Abstract
The S. cerevisiae VPS10 (vacuolar protein sorting) gene encodes a type I transmembrane protein of 1577 amino acids required for the sorting of the soluble vacuolar protein carboxypeptidase Y (CPY). Mutations in VPS10 result in the selective missorting and secretion of CPY; all other vacuolar proteins tested are delivered to the vacuole in vps10 mutants. Chemical cross-linking studies demonstrate that Vps10p and the Golgi-modified precursor form of CPY directly interact. A single amino acid change in the CPY vacuolar sorting signal prevents this interaction. Vps10p also interacts with a hybrid protein containing the CPY sorting signal fused to the normally secreted enzyme invertase. Subcellular fractionation indicates that the majority of Vps10p is localized to a late Golgi compartment where vacuolar proteins are sorted. We propose that VPS10 encodes a CPY sorting receptor that executes multiple rounds of sorting by cycling between the late Golgi and a prevacuolar endosome-like compartment.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Carboxypeptidases / analysis
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Carboxypeptidases / metabolism*
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Cathepsin A
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Cloning, Molecular
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Fungal Proteins / analysis
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Fungal Proteins / chemistry
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Fungal Proteins / genetics*
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Fungal Proteins / metabolism
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Genes, Fungal / genetics*
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Genetic Complementation Test
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Glycosylation
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Golgi Apparatus / chemistry
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Membrane Proteins / analysis
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Membrane Proteins / chemistry
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Membrane Proteins / genetics*
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Membrane Proteins / metabolism
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Models, Biological
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Molecular Sequence Data
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Mutation / physiology
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Receptors, Cell Surface / analysis
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Receptors, Cell Surface / chemistry
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Receptors, Cell Surface / genetics*
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Receptors, Cell Surface / metabolism
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Receptors, Cytoplasmic and Nuclear / analysis
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Receptors, Cytoplasmic and Nuclear / genetics*
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Receptors, Cytoplasmic and Nuclear / metabolism
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins*
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Sequence Analysis, DNA
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Sequence Homology, Amino Acid
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Spheroplasts / chemistry
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Vacuoles / chemistry
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Vacuoles / metabolism*
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Vesicular Transport Proteins*
Substances
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Fungal Proteins
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Membrane Proteins
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PEP1 protein, S cerevisiae
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Receptors, Cell Surface
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Receptors, Cytoplasmic and Nuclear
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Saccharomyces cerevisiae Proteins
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Vesicular Transport Proteins
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Carboxypeptidases
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Cathepsin A
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PRC1 protein, S cerevisiae
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serine carboxypeptidase