The modification of yeast uracil permease by phosphorylation at the plasma membrane is a key mechanism for regulating transporter endocytosis. Uracil permease is phosphorylated at several serine residues within a well characterized PEST sequence. The phosphorylation of these residues facilitates the ubiquitination and internalization of the permease. Following endocytosis, the permease is targeted to the lysosome/vacuole for proteolysis. We have shown that in casein kinase 1 (CK1)-deficient cells, the permease is poorly phosphorylated, poorly ubiquitinated and that Yck activity may play a direct role in phosphorylating the permease. We show here that CK1-deficient cells accumulated permease that was subjected to endocytosis in an internal compartment on its way to the vacuole. Uracil permease, produced as a fusion protein with green fluorescent protein in CK1-deficient cells, was detected in dots adjacent to the vacuole. These dots probably correspond to the late endosome/prevacuolar compartment because they were partially colocalized with the Pep12p marker. This accumulation was abolished by mutations affecting the adaptor-related complex, AP-3. The CPY and ALP pathways to the vacuole were both unaffected in CK1-deficient cells. Our analysis provides the first evidence that CK1 is important for the delivery of proteins to the vacuole after endocytosis.

• PMID: 11801739
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Journal Article | Research Support, Non-U.S. Gov't

Authors

Marchal C, Dupré S, Urban-Grimal D

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