Cloning and expression in Escherichia coli of a yeast mannosyltransferase from the asparagine-linked glycosylation pathway

The yeast Saccharomyces cerevisiae temperature-sensitive lethal mutant alg1-1, has been previously shown to lack the activity necessary for the addition of the first mannose residue in the synthesis of lipid-linked precursor oligosaccharide. The gene ALG1 has been cloned by complementation of the te...

Full description

Saved in:
Bibliographic Details
Published in:The Journal of biological chemistry 1984-01, Vol.259 (1), p.378-382
Main Authors: Couto, J R, Huffaker, T C, Robbins, P W
Format: Article
Language:English
Subjects:
Asparagine
Biological and medical sciences
Biotechnology
Chromosome Mapping
Cloning, Molecular
Escherichia coli - genetics
Fundamental and applied biological sciences. Psychology
Genes
Genetic engineering
Genetic technics
Hexosyltransferases - genetics
Mannosyltransferases - genetics
Methods. Procedures. Technologies
Molecular cloning
Mutation
Plasmids
Saccharomyces cerevisiae - enzymology
Saccharomyces cerevisiae - genetics
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The yeast Saccharomyces cerevisiae temperature-sensitive lethal mutant alg1-1, has been previously shown to lack the activity necessary for the addition of the first mannose residue in the synthesis of lipid-linked precursor oligosaccharide. The gene ALG1 has been cloned by complementation of the temperature-sensitive mutation alg1-1 with a total genomic DNA library. The original DNA fragment isolated was 11,300 base pairs and has been subcloned to a 1,500-base pair fragment which is still capable of complementing alg1-1. The gene ALG1 has been mapped on chromosome II at a distance of 2.1 map units from LYS2. The ALG1 gene product has been shown to catalyze the transfer of a mannosyl residue from GDP-mannose to the lipid-linked acceptor GlcNAc2, yielding Man beta 1-4GlcNAc2-lipid, in lysates from Escherichia coli transformants. This result proves that ALG1 is the structural gene for the first mannosyltransferase in lipid-linked oligosaccharide assembly.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(17)43670-2