Identification of a Candida glabrata homologue of the S. cerevisiae VRG4 gene, encoding the Golgi GDP-mannose transporter

Mannoproteins on the cell wall of yeast and fungi help regulate cell shape, porosity, and cell-cell interactions, including those required for attachment to host cells by fungal pathogens. The mannose-containing oligosaccharides on proteins and lipids are extended in the Golgi by glycosyltransferases that use GDP-mannose as the sugar substrate. A membrane-bound transporter that, in Saccharomyces cerevisiae, is encoded by the VRG4 gene catalyses delivery of GDP-mannose into the lumen of the Golgi. We report here the cloning of the homologous VRG4 gene from the pathogenic yeast, Candida glabrata, by functional complementation of an S. cerevisiae vrg4 mutant. The sequence of the CgVrg4 protein displays significant homology to GDP-mannose transporters from other yeast, fungi, protozoa, and plants. CgVRG4 fully complements the glycosylation defect and other cell wall associated vrg4 mutant phenotypes. Like ScVRG4, CgVRG4 is essential for the viability of C. glabrata. These results suggest that, as in S. cerevisiae, CgVrg4p accounts for all of the GDP-mannose transport activity in the Golgi lumen. The GenBank Accession No. for CgVRG4 is #AF360395.