Involvement of Protein N-Glycosyl Chain Glucosylation and Processing in the Biosynthesis of Cell Wall ß-1,6-Glucan of Saccharomyces cerevisiae

Corresponding author: Howard Bussey, Department of Biology, McGill University, 1205 Dr. Penfield Avenue, Montréal, QC, Canada H3A 1B1, hbussey{at}monod.biol.mcgill.ca (E-mail).

Communicating editor: M. JOHNSTON

ß-1,6-Glucan plays a key structural role in the yeast cell wall. Of the genes involved in its biosynthesis, the activity of Cwh41p is known, i.e., the glucosidase I enzyme of protein N-chain glucose processing. We therefore examined the effects of N-chain glucosylation and processing mutants on ß-1,6-glucan biosynthesis and show that incomplete N-chain glucose processing results in a loss of ß-1,6-glucan, demonstrating a relationship between N-chain glucosylation/processing and ß-1,6-glucan biosynthesis. To explore the involvement of other N-chain-dependent events with ß-1,6-glucan synthesis, we investigated the Saccharomyces cerevisiae KRE5 and CNE1 genes, which encode homologs of the "quality control" components UDP-Glc:glycoprotein glucosyltransferase and calnexin, respectively. We show that the essential activity of Kre5p is separate from its possible role as a UDP-Glc:glycoprotein glucosyltransferase. We also observe a ~30% decrease in ß-1,6-glucan upon disruption of the CNE1 gene, a phenotype that is additive with other ß-1,6-glucan synthetic mutants. Analysis of the cell wall anchorage of the mannoprotein -agglutinin suggests the existence of two ß-1,6-glucan biosynthetic pathways, one N-chain dependent, the other involving protein glycosylphosphatidylinositol modification.

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