title: Dynamin 2 mutations associated with human diseases impair clathrin-mediated receptor endocytosis.
abstract: Dynamin 2 (DNM2) is a large GTPase involved in the release of nascent vesicles during endocytosis and intracellular membrane trafficking. Distinct DNM2 mutations, affecting the middle domain (MD) and the Pleckstrin homology domain (PH), have been identified in autosomal dominant centronuclear myopathy (CNM) and in the intermediate and axonal forms of the Charcot-Marie-Tooth peripheral neuropathy (CMT). We report here the first CNM mutation (c.1948G>A, p.E650 K) in the DNM2 GTPase effector domain (GED), leading to a slowly progressive moderate myopathy. COS7 cells transfected with DNM2 constructs harboring a disease-associated mutation in MD, PH, or GED show a reduced uptake of transferrin and low-density lipoprotein (LDL) complex, two markers of clathrin-mediated receptor endocytosis. A decrease in clathrin-mediated endocytosis was also identified in skin fibroblasts from one CNM patient. We studied the impact of DNM2 mutant overexpression on epidermal growth factor (EGF)-induced extracellular signal-regulated kinase 1 (ERK1) and ERK2 activation, known to be an endocytosis- and DNM2-dependent process. Activation of ERK1/2 was impaired for all the transfected mutants in COS7 cells, but not in CNM fibroblasts. Our results indicate that impairment of clathrin-mediated endocytosis may play a role in the pathophysiological mechanisms leading to DNM2-related diseases, but the tissue-specific impact of DNM2 mutations in both diseases remains unclear.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, U582, Institut de Myologie, Paris, France. m.bitoun@institut-myologie.org
authors.forename: Marc
authors.initials: M
authors.lastname: Bitoun
authors.forename: Anne-Cécile
authors.initials: AC
authors.lastname: Durieux
authors.forename: Bernard
authors.initials: B
authors.lastname: Prudhon
authors.forename: Jorge A
authors.initials: JA
authors.lastname: Bevilacqua
authors.forename: Adrien
authors.initials: A
authors.lastname: Herledan
authors.forename: Vehary
authors.initials: V
authors.lastname: Sakanyan
authors.forename: Andoni
authors.initials: A
authors.lastname: Urtizberea
authors.forename: Luis
authors.initials: L
authors.lastname: Cartier
authors.forename: Norma B
authors.initials: NB
authors.lastname: Romero
authors.forename: Pascale
authors.initials: P
authors.lastname: Guicheney
mesh_terms: D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D019556:COS Cells; D002522:Chlorocebus aethiops; D002966:Clathrin / Q000502:physiology*; D017931:DNA Primers; D034285:Dynamin II / Q000235:genetics*; D004705:Endocytosis / Q000235:genetics*; D004789:Enzyme Activation; D004815:Epidermal Growth Factor / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D009154:Mutation; D014168:Transferrin / Q000378:metabolism
chemical_list: D002966:Clathrin; D017931:DNA Primers; D014168:Transferrin; D004815:Epidermal Growth Factor; D048049:Extracellular Signal-Regulated MAP Kinases; D034285:Dynamin II