The Vps13p-Cdc31p complex is directly required for TGN late endosome transport and TGN homotypic fusion.
The Vps13p-Cdc31p complex is directly required for TGN late endosome transport and TGN homotypic fusion.
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DOI：
10.1083/jcb.201606078
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发表时间：
2017-02
期刊：
影响因子：
--
通讯作者：
Fuller RS
中科院分区：
文献类型：
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作者：
De M;Oleskie AN;Ayyash M;Dutta S;Mancour L;Abazeed ME;Brace EJ;Skiniotis G;Fuller RS
VPS 13蛋白在真核生物中广泛保守，并与人类神经退行性疾病和神经发育疾病相关。De等人描述了酵母Vps 13 p的脂质特异性和结构，提供了对其在TGN晚期内体转运和TGN同型融合中的作用的深入了解。酵母VPS 13是在细胞生物学和医学中越来越感兴趣的真核基因家族的创始成员。四个人类VPS 13基因中的三个突变导致常染色体隐性神经退行性或神经发育疾病，使酵母VPS 13 p成为重要的结构和功能模型。使用纯化的Vps 13 p的无细胞重建，我们表明，Vps 13 p是直接需要从trans-Golgi网络（TGN）运输到晚期内体/prevacuolar室（PVC）和TGN同型融合。Vps 13 p必须与小的钙结合蛋白Cdc 31 p复合才有活性。负染的Vps 13 p的单粒子电子显微镜分析表明，该358-kD蛋白折叠成紧凑的棒状密度（20 × 4 nm），其一端具有环状结构，环状开口直径约为6 nm。Vps 13 p表现出ATP刺激的结合酵母细胞膜和特定的相互作用与磷脂酸和磷酸化形式的磷脂酰肌醇至少部分通过保守的N-和C-末端结构域的结合亲和力。
VPS13 proteins are widely conserved in eukaryotes and associated with human neurodegenerative and neurodevelopmental diseases. De et al. describe the lipid specificity and structure of yeast Vps13p, providing insight into its role in both TGN late endosome transport and TGN homotypic fusion. 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.