ISOLATION OF THE ALG5 LOCUS ENCODING THE UDP-GLUCOSE-DOLICHYL-PHOSPHATE GLUCOSYLTRANSFERASE FROM SACCHAROMYCES-CEREVISIAE

UDP-glucose:dolichyl-phosphate glucosyltransferase is a transmembrane- bound enzyme of the endoplasmic reticulum involved in protein N-linked glycosylation. This enzyme catalyzes the transfer of glucose from UDP -glucose to dolichyl phosphate. The structural gene encoding this tran sferase from Saccharomyces cerevisiae was isolated by complementation of an alg5-1 mutation. DNA sequencing of ALG5 revealed an open-reading frame of 1002 bases encoding a transmembrane protein of molecular mas s 38.3 kDa. Overexpression of Alg5p in both yeast and Escherichia coli results in an increase of UDP-glucose:dolichyl-phosphate glucosyltran sferase activity, whereas a deletion of the yeast gene leads to a loss of this activity and a concomitant underglycosylation of carboxypepti dase Y. The ALG5 protein has sequence similarity to the GDP-mannose: d olichylphosphate mannosyltransferase (Dpm1p) from S. cerevisiae. Topol ogical studies indicate that UDP-glucose:dolichyl-phosphate glucosyltr ansferase is a transmembrane protein that spans the membrane several t imes.