KEG1/YFR042w encodes a novel Kre6-binding endoplasmic reticulum membrane protein responsible for β-1,6-glucan synthesis in Saccharomyces cerevisiae

Abstract

KEG1/YFR042w of Saccharomyces cerevisiae is an essential gene that encodes a 200-amino acid polypeptide with four predicted transmembrane domains. The green fluorescent protein- or Myc6-tagged Keg1 protein showed the typical characteristics of an integral membrane protein and was found in the endoplasmic reticulum by fluorescence imaging. Immunoprecipitation from the Triton X-100-solubilized cell lysate revealed that Keg1 binds to Kre6, which has been known to participate in β-1,6-glucan synthesis. To analyze the essential function of Keg1 in more detail, we constructed temperature-sensitive mutant alleles by error-prone polymerase chain reaction. The keg1-1 mutant cells showed a common phenotype with Δkre6 mutant including hypersensitivity to Calcofluor white, reduced sensitivity to the K1 killer toxin, and reduced content of β-1,6-glucan in the cell wall. These results suggest that Keg1 and Kre6 have a cooperative role in β-1,6-glucan synthesis in S. cerevisiae. © 2007 by The American Society for Biochemistry and Molecular Biology, Inc.