The Vps13p-Cdc31p complex is directly required for TGN late endosome transport and TGN homotypic fusion

• Published in: The Journal of cell biology Vol. 216; no. 2; pp. 425 - 439
• Main Authors: De, Mithu, Oleskie, Austin N, Ayyash, Mariam, Dutta, Somnath, Mancour, Liliya, Abazeed, Mohamed E, Brace, Eddy J, Skiniotis, Georgios, Fuller, Robert S
• Format: Journal Article
• Language: English
• Published: United States Rockefeller University Press 01.02.2017; The Rockefeller University Press
• Subjects: Adenosine Triphosphate - metabolism; Calcium-Binding Proteins - chemistry; Calcium-Binding Proteins - genetics; Calcium-Binding Proteins - metabolism; Cell Cycle Proteins - chemistry; Cell Cycle Proteins - genetics; Cell Cycle Proteins - metabolism; Cellular biology; Endosomes - metabolism; Eukaryotes; Genes; Genotype; Golgi Apparatus - metabolism; Intracellular Membranes - metabolism; Membrane Fusion; Membranes; Multiprotein Complexes; Mutation; Phenotype; Phosphatidic Acids - metabolism; Phosphatidylinositols - metabolism; Phosphorylation; Protein Binding; Protein Folding; Protein Interaction Domains and Motifs; Protein Transport; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - chemistry; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; Signal Transduction; Structure-Activity Relationship; Time Factors; Yeast
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.201606078

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.