title: The Vps13p-Cdc31p complex is directly required for TGN late endosome transport and TGN homotypic fusion.
abstract: Yeast VPS13 is the founding member of a eukaryotic gene family of growing interest in cell biology and medicine. Mutations in three of four human VPS13 genes cause autosomal recessive neurodegenerative or neurodevelopmental disease, making yeast Vps13p an important structural and functional model. Using cell-free reconstitution with purified Vps13p, we show that Vps13p is directly required both for transport from the trans-Golgi network (TGN) to the late endosome/prevacuolar compartment (PVC) and for TGN homotypic fusion. Vps13p must be in complex with the small calcium-binding protein Cdc31p to be active. Single-particle electron microscopic analysis of negatively stained Vps13p indicates that this 358-kD protein is folded into a compact rod-shaped density (20 × 4 nm) with a loop structure at one end with a circular opening ∼6 nm in diameter. Vps13p exhibits ATP-stimulated binding to yeast membranes and specific interactions with phosphatidic acid and phosphorylated forms of phosphatidyl inositol at least in part through the binding affinities of conserved N- and C-terminal domains.
authors.affiliation: Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109.
authors.forename: Mithu
authors.initials: M
authors.lastname: De
authors.affiliation: Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109.|Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Austin N
authors.initials: AN
authors.lastname: Oleskie
authors.affiliation: Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109.
authors.forename: Mariam
authors.initials: M
authors.lastname: Ayyash
authors.affiliation: Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109.|Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Somnath
authors.initials: S
authors.lastname: Dutta
authors.affiliation: Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109.|Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Liliya
authors.initials: L
authors.lastname: Mancour
authors.affiliation: Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109.|Medical Scientist Training Program, University of Michigan Medical School, Ann Arbor, MI 48109.
authors.forename: Mohamed E
authors.identifier: 0000-0002-6614-4440
authors.initials: ME
authors.lastname: Abazeed
authors.affiliation: Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109.
authors.forename: Eddy J
authors.initials: EJ
authors.lastname: Brace
authors.affiliation: Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109.|Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Georgios
authors.initials: G
authors.lastname: Skiniotis
authors.affiliation: Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109 bfuller@umich.edu.
authors.forename: Robert S
authors.identifier: 0000-0003-2584-8246
authors.initials: RS
authors.lastname: Fuller
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011992:Endosomes / Q000378:metabolism*; D005838:Genotype; D006056:Golgi Apparatus / Q000378:metabolism*; D007425:Intracellular Membranes / Q000378:metabolism*; D008561:Membrane Fusion; D046912:Multiprotein Complexes; D009154:Mutation; D010641:Phenotype; D010712:Phosphatidic Acids / Q000378:metabolism; D010716:Phosphatidylinositols / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D017510:Protein Folding; D054730:Protein Interaction Domains and Motifs; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013329:Structure-Activity Relationship; D013997:Time Factors
chemical_list: C083769:CDC31 protein, S cerevisiae; D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D046912:Multiprotein Complexes; D010712:Phosphatidic Acids; D010716:Phosphatidylinositols; D029701:Saccharomyces cerevisiae Proteins; C110276:VPS13 protein, S cerevisiae; D000255:Adenosine Triphosphate