title: A role of OCRL in clathrin-coated pit dynamics and uncoating revealed by studies of Lowe syndrome cells.
abstract: Mutations in the inositol 5-phosphatase OCRL cause Lowe syndrome and Dent's disease. Although OCRL, a direct clathrin interactor, is recruited to late-stage clathrin-coated pits, clinical manifestations have been primarily attributed to intracellular sorting defects. Here we show that OCRL loss in Lowe syndrome patient fibroblasts impacts clathrin-mediated endocytosis and results in an endocytic defect. These cells exhibit an accumulation of clathrin-coated vesicles and an increase in U-shaped clathrin-coated pits, which may result from sequestration of coat components on uncoated vesicles. Endocytic vesicles that fail to lose their coat nucleate the majority of the numerous actin comets present in patient cells. SNX9, an adaptor that couples late-stage endocytic coated pits to actin polymerization and which we found to bind OCRL directly, remains associated with such vesicles. These results indicate that OCRL acts as an uncoating factor and that defects in clathrin-mediated endocytosis likely contribute to pathology in patients with OCRL mutations.
authors.affiliation: Department of Cell Biology, Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United States.
authors.forename: Ramiro
authors.initials: R
authors.lastname: Nández
authors.affiliation: Department of Cell Biology, Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United States.
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Balkin
authors.affiliation: Department of Cell Biology, Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United States.
authors.forename: Mirko
authors.initials: M
authors.lastname: Messa
authors.affiliation: Department of Diagnostic Radiology, Yale University School of Medicine, New Haven, United States.
authors.forename: Liang
authors.initials: L
authors.lastname: Liang
authors.affiliation: Department of Cell Biology, Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United States.
authors.forename: Summer
authors.initials: S
authors.lastname: Paradise
authors.affiliation: Department of Cell Biology, Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United States.
authors.forename: Heather
authors.initials: H
authors.lastname: Czapla
authors.affiliation: Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany.
authors.forename: Marco Y
authors.identifier: http://orcid.org/0000-0002-9490-2261
authors.initials: MY
authors.lastname: Hein
authors.affiliation: Department of Diagnostic Radiology, Yale University School of Medicine, New Haven, United States.
authors.forename: James S
authors.initials: JS
authors.lastname: Duncan
authors.affiliation: Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Mann
authors.affiliation: Department of Cell Biology, Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United States.
authors.forename: Pietro
authors.initials: P
authors.lastname: De Camilli
mesh_terms: D002478:Cells, Cultured; D002966:Clathrin / Q000378:metabolism*; D022163:Clathrin-Coated Vesicles / Q000378:metabolism / Q000648:ultrastructure; D003034:Coated Pits, Cell-Membrane / Q000378:metabolism* / Q000648:ultrastructure; D004705:Endocytosis / Q000235:genetics; D005347:Fibroblasts / Q000378:metabolism*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence / Q000379:methods; D009154:Mutation; D009800:Oculocerebrorenal Syndrome / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D020543:Proteome / Q000235:genetics / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D034622:RNA Interference; D058305:Sorting Nexins / Q000235:genetics / Q000378:metabolism
chemical_list: D002966:Clathrin; D008164:Luminescent Proteins; D018129:Phosphatidylinositol Phosphates; D020543:Proteome; C400533:SNX9 protein, human; D058305:Sorting Nexins; D010744:Phosphoric Monoester Hydrolases; C075695:OCRL protein, human
keywords: Lowe syndrome; PI(4,5)P2; SNX9; actin comets; cell biology; clathrin-mediated endocytosis; human; human biology; medicine; uncoating
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