Functional identification of sterol-4alpha-methyl oxidase cDNAs from Arabidopsis thaliana by complementation of a yeast erg25 mutant lacking sterol-4alpha-methyl oxidation.
Specific primers derived from both genomic sequence data and EST cDNA sequences were used to polymerase chain reaction amplify two full-length cDNA sequences (AtSMO1 and AtSMO2), 801 and 783 bp, respectively, from an Arabidopsis thaliana cDNA library. The predicted proteins show 32 and 29% identity to the ERG25 gene from Saccharomyces cerevisiae which encodes the sterol-4alpha-methyl oxidase (SMO), a membrane-bound non-heme di-iron oxygenase involved in lipid metabolism. Heterologous expression of AtSMO1 and AtSMO2 in a yeast erg25 ergosterol auxotroph, lacking SMO activity, restored growth and endogenous ergosterol synthesis. These results represent the first functional identification of SMO genes from plants.
Darnet
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Département Isoprénoïdes, Institut de Biologie Moléculaire des Plantes, UPR-CNRS 2357, 28 rue Goethe, 67083 Strasbourg Cedex, France.
Bard
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Rahier
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D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D002849:Chromatography, Gas; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004875:Ergosterol / Q000096:biosynthesis; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009154:Mutation; D010084:Oxidation-Reduction; D010802:Phylogeny; D010840:Phytosterols / Q000737:chemistry / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics
D029681:Arabidopsis Proteins; D018076:DNA, Complementary; D007527:Isoenzymes; D010840:Phytosterols; D006899:Mixed Function Oxygenases; C027532:methylsterol monooxygenase; D004875:Ergosterol