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Published online 3 December 2001. doi:10.1083/jcb.200105033
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© The Rockefeller University Press, 0021-9525/2001/12/949 $5.00
The Journal of Cell Biology, Volume 155, Number 6, December 10, 2001 949-960

Article

Vesicular and nonvesicular transport of ceramide from ER to the Golgi apparatus in yeast



Kouichi Funato and Howard Riezman

Biozentrum of the University of Basel, CH-4056 Basel, Switzerland

Address correspondence to Howard Riezman, Biozentrum, University of Basel, Klingelbergstr. 70, Basel CH-4056, Switzerland. Tel.: 41-61-267-2160. Fax: 41-61-267-2149. E-mail: howard.riezman{at}unibas.ch

Transport and sorting of lipids must occur with specific mechanisms because the membranes of intracellular organelles differ in lipid composition even though most lipid biosynthesis begins in the ER. In yeast, ceramide is synthesized in the ER and transferred to the Golgi apparatus where inositolphosphorylceramide (IPC) is formed. These two facts imply that ceramide can be transported to the Golgi independent of vesicular traffic because IPC synthesis still continues when vesicular transport is blocked in sec mutants. Nonvesicular IPC synthesis in intact cells is not affected by ATP depletion. Using an in vitro assay that reconstitutes the nonvesicular pathway for transport of ceramide, we found that transport is temperature and cytosol dependent but energy independent. Preincubation of ER and Golgi fractions together at 4°C, where ceramide transport does not occur, rendered the transport reaction membrane concentration independent, providing biochemical evidence that ER-Golgi membrane contacts stimulate ceramide transport. A cytosolic protease-sensitive factor is required after establishment of ER-Golgi contacts.

Key Words: ceramide transport; sphingolipids; membrane contacts; secretion; Saccharomyces cerevisiae


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