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J. Cell Biol.
© The Rockefeller University Press
0021-9525/97/12/1137/19 $2.00
Volume 139, Number 5, December 1, 1997 1137-1155

Golgi Tubule Traffic and the Effects of Brefeldin A Visualized in Living Cells

Noah Sciaky,* John Presley,* Carolyn Smith,Dagger Kristien J.M. Zaal,* Nelson Cole,* Jorge E. Moreira,* Mark Terasaki,Dagger § Eric Siggia,par and Jennifer Lippincott-Schwartz*

* Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892; Dagger  National Institute of Neurological Disorders and Stroke, NIH, Bethesda, Maryland 20892; § Department of Physiology, University of Connecticut Health Center, Farmington, Connecticut 06032; and par  Department of Physics, Cornell University, Ithaca, New York 14853

The Golgi complex is a dynamic organelle engaged in both secretory and retrograde membrane traffic. Here, we use green fluorescent protein-Golgi protein chimeras to study Golgi morphology in vivo. In untreated cells, membrane tubules were a ubiquitous, prominent feature of the Golgi complex, serving both to interconnect adjacent Golgi elements and to carry membrane outward along microtubules after detaching from stable Golgi structures. Brefeldin A treatment, which reversibly disassembles the Golgi complex, accentuated tubule formation without tubule detachment. A tubule network extending throughout the cytoplasm was quickly generated and persisted for 5-10 min until rapidly emptying Golgi contents into the ER within 15-30 s. Both lipid and protein emptied from the Golgi at similar rapid rates, leaving no Golgi structure behind, indicating that Golgi membranes do not simply mix but are absorbed into the ER in BFA-treated cells. The directionality of redistribution implied Golgi membranes are at a higher free energy state than ER membranes. Analysis of its kinetics suggested a mechanism that is analogous to wetting or adsorptive phenomena in which a tension-driven membrane flow supplements diffusive transfer of Golgi membrane into the ER. Such nonselective, flow-assisted transport of Golgi membranes into ER suggests that mechanisms that regulate retrograde tubule formation and detachment from the Golgi complex are integral to the existence and maintenance of this organelle.


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