Cytoplasmic dynein participates in the centrosomal localization of the Golgi complex.
Cytoplasmic dynein participates in the centrosomal localization of the Golgi complex.
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DOI：
10.1083/jcb.118.6.1333
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发表时间：
1992-09
期刊：
影响因子：
--
通讯作者：
Pfeffer SR
中科院分区：
文献类型：
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作者：
Corthésy-Theulaz I;Pauloin A;Pfeffer SR
高尔基复合体的定位取决于微管装置的完整性。在间期，高尔基体的中心粒周围定位受到限制。在有丝分裂过程中，它分裂成小囊泡，分散在整个细胞质中，直到末期，此时它们再次在中心体附近聚结。这些观察结果表明，高尔基复合体利用类似动力蛋白的马达来介导其从细胞外周向位于中心体的微管负端的运输。我们利用半完整的细胞来研究高尔基复合体与微管装置的相互作用。我们在这里表明高尔基复合体可以进入半完整的细胞并与细胞质成分稳定结合。稳定的结合在这里被称为“高尔基体捕获”，需要 ATP 水解和完整的微管，并且在添加胞质蛋白的情况下在生理温度下最大程度地发生。一旦转移到半完整细胞的细胞质中，外源高尔基复合体在微管组织中心附近表现出与内源高尔基复合体相似的分布。高尔基体捕获过程需要细胞质微管蛋白，如果细胞质动力蛋白从细胞质中被免疫耗尽，则高尔基体捕获过程就会被取消。由 CHO 细胞胞质溶胶制备的细胞质动力蛋白可恢复用动力蛋白免疫耗竭的胞质溶胶进行的反应的高尔基体捕获活性。这些结果表明细胞质动力蛋白可以与分离的高尔基复合体相互作用，并参与它们在半完整受体细胞中心体附近的积累。因此，细胞质动力蛋白似乎在确定高尔基复合体的亚细胞定位中发挥作用。
The localization of the Golgi complex depends upon the integrity of the microtubule apparatus. At interphase, the Golgi has a restricted pericentriolar localization. During mitosis, it fragments into small vesicles that are dispersed throughout the cytoplasm until telophase, when they again coalesce near the centrosome. These observations have suggested that the Golgi complex utilizes a dynein-like motor to mediate its transport from the cell periphery towards the minus ends of microtubules, located at the centrosome. We utilized semi-intact cells to study the interaction of the Golgi complex with the microtubule apparatus. We show here that Golgi complexes can enter semi-intact cells and associate stably with cytoplasmic constituents. Stable association, termed here "Golgi capture," requires ATP hydrolysis and intact microtubules, and occurs maximally at physiological temperature in the presence of added cytosolic proteins. Once translocated into the semi-intact cell cytoplasm, exogenous Golgi complexes display a distribution similar to endogenous Golgi complexes, near the microtubule-organizing center. The process of Golgi capture requires cytoplasmic tubulin, and is abolished if cytoplasmic dynein is immunodepleted from the cytosol. Cytoplasmic dynein, prepared from CHO cell cytosol, restores Golgi capture activity to reactions carried out with dynein immuno-depleted cytosol. These results indicate that cytoplasmic dynein can interact with isolated Golgi complexes, and participate in their accumulation near the centrosomes of semi-intact, recipient cells. Thus, cytoplasmic dynein appears to play a role in determining the subcellular localization of the Golgi complex.