title: Rab35 GTPase Triggers Switch-like Recruitment of the Lowe Syndrome Lipid Phosphatase OCRL on Newborn Endosomes.
abstract: Phosphoinositide (PtdIns) homeostasis requires a tight spatial and temporal regulation during the endocytic process [1]. Indeed, PtdIns(4,5)P2 plays a crucial role in endocytosis by controlling clathrin-coated pit formation, whereas its conversion into PtdIns4P right after scission of clathrin-coated vesicles (CCVs) is essential for successful uncoating and cargo sorting [1-6]. In non-neuronal cells, endosomal PtdIns(4,5)P2 hydrolysis critically relies on the lipid phosphatase OCRL [7-9], the inactivation of which causes the Oculo-Cerebro-Renal syndrome of Lowe [10, 11]. To understand the coupling between PtdIns(4,5)P2 hydrolysis and endosome formation, a key issue is thus to unravel the mechanism by which OCRL is recruited on CCVs precisely after their scission from the plasma membrane. Here we found that the Rab35 GTPase, which plays a fundamental but poorly understood role in endosomal trafficking after cargo internalization [12-21], directly recruits the OCRL phosphatase immediately after scission of the CCVs. Consistent with Rab35 and OCRL acting together, depletion of either Rab35 or OCRL leads to retention of internalized receptors such as the endogenous cation-independent mannose-6-phosphate receptor (CI-MPR) in peripheral clathrin-positive endosomes that display abnormal association with PtdIns(4,5)P2- and actin-binding proteins. Remarkably, Rab35 loading on CCVs rapidly follows the recruitment of the AP2-binding Rab35 GEF/activator DENND1A (connecdenn 1) and the disappearance of the Rab35 GAP/inhibitor EPI64B. We propose that the precise spatial and temporal activation of Rab35 acts as a major switch for OCRL recruitment on newborn endosomes, post-scission PtdIns(4,5)P2 hydrolysis, and subsequent endosomal trafficking.
authors.affiliation: Membrane Traffic and Cell Division Lab, Cell Biology and Infection Department, Institut Pasteur, 25-28 Rue du Dr. Roux, 75724 Paris Cedex 15, France; Centre National de la Recherche Scientifique UMR3691, 75015 Paris, France; Institut de Formation Doctorale, Sorbonne Universités and Université Pierre et Marie Curie, Université Paris 06, 75252 Paris, France.
authors.forename: Clothilde
authors.initials: C
authors.lastname: Cauvin
authors.affiliation: University of Bordeaux, 33000 Bordeaux, France; Centre National de la Recherche Scientifique, Interdisciplinary Institute for Neuroscience, UMR 5297, 33000 Bordeaux, France.
authors.forename: Morgane
authors.initials: M
authors.lastname: Rosendale
authors.affiliation: Membrane Traffic and Cell Division Lab, Cell Biology and Infection Department, Institut Pasteur, 25-28 Rue du Dr. Roux, 75724 Paris Cedex 15, France; Centre National de la Recherche Scientifique UMR3691, 75015 Paris, France.
authors.forename: Neetu
authors.initials: N
authors.lastname: Gupta-Rossi
authors.affiliation: Membrane Traffic and Cell Division Lab, Cell Biology and Infection Department, Institut Pasteur, 25-28 Rue du Dr. Roux, 75724 Paris Cedex 15, France; Centre National de la Recherche Scientifique UMR3691, 75015 Paris, France.
authors.forename: Murielle
authors.initials: M
authors.lastname: Rocancourt
authors.affiliation: Membrane Traffic and Cell Division Lab, Cell Biology and Infection Department, Institut Pasteur, 25-28 Rue du Dr. Roux, 75724 Paris Cedex 15, France; Centre National de la Recherche Scientifique UMR3691, 75015 Paris, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Larraufie
authors.affiliation: Service de Néphrologie Pédiatrique, AP-HP Hôpital Necker, INSERM U983, 75015 Paris, France.
authors.forename: Rémi
authors.initials: R
authors.lastname: Salomon
authors.affiliation: University of Bordeaux, 33000 Bordeaux, France; Centre National de la Recherche Scientifique, Interdisciplinary Institute for Neuroscience, UMR 5297, 33000 Bordeaux, France. Electronic address: david.perrais@u-bordeaux.fr.
authors.forename: David
authors.initials: D
authors.lastname: Perrais
authors.affiliation: Membrane Traffic and Cell Division Lab, Cell Biology and Infection Department, Institut Pasteur, 25-28 Rue du Dr. Roux, 75724 Paris Cedex 15, France; Centre National de la Recherche Scientifique UMR3691, 75015 Paris, France. Electronic address: arnaud.echard@pasteur.fr.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Echard
mesh_terms: D002966:Clathrin / Q000378:metabolism; D022163:Clathrin-Coated Vesicles / Q000378:metabolism; D003034:Coated Pits, Cell-Membrane / Q000378:metabolism; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008840:Microfilament Proteins / Q000378:metabolism; D009800:Oculocerebrorenal Syndrome / Q000201:enzymology / Q000378:metabolism*; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D021381:Protein Transport; D017527:Receptor, IGF Type 2 / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D002966:Clathrin; D008840:Microfilament Proteins; D019269:Phosphatidylinositol 4,5-Diphosphate; D017527:Receptor, IGF Type 2; D010744:Phosphoric Monoester Hydrolases; C075695:OCRL protein, human; C511183:RAB35 protein, human; D020691:rab GTP-Binding Proteins