Recruitment of CG-NAP to the Golgi apparatus through interaction with dynein-dynactin complex
Recruitment of CG-NAP to the Golgi apparatus through interaction with dynein-dynactin complex
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DOI：
10.1111/j.1365-2443.2007.01055.x
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发表时间：
2007-03-01
期刊：
影响因子：
2.1
通讯作者：
Ono, Yoshitaka
中科院分区：
文献类型：
--
作者：
Kim, Hon-Song;Takahashi, Mikiko;Ono, Yoshitaka
高尔基体的结构组织和位置受到微管细胞骨架和微管马达蛋白的高度调节。这些蛋白质与高尔基体的连接机制仍然是难以捉摸的。在这里，我们发现，中心体和高尔基体定位PKN相关蛋白（CG-NAP）被定位到高尔基体中的微管依赖性的方式。微管结合实验表明，CG-NAP具有两个微管结合结构域。我们还发现，CG-NAP是很好的共定位与细胞质动力蛋白亚基在恢复过程中从冰上处理的细胞，诱导解离的CG-NAP从高尔基体。在从乙酸盐处理恢复期间观察到类似的共定位，已报道乙酸盐处理抑制动力蛋白介导的转运。CG-NAP与动力蛋白亚基p150（Glued）共免疫沉淀。表达CG-NAP的p150（Glued）结合区融合的靶向序列诱导的线粒体的招聘到pericentriolar区，这表明该区域与功能性细胞质动力蛋白在体内相互作用。此外，该区域的过度表达导致类似于dynamitin的高尔基体碎片化。这些结果表明，CG-NAP被招募到微管的负端与细胞质动力蛋白的相互作用，从而定位到高尔基体的微管依赖性的方式，并可能参与高尔基体附近的中心体的形成。
The structural organization and position of the Golgi apparatus are highly regulated by microtubule cytoskeleton and microtubule motor proteins. The mechanisms linking these proteins to the Golgi apparatus remain elusive. Here, we found that centrosome and Golgi-localized PKN associated protein (CG-NAP) was localized to the Golgi apparatus in a microtubule-dependent manner. Microtubule-binding experiments revealed that CG-NAP possessed two microtubule-binding domains. We also found that CG-NAP was well co-localized with cytoplasmic dynein subunits during recovery from the on-ice treatment of cells that induced dissociation of CG-NAP from the Golgi. Similar co-localization was observed during recovery from the acetate treatment, which has been reported to inhibit the dynein-mediated transport. CG-NAP was co-immunoprecipitated with a dynactin subunit p150(Glued). Expressing the p150(Glued)-binding region of CG-NAP fused with mitochondria-targeting sequence induced recruitment of mitochondria to the pericentriolar area, suggesting that this region interacts with functional cytoplasmic dynein in vivo. Moreover, over-expression of this region caused fragmentation of the Golgi similar to that of dynamitin. These results suggest that CG-NAP is recruited to the minus ends of microtubules by interacting with cytoplasmic dynein, thereby localizes to the Golgi apparatus in a microtubule-dependent manner and possibly involved in the formation of the Golgi near the centrosomes.