Arabidopsis alternative oxidase sustains Escherichia coli respiration.
Glutamyl-tRNA reductase, encoded by the hemA gene, is the first enzyme in porphyrin biosynthesis in many organisms. Hemes, important porphyrin derivatives, are essential components of redox enzymes, such as cytochromes. Thus a hemA Escherichia coli strain (SASX41B) is deficient in cytochrome-mediated aerobic respiration. Upon complementation of this strain with an Arabidopsis thaliana cDNA library, we isolated a clone which permitted the SASX41B strain to grow aerobically. The clone encodes the gene for Arabidopsis alternative oxidase, whose deduced amino acid sequence was found to have 71% identity with that of the enzyme from the voodoo lily, Sauromatum guttatum. The Arabidopsis protein is expressed as a 31-kDa protein in E. coli and confers on this organism cyanide-resistant growth, which in turn is sensitive to salicylhydroxamate. This implies that a single polypeptide is sufficient for alternative oxidase activity. Based on these observations we propose that a cyanide-insensitive respiratory pathway operates in the transformed E. coli hemA strain. Introduction of this pathway now opens the way to genetic/molecular biological investigations of alternative oxidase and its cofactor.
Kumar
A M
AM
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511.
Söll
D
D
D000445:Aldehyde Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D003486:Cyanides / Q000494:pharmacology; D003580:Cytochromes / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D015723:Gene Library; D017343:Genes, Plant; D005816:Genetic Complementation Test; D007700:Kinetics; D008954:Models, Biological; D008969:Molecular Sequence Data; D010101:Oxygen Consumption / Q000187:drug effects; D017386:Sequence Homology, Amino Acid
D003486:Cyanides; D003580:Cytochromes; D000445:Aldehyde Oxidoreductases; C067282:glutamyl tRNA reductase