The OLE1 gene of Saccharomyces cerevisiae encodes the delta 9 fatty acid desaturase and can be functionally replaced by the rat stearoyl-CoA desaturase gene.
Strains of Saccharomyces cerevisiae bearing the ole1 mutation are defective in unsaturated fatty acid (UFA) synthesis and require UFAs for growth. A previously isolated yeast genomic fragment complementing the ole1 mutation has been sequenced and determined to encode the delta 9 fatty acid desaturase enzyme by comparison of primary amino acid sequence to the rat liver stearoyl-CoA desaturase. The OLE1 structural gene encodes a protein of 510 amino acids (251 hydrophobic) having an approximate molecular mass of 57.4 kDa. A 257-amino acid internal region of the yeast open reading frame aligns with and shows 36% identity and 60% similarity to the rat liver stearoyl-CoA desaturase protein. This comparison disclosed three short regions of high consecutive amino acid identity (greater than 70%) including one 11 of 12 perfect residue match. The predicted yeast enzyme contains at least four potential membrane-spanning regions and several shorter hydrophobic regions that align exactly with similar sequences in the rat liver protein. An ole1 gene-disrupted yeast strain was transformed with a yeast-rat chimeric gene consisting of the promoter region and N-terminal 27 codons of OLE1 fused to the rat desaturase coding sequence. Fusion gene transformants displayed near equivalent growth rates and modest lipid composition changes relative to wild type yeast control implying a significant conservation of delta 9 desaturase tertiary structure and efficient interaction between the rat desaturase and yeast cytochrome b5.
Stukey
J E
JE
Nelson Biological Laboratory, Rutgers University, Piscataway, New Jersey 08855-1059.
McDonough
V M
VM
Martin
C E
CE
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004271:DNA, Fungal / Q000235:genetics; D004721:Endoplasmic Reticulum / Q000201:enzymology / Q000648:ultrastructure; D044943:Fatty Acid Desaturases / Q000235:genetics*; D005796:Genes; D005800:Genes, Fungal; D008961:Models, Structural; D008969:Molecular Sequence Data; D011487:Protein Conformation; D051381:Rats; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D012689:Sequence Homology, Nucleic Acid; D013230:Stearoyl-CoA Desaturase / Q000235:genetics*; D016385:TATA Box
D004271:DNA, Fungal; D044943:Fatty Acid Desaturases; D013230:Stearoyl-CoA Desaturase