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MBC in Press, published online ahead of print June 6, 2007
Mol. Biol. Cell 10.1091/mbc.E07-03-0196

A more recent version of this article appeared on August 1, 2007
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Submitted on March 2, 2007
Revised on May 18, 2007
Accepted on May 29, 2007

Evidence for Coupled Biogenesis of Yeast Gap1 Permease and Sphingolipids: Essential Role in Transport Activity and Normal Control by Ubiquitination

Elsa Lauwers,* Guido Grossmann,{dagger} and Bruno André*

*Laboratoire de Physiologie Moléculaire de la Cellule, Institut de Biologie et de Médecine Moléculaires, Université Libre de Bruxelles, B-6041 Gosselies, Belgium; {dagger}University of Regensburg, Cell Biology and Plant Physiology, 93040 Regensburg, Germany

Monitoring Editor: Thomas Sommer

Current models for plasma membrane organization integrate the emerging concepts that membrane proteins tightly associate with surrounding lipids and that biogenesis of surface proteins and lipids may be coupled. We show here that the yeast general amino acid permease Gap1 synthesized in the absence of sphingolipid (SL) biosynthesis is delivered to the cell surface but undergoes rapid and unregulated down-regulation. Furthermore, the permease produced under these conditions but blocked at the cell surface is inactive, soluble in detergent, and more sensitive to proteases. We also show that SL biogenesis is crucial during Gap1 production and secretion but dispensable once Gap1 has reached the plasma membrane. Moreover, the defects displayed by cell surface Gap1 neosynthesized in the absence of SL biosynthesis are not compensated by subsequent restoration of SL production. Finally, we show that down-regulation of Gap1 caused by lack of SL biogenesis involves the ubiquitination of the protein on lysines normally not accessible to ubiquitination and close to the membrane. We propose that coupled biogenesis of Gap1 and SLs would create a SL microenvironment essential to the normal conformation, function and control of ubiquitination of the permease.

Address correspondence to: Bruno André (bran{at}ulb.ac.be)




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R. C. Dickson
Thematic Review Series: Sphingolipids. New insights into sphingolipid metabolism and function in budding yeast
J. Lipid Res., May 1, 2008; 49(5): 909 - 921.
[Abstract] [Full Text] [PDF]


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