.forename: Keith E
authors.initials: KE
authors.lastname: Mostov
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000502:physiology*; D002478:Cells, Cultured; D002867:Chromones / Q000494:pharmacology; D004285:Dogs; D007070:Immunoglobulin A / Q000378:metabolism; D009025:Morpholines / Q000494:pharmacology; D046912:Multiprotein Complexes / Q000737:chemistry / Q000502:physiology*; D050505:Mutant Proteins / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000502:physiology*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D021381:Protein Transport / Q000187:drug effects; D019056:Receptors, Polymeric Immunoglobulin / Q000378:metabolism*; D014162:Transfection; D033921:Vesicular Transport Proteins / Q000235:genetics / Q000502:physiology*
chemical_list: D002352:Carrier Proteins; D002867:Chromones; D007070:Immunoglobulin A; D009025:Morpholines; D046912:Multiprotein Complexes; D050505:Mutant Proteins; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D019056:Receptors, Polymeric Immunoglobulin; D033921:Vesicular Transport Proteins; C498296:VPS26A protein, human; C419572:VPS29 protein, human; C419573:VPS35 protein, human; C034639:polymeric IgA; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one