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The Golgi apparatus of higher eukaryotes is a dynamic organelle. Recent studies have demonstrated that kinesin-dependent transport of vesicular cargo is a key mechanism for maintaining normal Golgi morphology and function. However, how cargo is transported to the Golgi region remains unknown. Using the COS7 and L cell lines as model systems, we examined whether kinesin-based intracellular transport may be involved in the initiation of Golgi dispersal in the early secretory pathway. A method was developed to synchronously disperse Golgi apparatus within cultured cells. This method involves the activation of the p38 mitogen-activated protein kinase (MAPK) pathway and the initiation of the cell cycle by stimulation with ultraviolet B radiation and serum. We demonstrate that, in cells transiently expressing GFP-fused kinesin light-chain protein or cargo (galactosyltransferase II), Golgi dispersal was observed only when cells were arrested in the late G(1)/early S phase of the cell cycle. Golgi dispersal was also observed in cells expressing two-hybrid-dependent beta-galactosidase, a fusion of p38 MAPK and beta-galactosidase with a mutant of the catalytic domain of the m-calpain. Our results suggest that cargo transport is required for Golgi dispersal, but the molecular details of this transport remain unknown.One mile in your life
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