Isolation of temperature-sensitive Saccharomyces cerevisiae with a mutation in erg25 for C-4 sterol methyl oxidase.
C-4 sterol methyl oxidase encoded by the ERG25 gene is a key enzyme in the ergosterol biosynthetic pathway in fungi. ERG25p contains three histidine clusters common to nonheme iron binding enzymes and endoplasmic reticulum retrieval signal. In order to characterize ERG25p, we generated a series of temperature-sensitive(ts) erg25 mutants by random mutagenesis. One of the resulting mutants, the mERG25 strain, accumulated 4,4-dimethlzymosterol at the nonpermissive temperature. Sequence analysis of the mERG25 mutant indicated three amino acid substitutions in ERG25p, namely N48D, V133A, and F135S. These results indicate that the ERG25 gene product is a new antifungal target.
Nose
Hiroshi
H
Drug Discovery, Pharmaceutical Research Center, Meiji Seika Kaisha, Ltd., 760 Morooka-cho, Kohoku-ku, Yokohama 222-0002, Japan. hiroshi_nose@meiji.co.jp
Miyara
Takako
T
Kushida
Nobuaki
N
Hoshiko
Shigeru
S
D000595:Amino Acid Sequence; D002853:Chromatography, Liquid; D015870:Gene Expression; D006358:Hot Temperature; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D013261:Sterols / Q000032:analysis; D014162:Transfection; D014170:Transformation, Genetic
D013261:Sterols; D006899:Mixed Function Oxygenases; C027532:methylsterol monooxygenase