Raft partitioning of the yeast uracil permease during trafficking along the endocytic pathway.
Lipid rafts, formed by the lateral association of sphingolipids and cholesterol in the external membrane leaflet, have been implicated in membrane traffic and cell signaling in mammalian cells. Yeast plasma membranes were also recently shown to contain lipid raft microdomains consisting of sphingolipids and ergosterol, and containing several plasma membrane proteins, including Gas1p, a GPI-anchored protein, and the [H+] ATPase Pma1p. In this study, we investigated whether lipid rafts were involved in the intracellular trafficking of a yeast transporter, uracil permease, which undergoes ubiquitin-dependent endocytosis. Regardless of its ubiquitination status, uracil permease was found to be associated with rafts in the plasma membrane. The expression of Fur4p in lcb1-100 cells, deficient in the first enzyme of sphingolipid synthesis, impaired the association of Fur4p with detergent-resistant fractions. When targeted to endocytic compartments, uracil permease appeared to be progressively transferred to detergent-soluble fractions, suggesting that the lipid environment might change between plasma membrane and endosomes. Consistent with this hypothesis, the wild-type form of the v-SNARE Snc1p, which is known to cycle between the plasma membrane and endosomal compartments, was recovered in both detergent-resistant and detergent-soluble fractions. In contrast, a variant Snc1p that accumulates at the plasma membrane was recovered exclusively in detergent-resistant fractions.
Dupré
Sophie
S
Institut Jacques Monod-CNRS, Universités Paris VI and Paris VII, 2 place Jussieu, 75005 Paris, France.
Haguenauer-Tsapis
Rosine
R
D003902:Detergents / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology*; D017930:Genes, Reporter; D021962:Membrane Microdomains / Q000378:metabolism*; D009154:Mutation; D033704:Nucleotide Transport Proteins / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
D003902:Detergents; C035528:FUR4 protein, S cerevisiae; D033704:Nucleotide Transport Proteins; D029701:Saccharomyces cerevisiae Proteins