N-glycosylation is involved in the sensitivity of Saccharomyces cerevisiae to HM-1 killer toxin secreted from Hansenula mrakii IFO 0895.
Saccharomyces cerevisiae rhk mutants were previously shown to have a phenotype that is resistant to HM-1 killer toxin secreted from Hansenula mrakii IFO 0895. The RHK1/ALG3 gene encodes a mannosyl-transferase that is involved in the synthesis of an oligosaccharide in protein N-glycosylation. Previously, this gene was cloned and shown to complement the rhk1 mutation. In this study, the RHK2 gene, which complements the rhk2 mutation, was cloned. The RHK2 gene was found to be identical to the essential gene STT3, which encodes a subunit of the oligosaccharyl-transferase complex. This complex transfers the core oligosaccharide to proteins. The rhk2 mutants showed supersensitivity to several drugs (Calcofluor White, caffeine and FK506), suggesting that these strains have cell-wall defects. Activity staining of invertase in an acrylamide gel indicated that it was underglycosylated. These results suggest that one or more mannoproteins are involved in the cytocidal process of HM-1.
Kimura
T
T
Faculty of Bioresources, Mie University, Japan. t-kimura@bio.mie-u.ac.jp
Komiyama
T
T
Furuichi
Y
Y
Iimura
Y
Y
Karita
S
S
Sakka
K
K
Ohmiya
K
K
D001557:Benzenesulfonates / Q000494:pharmacology; D002110:Caffeine / Q000494:pharmacology; D004352:Drug Resistance, Microbial / Q000235:genetics; D005456:Fluorescent Dyes / Q000494:pharmacology; D005800:Genes, Fungal; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D006602:Hexosyltransferases; D007166:Immunosuppressive Agents / Q000494:pharmacology; D055646:Killer Factors, Yeast; D008565:Membrane Proteins / Q000235:genetics; D008826:Microbial Sensitivity Tests; D009154:Mutation; D009183:Mycotoxins / Q000378:metabolism* / Q000494:pharmacology*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010843:Pichia / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D016559:Tacrolimus / Q000494:pharmacology; D014166:Transferases / Q000235:genetics; D043324:beta-Fructofuranosidase
D001557:Benzenesulfonates; D005456:Fluorescent Dyes; D007166:Immunosuppressive Agents; D055646:Killer Factors, Yeast; D008565:Membrane Proteins; D009183:Mycotoxins; D010726:Phosphodiesterase Inhibitors; D029701:Saccharomyces cerevisiae Proteins; D002110:Caffeine; C007061:C.I. Fluorescent Brightening Agent 28; D014166:Transferases; D006602:Hexosyltransferases; C096554:STT3 protein, S cerevisiae; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D016559:Tacrolimus