Isolation of the ALG6 locus of Saccharomyces cerevisiae required for glucosylation in the N-linked glycosylation pathway.
N-Linked protein glycosylation in most eukaryotic cells initiates with the transfer of the oligosaccharide Glc3Man9GlcNAc2 from the lipid carrier dolichyl pyrophosphate to selected asparagine residues. In the yeast Saccharomyces cerevisiae, alg mutations which affect the assembly of the lipid-linked oligosaccharide at the membrane of the endoplasmic reticulum result in the accumulation of lipid-linked oligosaccharide intermediates and a hypoglycosylation of proteins. Exploiting the synthetic growth defect of alg mutations in combination with mutations affecting oligosaccharyl transferase activity (Stagljar et al., 1994), we have isolated the ALG6 locus. alg6 mutants accumulate lipid-linked Man9GlcNAc2, suggesting that this locus encodes an endoplasmic glucosyltransferase. Alg6p has sequence similarity to Alg8p, a protein required for glucosylation of Glc1Man9GlcNAc2.
Reiss
G
G
Mikrobiologiches Institut, Eidgenössische Technische Hochschule, ETH Zentrum, Zürich, Switzerland.
te Heesen
S
S
Zimmerman
J
J
Robbins
P W
PW
Aebi
M
M
D000595:Amino Acid Sequence; D002240:Carbohydrate Sequence; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D005964:Glucosyltransferases / Q000235:genetics*; D008351:Mannans / Q000096:biosynthesis; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid
D005656:Fungal Proteins; D008351:Mannans; D029701:Saccharomyces cerevisiae Proteins; C057328:mannosyl(9)-N-acetylglucosamine; C087974:Alg8 protein, S cerevisiae; D005964:Glucosyltransferases