An Arabidopsis gene induced by wounding functionally homologous to flavoprotein oxidoreductases.
The regulation of genes in response to wounding is mediated in part by the octadecanoids 12-oxo-phytodienoic acid (OPDA), jasmonic acid (JA) and its methyl ester methyl jasmonate (MeJA). We identified, by differential display, an Arabidopsis gene (OPR3) induced after wounding. OPR3 is homologous to members of the flavin mononucleotide (FMN) binding proteins, including the old yellow enzyme (OYE) from yeast and 12-oxophytodienoate-10,11-reductase (OPR) from Arabidopsis. Transcripts of OPR3 rapidly accumulated in leaves after wounding and MeJA treatment, but they were detected in various tissues of unwounded plants at relatively low levels. Expression of the OPR3 gene was significantly reduced in wounded leaves of the coil mutant, indicating partial dependence on jasmonate perception for full induction of the gene. The recombinant protein of OPR3 cross-reacted with an antiserum raised against the OYE protein, and showed oxidation of beta-NADPH when OPDA or 15-deoxy-delta(12,14) prostaglandin J2 (PGJ2), an analogue of OPDA, was used as substrate. Beta-NADPH oxidation was not observed when MeJA, which lacks the double bond in the ketone ring, was used as substrate. The recombinant OPR3 protein also showed beta-NADPH oxidation activity in the presence of cyclohexenone, but not cyclohexanone, suggesting that the enzyme has specificity to cleavage of olefinic bonds in cyclic enones. The results show that the OPR3 gene product represents a new OPR of Arabidopsis induced after wounding.
Costa
C L
CL
Centro de Biologia Molecular e Engenharia Genética, Universidade Estadual de Campinas, SP, Brazil.
Arruda
P
P
Benedetti
C E
CE
D000085:Acetates / Q000494:pharmacology; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins; D015152:Blotting, Northern; D003517:Cyclopentanes / Q000494:pharmacology; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D005420:Flavoproteins; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D017343:Genes, Plant / Q000235:genetics*; D007700:Kinetics; D008969:Molecular Sequence Data; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000235:genetics*; D054883:Oxylipins; D010940:Plant Proteins; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013314:Stress, Mechanical; D013379:Substrate Specificity; D014018:Tissue Distribution
D000085:Acetates; D029681:Arabidopsis Proteins; D003517:Cyclopentanes; D018076:DNA, Complementary; D005420:Flavoproteins; D054883:Oxylipins; D010940:Plant Proteins; D018749:RNA, Plant; D011994:Recombinant Proteins; D009249:NADP; C072239:methyl jasmonate; C416525:OPR3 protein, Arabidopsis; D010088:Oxidoreductases; D009247:NADH, NADPH Oxidoreductases