Deletion of MCD 4 involved in glycosylphosphatidylinositol (GPI) anchor synthesis leads to an increase in beta-1,6-glucan level and a decrease in GPI-anchored protein and mannan levels in the cell wall of Saccharomyces cerevisiae.
Most proteins involved in the synthesis of the GPI core structure of Saccharomyces cerevisiae are essential for growth. To explore the relationship between the GPI anchor structure and beta-1,6-glucan synthesis, we screened deletion mutants in genes involved in GPI synthesis for osmotic remedial growth. Heterozygous diploid strains were dissected on medium with osmotic support and slow growth of the mcd 4 deletion mutant was observed. The mcd 4 mutant showed abnormal morphology and cell aggregation, and was hypersensitive to SDS, hygromycin B and K1 killer toxin. Incorporation of GPI cell wall proteins was examined using a GPI-Flo 1 fusion protein. The result suggested that the mcd 4 deletion causes a decrease in GPI cell wall proteins levels. The mutation also caused a decrease in mannan levels and an increase in alkali-insoluble beta-1,6-glucan and chitin levels in the cell wall.
Maneesri
Jaruwan
J
Department of Applied and Bioapplied Chemistry, Graduate School of Engineering, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan.
Azuma
Masayuki
M
Sakai
Yumiko
Y
Igarashi
Koichi
K
Matsumoto
Takeshi
T
Fukuda
Hideki
H
Kondo
Akihiko
A
Ooshima
Hiroshi
H
D002462:Cell Membrane / Q000378:metabolism*; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D008351:Mannans / Q000378:metabolism*; D008565:Membrane Proteins / Q000172:deficiency* / Q000235:genetics*; D016297:Mutagenesis, Site-Directed; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*; D013329:Structure-Activity Relationship; D014882:Water-Electrolyte Balance / Q000502:physiology; D047071:beta-Glucans / Q000378:metabolism*
D017261:Glycosylphosphatidylinositols; C118052:MCD4 protein, S cerevisiae; D008351:Mannans; D008565:Membrane Proteins; D029701:Saccharomyces cerevisiae Proteins; D047071:beta-Glucans; C064197:beta-1,6-glucan