Vacuolar protein sorting receptor in Schizosaccharomyces pombe

Abstract

The mechanism by which soluble proteins, such as carboxypeptidase Y, reach the vacuole inSaccharomyces cerevisiaeis very similar to the mechanism of lysosomal protein sorting in mammalian cells. Vps10p is a receptor for transport of soluble vacuolar proteins inS. cerevisiae.vps10+, a gene encoding a homologue ofS. cerevisiae PEP1/VPS10, has been identified and deleted from the fission yeastSchizosaccharomyces pombe. Deletion of thevps10+gene resulted in missorting and secretion ofSch. pombevacuolar carboxypeptidase Cpy1p, indicating that it is required for targeting Cpy1p to the vacuole.Sch. pombeVps10p (SpVps10p) is a type I transmembrane protein and its C-terminal cytoplasmic tail domain is essential for Cpy1p transport to the vacuole. Cells expressing green fluorescent protein-tagged SpVps10p produced a punctate pattern of fluorescence, indicating that SpVps10p was largely localized in the Golgi compartment. In addition,Sch. pombe vps26+,vps29+andvps35+, encoding homologues of theS. cerevisiaeretromer componentsVPS26,VPS29andVPS35, were identified and deleted. Fluorescence microscopy demonstrated that SpVps10p mislocalized to the vacuolar membrane in these mutants. These results indicate that thevps26+,vps29+andvps35+gene products are required for retrograde transport of SpVps10p from the prevacuolar compartment back to the Golgi inSch. pombecells.