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PMID: 1469044 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Yeast Kex1p is a Golgi-associated membrane protein: deletions in a cytoplasmic targeting domain result in mislocalization to the vacuolar membrane.

The Journal of cell biology ·Vol. 119 ·No. 6 ·1992-12-00 ·Pages 1459-68

Cooper A, Bussey H

Abstract

We have investigated the localization of Kex1p, a type I transmembrane carboxypeptidase involved in precursor processing within the yeast secretory pathway. Indirect immunofluorescence demonstrated the presence of Kex1p in a punctate organelle resembling the yeast Golgi apparatus as identified by Kex2p and Sec7p (Franzusoff, A., K. Redding, J. Crosby, R. S. Fuller, and R. Schekman. 1991. J. Cell Biol. 112:27-37). Glycosylation studies of Kex1p were consistent with a Golgi location, as Kex1p was progressively N-glycosylated in an MNN1-dependent manner. To address the basis of Kex1p targeting to the Golgi apparatus, we examined the cellular location of a series of carboxy-terminal truncations of the protein. The results indicate that a cytoplasmically exposed carboxy-terminal domain is required for retention of this membrane protein within the Golgi apparatus. Deletions of the retention region or overproduction of wild-type Kex1p led to mislocalization of Kex1p to the vacuolar membrane. This unexpected finding is discussed in terms of models involving either the vacuole as a default destination for membrane proteins, or by endocytosis to the vacuole following their default localization to the plasma membrane.

MeSH Terms
Amino Acid Sequence Base Sequence Biological Transport Carbohydrate Sequence Carboxypeptidases/genetics,isolation & purification,metabolism Cell Compartmentation DNA Mutational Analysis Fluorescent Antibody Technique Glycosylation Golgi Apparatus/metabolism Killer Factors, Yeast Models, Biological Molecular Sequence Data Mutagenesis, Site-Directed Mycotoxins/genetics,metabolism Protein Conformation Protein Processing, Post-Translational Recombinant Proteins Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Vacuoles/metabolism
Chemicals
KHR1 protein, S cerevisiae Killer Factors, Yeast Mycotoxins Recombinant Proteins Saccharomyces cerevisiae Proteins Carboxypeptidases carboxypeptidase D
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Cooper A
Department of Biology, McGill University, Montreal, Canada.
Bussey H
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.119.6.1459
Published
1992-12-00
Pages
1459-68
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289743
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.119.6.1459 PMC Full Text View journal details
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