Molecular cloning and functional identification of a plant ornithine decarboxylase cDNA.
A cDNA for a plant ornithine decarboxylase (ODC), a key enzyme in putrescine and polyamine biosynthesis, has been isolated from root cultures of the solanaceous plant Datura stramonium. Reverse transcription-PCR employing degenerate oligonucleotide primers representing conserved motifs from other eukaryotic ODCs was used to isolate the cDNA. The longest open reading frame potentially encodes a peptide of 431 amino acids and exhibits similarity to other eukaryotic ODCs, prokaryotic and eukaryotic arginine decarboxylases (ADCs), prokaryotic meso-diaminopimelate decarboxylases and the product of the tabA gene of Pseudomonas syringae cv. tabaci. Residues involved at the active site of the mouse ODC are conserved in the plant enzyme. The plant ODC does not possess the C-terminal extension found in the mammalian enzyme, implicated in rapid turnover of the protein, suggesting that the plant ODC may have a longer half-life. Expression of the plant ODC in Escherichia coli and demonstration of ODC activity confirmed that the cDNA encodes an active ODC enzyme. This is the first description of the primary structure of a eukaryotic ODC isolated from an organism where the alternative ADC routine to putrescine is present.
Michael
A J
AJ
Department of Genetics and Microbiology, Institute of Food Research, Norwich, U.K.
Furze
J M
JM
Rhodes
M J
MJ
Burtin
D
D
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002262:Carboxy-Lyases / Q000235:genetics; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics*; D013286:Datura stramonium / Q000201:enzymology* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D019143:Evolution, Molecular; D018628:Gene Dosage; D015870:Gene Expression; D017343:Genes, Plant; D006801:Humans; D008969:Molecular Sequence Data; D009955:Ornithine Decarboxylase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010946:Plants, Medicinal; D010947:Plants, Toxic; D011073:Polyamines / Q000378:metabolism; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D014158:Transcription, Genetic / Q000235:genetics
D018076:DNA, Complementary; D018744:DNA, Plant; D011073:Polyamines; D002262:Carboxy-Lyases; D009955:Ornithine Decarboxylase; C019722:arginine decarboxylase