Stacks on tracks: the plant Golgi apparatus traffics on an actin/ER network
Stacks on tracks: the plant Golgi apparatus traffics on an actin/ER network
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DOI：
10.1046/j.1365-313x.1998.00208.x
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发表时间：
1998-08-01
期刊：
影响因子：
7.2
通讯作者：
Hawes, C
中科院分区：
文献类型：
--
作者：
Boevink, P;Oparka, K;Hawes, C
我们通过两种高尔基蛋白与绿色荧光蛋白（GFP）的融合表达，可视化了内质网（ER）与克利夫兰烟草（Nicotiana clevelandii）叶细胞高尔基体的关系。将大鼠唾液转移酶的跨膜结构域（信号锚定序列）与GFP融合到高尔基体上。第二个表达拟南芥H/KDEL受体同源物aERD2的构建体与GFP融合，同时靶向高尔基体和内质网，从而首次在活细胞中研究这两种细胞器之间的关系。高尔基堆积被证明沿着叶表皮细胞的多边形皮质内质网快速而广泛地移动，而不离开内质网小管。在表达gfp的细胞中，f -肌动蛋白的go定位揭示了一个与内质网结构精确匹配的潜在肌动蛋白细胞骨架，而用细胞松弛素D和n -乙基马来酰亚胺抑制剂处理的细胞揭示了高尔基运动对肌动蛋白索的依赖性。这些观察结果表明，叶片高尔基复合体的功能是作为一个肌动蛋白导向堆栈的运动系统，其功能是从相对静止的内质网系统中拾取产物。此外，我们首次在体内证明了brefeldin a诱导高尔基膜蛋白向内质网的逆行转运。
We have visualized the relationship between the endoplasmic reticulum (ER) and Golgi in leaf cells of Nicotiana clevelandii by expression of two Golgi proteins fused to green fluorescent protein (GFP). A fusion of the transmembrane domain (signal anchor sequence) of a rat sialyl transferase to GFP was targeted to the Golgi stacks. A second construct that expressed the Arabidopsis H/KDEL receptor homologue aERD2, fused to GFP, was targeted to both the Golgi apparatus and ER, allowing the relationship between these two organelles to be studied in living cells for the first time. The Golgi stacks were shown to move rapidly and extensively along the polygonal cortical ER network of leaf epidermal cells, without departing from the ER tubules. Go-localization of F-actin in the GFP-expressing cells revealed an underlying actin cytoskeleton that matched precisely the architecture of the ER network, while treatment of cells with the inhibitors cytochalasin D and N-ethylmaleimide revealed the dependency of Golgi movement on actin cables. These observations suggest that the leaf Golgi complex functions as a motile system of actin-directed stacks whose function is to pick up products from a relatively stationary ER system. Also, we demonstrate for the first time in vivo brefeldin A-induced retrograde transport of Golgi membrane protein to the ER.