Actin dynamics coupled to clathrin-coated vesicle formation at the trans-Golgi network.
Actin dynamics coupled to clathrin-coated vesicle formation at the trans-Golgi network.
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DOI：
10.1083/jcb.200403120
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发表时间：
2004-06-21
期刊：
影响因子：
--
通讯作者：
Drubin DG
中科院分区：
文献类型：
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作者：
Carreno S;Engqvist-Goldstein AE;Zhang CX;McDonald KL;Drubin DG
在不同的物种中，肌动蛋白的组装促进了胞吞作用中笼状蛋白涂层小泡(CCV)的形成。这一作用可能是对细胞皮质独特环境的一种适应，也可能是促进CCV在不同膜上形成的基础。Sla2p/Hip1R家族的蛋白在酵母和哺乳动物的胞内位置与肌动蛋白和网状蛋白结合。我们推测，Hip1R还可能协调肌动蛋白组装和跨高尔基体网络(TGN)上的网状蛋白出芽。使用去卷积和时间推移显微镜，我们发现Hip1R存在于从TGN产生的CCV上。这些囊泡含有参与将蛋白质靶向溶酶体的甘露糖6-磷酸受体，以及肌动蛋白核化Arp2/3复合体。通过RNAi沉默Hip1R的表达导致高尔基体组织的破坏和与TGN上CCV相关的F-肌动蛋白结构的积累。Hip1R沉默和肌动蛋白毒物使组织蛋白D退出TGN的速度减慢。这些研究确定了Hip1R和肌动蛋白在CCV从TGN萌发的溶酶体生物发生中的作用。
In diverse species, actin assembly facilitates clathrin-coated vesicle (CCV) formation during endocytosis. This role might be an adaptation specific to the unique environment at the cell cortex, or it might be fundamental, facilitating CCV formation on different membranes. Proteins of the Sla2p/Hip1R family bind to actin and clathrin at endocytic sites in yeast and mammals. We hypothesized that Hip1R might also coordinate actin assembly with clathrin budding at the trans-Golgi network (TGN). Using deconvolution and time-lapse microscopy, we showed that Hip1R is present on CCVs emerging from the TGN. These vesicles contain the mannose 6-phosphate receptor involved in targeting proteins to the lysosome, and the actin nucleating Arp2/3 complex. Silencing of Hip1R expression by RNAi resulted in disruption of Golgi organization and accumulation of F-actin structures associated with CCVs on the TGN. Hip1R silencing and actin poisons slowed cathepsin D exit from the TGN. These studies establish roles for Hip1R and actin in CCV budding from the TGN for lysosome biogenesis.