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Journal of Cell Science, Vol 110, Issue 19 2495-2505, Copyright © 1997 by Company of Biologists
JOURNAL ARTICLES |
AA Minin
Institute of Protein Research, Russian Academy of Sciences, Moscow. minin@imb.imb.ac.ru
The morphology and location of the Golgi apparatus (GA) has been shown to change upon microtubule (Mt) depolymerization. The GA in different cell types undergoes fragmentation and dispersal throughout the cytoplasm upon treatment with nocodazole. In this study experiments were performed on human skin fibroblasts (HSFs) and rat fibroblasts (REF 52) to determine whether the dispersal of GA in HSFs treated with nocodazole is dependent on Mts that show the higher resistance to this Mt-depolymerizing drug. It is shown here that nocodazole at concentrations as low as 100 nM caused the GA to disperse in treated fibro-blasts that still contained a fairly high amount of Mts. Antibody-blocking analysis of Mts after injection of biotin-tubulin into the HSFs was used to show that nocodazole at low concentrations induced the stabilization of the remaining Mts. The complete disruption of Mts by the incubation of HSFs at 0 degrees C prevented the dispersal of GA from the perinuclear area when the cells were subsequently warmed to 37 degrees C in the presence of nocodazole. Micro-injection of the well-characterized HD antibody against kinesin but not the preimmune IgG caused inhibition of GA dispersal in HSFs by nocodazole. These data demonstrate that the dispersal of GA in the cytoplasm of nocodazole-treated HSFs is a kinesin-driven process with stable Mts serving as tracks.
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