A role for Rab10 in von Willebrand factor release discovered by an AP-1 interactor screen in C-elegans
A role for Rab10 in von Willebrand factor release discovered by an AP-1 interactor screen in C-elegans
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DOI：
10.1111/j.1538-7836.2010.04138.x
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发表时间：
2011-02-01
影响因子：
10.4
通讯作者：
Cutler, D. F.
中科院分区：
文献类型：
--
作者：
Michaux, G.;Dyer, C. E. F.;Cutler, D. F.
背景：内皮血管性血友病因子（VWF）介导血小板粘附，并作为凝血因子VIII的保护性伴侣。高度多聚化的VWF的快速释放在促进止血方面特别有效。为了产生这种蛋白质，需要一个精心设计的生物发生，最终在称为韦伯-帕拉德体（WPB）的分泌颗粒内储存的高尔基体网络（TGN）中达到顶峰。不能正确形成这些细胞器可导致VWF的低分子量多聚体的不受控制的分泌。与TGN相关的接头AP-1及其相互作用物网格蛋白、aftiphilin和γ-协同素对于高尔基体初始WPB形成以及VWF储存和分泌是必不可少的。目的：鉴定与VWF储存和/或分泌有关的新蛋白。方法：利用全基因组RNA干扰（RNAi）技术筛选线虫C。elegans来鉴定新的AP-1基因相互作用物。结果：发现小GT-Rab 10与C.优雅的。我们研究了Rab 10在人原代脐静脉内皮细胞（HUVECs）中的作用。我们报告说，Rab 10是丰富的高尔基体，WPB的形成，并在Rab 10的表达已被siRNA抑制的细胞中，VWF分泌被改变：快速释放的VWF的量显着减少。我们还发现Rab 8A具有类似的功能。结论：Rab 10和Rab 8A是参与WPB生物发生的新的细胞质因子，在产生可快速响应促分泌素的颗粒中发挥作用。
Background: Endothelial von Willebrand factor (VWF) mediates platelet adhesion and acts as a protective chaperone to clotting factor VIII. Rapid release of highly multimerized VWF is particularly effective in promoting hemostasis. To produce this protein, an elaborate biogenesis is required, culminating at the trans-Golgi network (TGN) in storage within secretory granules called Weibel-Palade bodies (WPB). Failure to correctly form these organelles can lead to uncontrolled secretion of low-molecular-weight multimers of VWF. The TGN-associated adaptor AP-1 and its interactors clathrin, aftiphilin and gamma-synergin are essential to initial WPB formation at the Golgi apparatus, and thus to VWF storage and secretion. Objectives: To identify new proteins implicated in VWF storage and/or secretion. Methods: A genomewide RNA interference (RNAi) screen was performed in the Nematode C. elegans to identify new AP-1 genetic interactors. Results: The small GTPase Rab10 was found to genetically interact with a partial loss of function of AP-1 in C. elegans. We investigated Rab10 in human primary umbilical vein endothelial cells (HUVECs). We report that Rab10 is enriched at the Golgi apparatus, where WPB are formed, and that in cells where Rab10 expression has been suppressed by siRNA, VWF secretion is altered: the amount of rapidly released VWF was significantly reduced. We also found that Rab8A has a similar function. Conclusion: Rab10 and Rab8A are new cytoplasmic factors implicated in WPB biogenesis that play a role in generating granules that can rapidly respond to secretagogue.