title: The N-terminal domains determine cellular localization and functions of the Doa4 and Ubp5 deubiquitinating enzymes.
abstract: Ubiquitination is involved in numerous cellular regulatory mechanisms including the cell cycle, signal transduction and quality control. Ubiquitin modifies proteins by consecutive actions of ubiquitin-activating/conjugating enzymes. Attachment of ubiquitin is reversible. Deubiquitinating enzymes are responsible for removal of ubiquitin from ubiquitin-protein conjugates. Genome of the yeast Saccharomyces cerevisiae encodes structurally related but functionally distinct enzymes - Doa4 and Ubp5. Doa4 is involved in general ubiquitin-dependent proteolysis and is responsible for deubiquitination of ubiquitin-protein conjugates at the cytoplasmic face of the late endosome. The N-terminal domain targets the enzyme to the endosome membrane after ESCRT-III complex has formed there. By contrast, corresponding region of homologous Ubp5 is critical for its bud neck localization in dividing cells. Conceivably, Ubp5 plays an essential role in cytokinesis. Here we show that Doa4 physically interacts with the ESCRT-III component Snf7 and preferentially cleaves Lys63-linked ubiquitin oligomers involved in membrane protein trafficking. We also demonstrate that the unstable regulator of cytokinesis Hof1 accumulates in proteasomal mutants and is required for cellular localization of Ubp5.
authors.affiliation: Department of Pharmacology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA.
authors.forename: Nazia
authors.initials: N
authors.lastname: Wolters
authors.affiliation: Department of Pharmacology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA; Emanuel Institute of Biochemical Physics, Russian Academy of Sciences, 4 Kosygin Street, Moscow 119991, Russian Federation. Electronic address: amerik.alexander@gmail.com.
authors.forename: Alexander
authors.initials: A
authors.lastname: Amerik
mesh_terms: D010450:Endopeptidases / Q000737:chemistry / Q000378:metabolism*; D056827:Endosomal Sorting Complexes Required for Transport / Q000737:chemistry / Q000378:metabolism*; D011992:Endosomes / Q000378:metabolism; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism*; D043222:Ubiquitin Thiolesterase / Q000737:chemistry / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: C084434:DOA4 protein, S cerevisiae; D056827:Endosomal Sorting Complexes Required for Transport; D029701:Saccharomyces cerevisiae Proteins; D010450:Endopeptidases; D043222:Ubiquitin Thiolesterase
keywords: Cytokinesis; Deubiquitinating enzymes; ESCRT-III; Ubiquitin; Vacuolar protein sorting