Arabidopsis dynamin-related proteins, DRP2A and DRP2B, function coordinately in post-Golgi trafficking
Arabidopsis dynamin-related proteins, DRP2A and DRP2B, function coordinately in post-Golgi trafficking
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DOI：
10.1016/j.bbrc.2014.11.065
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发表时间：
2015-01-02
影响因子：
3.1
通讯作者：
Tsutsumi, Nobuhiro
中科院分区：
文献类型：
--
作者：
Huang, Jiahe;Fujimoto, Masaru;Tsutsumi, Nobuhiro
动力蛋白相关蛋白(Dynamin-Related Proteins，DRPs)是一类大的GTP酶，参与广泛的细胞膜重构过程。在拟南芥中，两个近缘陆地植物特有的DRP2A和DRP2B被认为参与了高尔基体后运输的调控。在这里，我们研究了它们的分子性质和功能关系。QRT-PCR和GUS分析表明，DRP2A和DRP2B在根尖分生组织和维管束附近表达最强，但表达普遍。酵母双杂交、双分子荧光互补和免疫共沉淀质谱分析表明，DRP2A和DRP2B相互作用。激光共聚焦扫描显微镜和可变入射角荧光显微镜观察到，DRP2A和DRP2B的荧光融合几乎完全共定位，并且主要定位于根表皮细胞质膜、富含网状蛋白的反高尔基体网和细胞板的内吞囊泡形成部位。用磷脂酰肌醇3-/4-激酶抑制剂Wortmannin、肌动蛋白聚合抑制剂Latrunculin B和微管聚合抑制剂oryzalin处理后，DRP2A和DRP2B在质膜上的停留时间增加。这些结果表明，DRP2A和DRP2B在高尔基体后转运磷脂酰肌醇3-或4-激酶和细胞骨架聚合依赖的多条途径中起协调作用。(C)2014 Elsevier Inc.保留所有权利。
Dynamin-related proteins (DRPs) are large GTPases involved in a wide range of cellular membrane remodeling processes. In Arabidopsis thaliana, two paralogous land plant-specific type DRPs, DRP2A and DRP2B, are thought to participate in the regulation of post-Golgi trafficking. Here, we examined their molecular properties and functional relationships. qRT-PCR and GUS assays showed that DRP2A and DRP2B were expressed ubiquitously, although their expressions were strongest around root apical meristems and vascular bundles. Yeast two-hybrid, bi-molecular fluorescent complementation, and co-immunoprecipitation mass spectrometry analyses revealed that DRP2A and DRP2B interacted with each other. In observations with confocal laser scanning microscopy and variable incidence angle fluorescent microscopy, fluorescent fusions of DRP2A and DRP2B almost completely co-localized and were mainly localized to endocytic vesicle formation sites of the plasma membrane, clathrin-enriched trans-Golgi network and the cell plate in root epidermal cells. Treatments with wortmannin, an inhibitor of phosphatidylinositol 3-/4-kinases, latrunculin B, an inhibitor of actin polymerization, and oryzalin, an inhibitor of microtubule polymerization, increased the resident time of DRP2A and DRP2B on the plasma membrane. These results show that DRP2A and DRP2B function coordinately in multiple pathways of post-Golgi trafficking in phosphatidylinositol 3- or 4-kinase and cytoskeleton polymerization-dependent manners. (C) 2014 Elsevier Inc. All rights reserved.