title: JosD1, a membrane-targeted deubiquitinating enzyme, is activated by ubiquitination and regulates membrane dynamics, cell motility, and endocytosis.
abstract: The functional diversity of deubiquitinating enzymes (DUBs) is not well understood. The MJD family of DUBs consists of four cysteine proteases that share a catalytic "Josephin" domain. The family is named after the DUB ATXN3, which causes the neurodegenerative disease Machado-Joseph disease. The two closely related Josephin domain-containing (JosD) proteins 1 and 2 consist of little more than the Josephin domain. To gain insight into the properties of Josephin domains, we investigated JosD1 and JosD2. JosD1 and JosD2 were found to differ fundamentally in many respects. In vitro, only JosD2 can cleave ubiquitin chains. In contrast, JosD1 cleaves ubiquitin chains only after it is monoubiquitinated, a form of posttranslational-dependent regulation shared with ATXN3. A significant fraction of JosD1 is monoubiquitinated in diverse mouse tissues. In cell-based studies, JosD2 localizes to the cytoplasm whereas JosD1 preferentially localizes to the plasma membrane, particularly when ubiquitinated. The membrane occupancy by JosD1 suggests that it could participate in membrane-dependent events such as cell motility and endocytosis. Indeed, time-lapse imaging revealed that JosD1 enhances membrane dynamics and cell motility. JosD1 also influences endocytosis in cultured cells by increasing the uptake of endocytic markers of macropinocytosis while decreasing those for clathrin- and caveolae-mediated endocytosis. Our results establish that two closely related DUBs differ markedly in activity and function and that JosD1, a membrane-associated DUB whose activity is regulated by ubiquitination, helps regulate membrane dynamics, cell motility, and endocytosis.
authors.affiliation: Department of Neurology, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Seki
authors.forename: Lijie
authors.initials: L
authors.lastname: Gong
authors.forename: Aislinn J
authors.initials: AJ
authors.lastname: Williams
authors.forename: Norio
authors.initials: N
authors.lastname: Sakai
authors.forename: Sokol V
authors.initials: SV
authors.lastname: Todi
authors.forename: Henry L
authors.initials: HL
authors.lastname: Paulson
mesh_terms: D000818:Animals; D019556:COS Cells; D021941:Caveolae / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology*; D002465:Cell Movement; D002522:Chlorocebus aethiops; D002966:Clathrin / Q000378:metabolism; D004705:Endocytosis; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D009928:Organ Specificity; D021381:Protein Transport; D059010:Single-Cell Analysis; D059008:Time-Lapse Imaging; D054875:Ubiquitination
chemical_list: D002966:Clathrin; D010450:Endopeptidases; C582950:JosD1 protein, mouse
keywords: Ataxin-3; Cell Motility; Deubiquitinating Enzyme; Deubiquitination; Endocytosis; Josephin Domain-containing Protein 1; Membrane Dynamics; Ubiquitin; Ubiquitination