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PMID: 3047151 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

A membrane glycoprotein, Sec12p, required for protein transport from the endoplasmic reticulum to the Golgi apparatus in yeast.

The Journal of cell biology ·Vol. 107 ·No. 3 ·1988-09-00 ·Pages 851-63

Nakano A, Brada D, Schekman R

Abstract

SEC12, a gene that is required for secretory, membrane, and vacuolar proteins to be transported from the endoplasmic reticulum to the Golgi apparatus, has been cloned from a genomic library by complementation of a sec12 ts mutation. Genetic analysis has shown that the cloned gene integrates at the SEC12 locus and that a null mutation at the locus is lethal. The DNA sequence predicts a protein of 471 amino acids containing a hydrophobic stretch of 19 amino acids near the COOH terminus. To characterize the gene product (Sec12p) in detail, a lacZ-SEC12 gene fusion has been constructed and a polyclonal antibody raised against the hybrid protein. The antibody recognizes Sec12p as a approximately 70-kD protein that sediments in a mixed membrane fraction that includes endoplasmic reticulum. Sec12p is not removed from the membrane fraction by treatment at high pH and high salt and is not degraded by exogenous protease unless detergent is present. Glycosylation of Sec12p during biogenesis is indicated by an electrophoretic mobility shift of the protein that is influenced by tunicamycin and by imposition of an independent secretory pathway block. We suggest that Sec12p is an integral membrane glycoprotein with a prominent domain that faces the cytoplasm where it functions to promote protein transport to the Golgi apparatus. In the process of transport, Sec12p itself may migrate to the Golgi apparatus and function in subsequent transport events.

MeSH Terms
Amino Acid Sequence Base Sequence Cloning, Molecular DNA, Fungal/genetics Endoplasmic Reticulum/metabolism Fungal Proteins/metabolism Genes, Fungal Genetic Complementation Test Glycosylation Golgi Apparatus/metabolism Membrane Glycoproteins/genetics,metabolism Microscopy, Electron Molecular Sequence Data Mutation Nucleic Acid Hybridization Plasmids Protein Processing, Post-Translational RNA, Fungal/analysis Saccharomyces cerevisiae/genetics,metabolism,ultrastructure Transcription, Genetic
Chemicals
DNA, Fungal Fungal Proteins Membrane Glycoproteins RNA, Fungal
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Nakano A
Department of Biochemistry, University of California, Berkeley 94720.
Brada D
Schekman R
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38 references, click to expand
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.107.3.851
Published
1988-09-00
Pages
851-63
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2115266
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.107.3.851 PMC Full Text View journal details
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