Identification of Delta5-fatty acid desaturase from the cellular slime mold dictyostelium discoideum.
cDNA fragments putatively encoding amino acid sequences characteristic of the fatty acid desaturase were obtained using expressed sequence tag (EST) information of the Dictyostelium cDNA project. Using this sequence, we have determined the cDNA sequence and genomic sequence of a desaturase. The cloned cDNA is 1489 nucleotides long and the deduced amino acid sequence comprised 464 amino acid residues containing an N-terminal cytochrome b5 domain. The whole sequence was 38.6% identical to the initially identified Delta5-desaturase of Mortierella alpina. We have confirmed its function as Delta5-desaturase by over expression mutation in D. discoideum and also the gain of function mutation in the yeast Saccharomyces cerevisiae. Analysis of the lipids from transformed D. discoideum and yeast demonstrated the accumulation of Delta5-desaturated products. This is the first report concering fatty acid desaturase in cellular slime molds.
Saito
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Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo, Japan.
Ochiai
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H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D000090943:Delta-5 Fatty Acid Desaturase; D004023:Dictyostelium / Q000201:enzymology*; D044943:Fatty Acid Desaturases / Q000737:chemistry / Q000235:genetics*; D005227:Fatty Acids / Q000032:analysis; D015966:Gene Expression Regulation, Fungal; D008969:Molecular Sequence Data; D009154:Mutation; D012150:Polymorphism, Restriction Fragment Length; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment; D014170:Transformation, Genetic
D000090943:Delta-5 Fatty Acid Desaturase; D005227:Fatty Acids; D044943:Fatty Acid Desaturases