Kinesin is the motor for microtubule-mediated Golgi-to-ER membrane traffic.
Kinesin is the motor for microtubule-mediated Golgi-to-ER membrane traffic.
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DOI：
10.1083/jcb.128.3.293
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发表时间：
1995-02
期刊：
影响因子：
--
通讯作者：
Bloom GS
中科院分区：
文献类型：
--
作者：
Lippincott-Schwartz J;Cole NB;Marotta A;Conrad PA;Bloom GS
动物细胞中内质网和高尔基复合体的分布和动力学都依赖于微管;在许多细胞类型中，内质网向细胞周边微管的正端延伸，高尔基簇在中心体附近微管的负端。在这项研究中，我们提供的证据表明，微管电机，驱动蛋白，是目前的ER和高尔基体之间的膜循环和权力在这个系统内的膜外周定向运动。在NRK细胞中使用驱动蛋白重链的H1单克隆抗体在光镜和电镜水平上对驱动蛋白进行免疫定位，揭示驱动蛋白与ER/高尔基体系统的所有膜相关。然而，在37摄氏度的稳态下，驱动蛋白最集中在外周分布的前高尔基体结构上，所述前高尔基体结构含有从ER新释放的β COP和水疱性口炎病毒糖蛋白。在温度降低或诺考达唑处理后，驱动蛋白的分布转移到高尔基体上，而布雷菲德菌素A（BFA）处理，驱动蛋白可以在高尔基体衍生的小管和ER中找到。这表明驱动蛋白与ER和高尔基体之间组成性循环的膜相关。用微量注射驱动蛋白抗体的方法检测驱动蛋白在细胞膜上的作用。高尔基体到内质网而不是内质网到高尔基体的膜转运被发现被显微注射的抗驱动蛋白所抑制，这表明驱动蛋白为微管加上膜到内质网的末端定向再循环提供动力，并且对向高尔基体复合体移动的前高尔基体中间体保持无活性。
The distribution and dynamics of both the ER and Golgi complex in animal cells are known to be dependent on microtubules; in many cell types the ER extends toward the plus ends of microtubules at the cell periphery and the Golgi clusters at the minus ends of microtubules near the centrosome. In this study we provide evidence that the microtubule motor, kinesin, is present on membranes cycling between the ER and Golgi and powers peripherally directed movements of membrane within this system. Immunolocalization of kinesin at both the light and electron microscopy levels in NRK cells using the H1 monoclonal antibody to kinesin heavy chain, revealed kinesin to be associated with all membranes of the ER/Golgi system. At steady-state at 37 degrees C, however, kinesin was most concentrated on peripherally distributed, pre- Golgi structures containing beta COP and vesicular stomatitis virus glycoprotein newly released from the ER. Upon temperature reduction or nocodazole treatment, kinesin's distribution shifted onto the Golgi, while with brefeldin A (BFA)-treatment, kinesin could be found in both Golgi-derived tubules and in the ER. This suggested that kinesin associates with membranes that constitutively cycle between the ER and Golgi. Kinesin's role on these membranes was examined by microinjecting kinesin antibody. Golgi-to-ER but not ER-to-Golgi membrane transport was found to be inhibited by the microinjected anti-kinesin, suggesting kinesin powers the microtubule plus end-directed recycling of membrane to the ER, and remains inactive on pre-Golgi intermediates that move toward the Golgi complex.