We have cloned the Saccharomyces cerevisiae C-4 sterol methyl oxidase ERG25 gene. The sterol methyl oxidase performs the first of three enzymic steps required to remove the two C-4 methyl groups leading to cholesterol (animal), ergosterol (fungal), and stigmasterol (plant) biosynthesis. An ergosterol auxotroph, erg25, which fails to demethylate and concomitantly accumulates 4,4-dimethylzy-mosterol, was isolated after mutagenesis. A complementing clone consisting of a 1.35-kb Dra I fragment encoded a 309-amino acid polypeptide (calculated molecular mass, 36.48 kDa). The amino acid sequence shows a C-terminal endoplasmic reticulum retrieval signal KKXX and three histidine-rich clusters found in eukaryotic membrane desaturases and in a bacterial alkane hydroxylase and xylene monooxygenase. The sterol profile of an ERG25 disruptant was consistent with the erg25 allele obtained by mutagenesis.
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Gene/Complex | Qualifier | Gene Ontology Term | Annotation Extension | Evidence | Source | Assigned On |
---|---|---|---|---|---|---|
ERG25 | enables | C-4 methylsterol oxidase activity | has input 14-demethyllanosterol, has output 4beta-methylzymosterol-4alpha-carboxylate, part of ergosterol biosynthetic process | IMP | UniProt | 2022-07-07 |
ERG25 | involved in | ergosterol biosynthetic process | IMP | UniProt | 2022-07-07 | |
ERG29 | acts upstream of or within | ergosterol biosynthetic process | IMP | UniProt | 2022-07-07 |
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Gene | Phenotype | Experiment Type | Mutant Information | Strain Background | Chemical | Details |
---|---|---|---|---|---|---|
ERG25 | auxotrophy | classical genetics | null Allele: erg25-Δ | Other | ergosterol | |
ERG25 | chemical compound accumulation: increased | classical genetics | null Allele: erg25-Δ | Other | 14-demethyllanosterol | Details: accumulation of 4,4-dimethylzymosterol and other C-4,4-dimethylsterols |