title: Plasma membrane targeting of podocin through the classical exocytic pathway: effect of NPHS2 mutations.
abstract: Podocytes are specialized epithelial cells of the glomerulus in the kidney, which interconnect at the top of the glomerular basement membrane through the slit diaphragm, an adherens-like junction that plays a crucial role in the glomerular filtration process. Podocin, a plasma membrane anchored stomatin-like protein, is expressed in lipid rafts at the insertion of the slit diaphragm in podocytes. Mutations in NPHS2, the gene encoding podocin, are associated with inherited and sporadic cases of steroid-resistant nephrotic syndrome. Here, we show that brefeldin A induces accumulation of newly synthesized podocin in the endoplasmic reticulum, suggesting that podocin biosynthesis follows the classical secretory pathway, and we study the effect of 12 NPHS2 mutations associated with steroid-resistant nephrotic syndrome on the trafficking of the protein. We found that 9 podocin mutants were not targeted to the plasma membrane, 8 being retained in the endoplasmic reticulum and one being localized in late endosomes. Furthermore, by screening our database of patients with NPHS2 mutations, we found that podocin mutants retained in the endoplasmic reticulum are associated with earlier onset of the disease than those correctly targeted to the cell membrane. Our data suggest that most of NPHS2 mutations lead to retention of podocin in the endoplasmic reticulum and therefore provide a rationale for devising therapeutic approaches aimed at correcting the protein processing defect.
authors.affiliation: Inserm U574, Necker Hospital, University Paris 5, Paris, France.
authors.forename: Séverine
authors.initials: S
authors.lastname: Roselli
authors.forename: Imane
authors.initials: I
authors.lastname: Moutkine
authors.forename: Olivier
authors.initials: O
authors.lastname: Gribouval
authors.forename: Alexandre
authors.initials: A
authors.lastname: Benmerah
authors.forename: Corinne
authors.initials: C
authors.lastname: Antignac
mesh_terms: D000818:Animals; D020126:Brefeldin A / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005089:Exocytosis / Q000502:physiology*; D005838:Genotype; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism*; D009154:Mutation; D009404:Nephrotic Syndrome / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D011500:Protein Synthesis Inhibitors / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C407223:NPHS2 protein; D011500:Protein Synthesis Inhibitors; D020126:Brefeldin A