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PMID: 7588624 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

STT3, a highly conserved protein required for yeast oligosaccharyl transferase activity in vivo.

The EMBO journal ·Vol. 14 ·No. 20 ·1995-10-16 ·Pages 4949-60

Zufferey R, Knauer R, Burda P, Stagljar I, te Heesen S, Lehle L, Aebi M

Abstract

N-linked glycosylation is a ubiquitous protein modification, and is essential for viability in eukaryotic cells. A lipid-linked core-oligosaccharide is assembled at the membrane of the endoplasmic reticulum and transferred to selected asparagine residues of nascent polypeptide chains by the oligosaccharyl transferase (OTase) complex. Based on the synthetic lethal phenotype of double mutations affecting the assembly of the lipid-linked core-oligosaccharide and the OTase activity, we have performed a novel screen for mutants in Saccharomyces cerevisiae with altered N-linked glycosylation. Besides novel mutants deficient in the assembly of the lipid-linked oligosaccharide (alg mutants), we identified the STT3 locus as being required for OTase activity in vivo. The essential STT3 protein is approximately 60% identical in amino acid sequence to its human homologue. A mutation in the STT3 locus affects substrate specificity of the OTase complex in vivo and in vitro. In stt3-3 cells very little glycosyl transfer occurs from incomplete lipid-linked oligosaccharide, whereas the transfer of full-length Glc3Man9GlcNAc2 is hardly affected as compared with wild-type cells. Depletion of the STT3 protein results in loss of transferase activity in vivo and a deficiency in the assembly of OTase complex.

MeSH Terms
Amino Acid Sequence Base Sequence Carbohydrate Sequence Conserved Sequence Fungal Proteins/genetics,metabolism Genes, Fungal Genes, Lethal Genetic Complementation Test Glycosylation Hexosyltransferases Membrane Proteins/genetics Molecular Sequence Data Mutation Protein Processing, Post-Translational/genetics Receptors, Cell Surface/metabolism Saccharomyces cerevisiae/enzymology,genetics Saccharomyces cerevisiae Proteins Selection, Genetic Sequence Homology, Amino Acid Substrate Specificity Transferases/genetics Vesicular Transport Proteins
Chemicals
Fungal Proteins Membrane Proteins PEP1 protein, S cerevisiae Receptors, Cell Surface Saccharomyces cerevisiae Proteins Vesicular Transport Proteins Transferases Hexosyltransferases STT3 protein, S cerevisiae dolichyl-diphosphooligosaccharide - protein glycotransferase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Zufferey R
Mikrobiologisches Institut, ETH Zürich, Switzerland.
Knauer R
Burda P
Stagljar I
te Heesen S
Lehle L
Aebi M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1995-10-16
Pages
4949-60
Language
English
Region
England
NLM ID
8208664
PMCID
PMC394598
Subset
IM
Databases
GENBANK
L38961, L43260, U13019
External Links
PubMed source PMC Full Text View journal details
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