Chabin-Brion K, Marceiller J, Perez F, Settegrana C, Drechou A, Durand G, Poüs C
Laboratoire de Biochimie et de Biologie Cellulaire, EA 1595, Faculté de Pharmacie, 92296 Châtenay-Malabry Cedex, France.
Mol Biol Cell. 2001 Jul;12(7):2047-60. doi: 10.1091/mbc.12.7.2047.
We show that the Golgi complex can directly stimulate microtubule nucleation in vivo and in vitro and thus behaves as a potent microtubule-organizing organelle in interphase cells. With the use of nocodazole wash-out experiments in hepatic cells, we found that the occurrence of noncentrosomal, early stabilized microtubules is highly correlated with the subcellular localization of Golgi membranes. With the use of in vitro reconstituted microtubule assembly systems with or without cytosol, we also found that, in contrast to centrosomally attached microtubules, the distal ends of Golgi-attached microtubules are remotely stabilized in a way that requires additional cytosolic component(s). Finally, we demonstrate that Golgi-based microtubule nucleation is direct and involves a subset of gamma-tubulin bound to the cytoplasmic face of the organelle.
我们发现,高尔基体复合物在体内和体外均可直接刺激微管成核,因此在间期细胞中表现为一种强大的微管组织细胞器。通过在肝细胞中进行诺考达唑洗脱实验,我们发现非中心体的早期稳定微管的出现与高尔基体膜的亚细胞定位高度相关。通过使用含有或不含有胞质溶胶的体外重组微管组装系统,我们还发现,与中心体附着的微管不同,高尔基体附着的微管的远端以一种需要额外胞质成分的方式远距离稳定。最后,我们证明基于高尔基体的微管成核是直接的,并且涉及与该细胞器胞质面结合的γ-微管蛋白的一个子集。