Differential induction of pyruvate decarboxylase subunits and transcripts in anoxic rice seedlings.
In 2-d-old rice (Oryza sativa L.) seedlings subjected to anoxic stress, pyruvate decarboxylase (PDC) activity increased 9-fold during a 168-h period. A polyclonal PDC antiserum that recognized alpha- and beta-subunits was used to quantify PDC protein by an enzyme-linked immunosorbant assay and showed a 5.6-fold increase, suggesting that the anoxically induced enzyme has a higher specific activity than the PDC isoform present under normoxia. Immunoblot analysis showed that levels of both PDC subunits were induced by anoxia. Immunoprecipitation of proteins labeled in vivo during anoxic treatment demonstrated that the alpha-subunit was preferentially synthesized at the onset of anoxia. Two partial cDNAs, including a novel sequence, were cloned from a cDNA library made from seedlings subjected to anoxia for 6 h. Gene-specific probes used to quantify northern blots showed that two or three PDC mRNAs are differentially induced by anoxia in rice seedlings. Immunoprecipitation of in vitro translation products of mRNAs isolated a different times of anoxic treatment confirmed this findings Our results suggest that anoxic induction of rice PDC involves transcriptional and posttranscriptional regulation of gene expression as well as differences in enzyme characteristics.
Rivoal
J
J
Station de Physiologie Végétale, Institut Nationale de la Recherche Agronomique, Centre de Recherches de Bordeaux, Villenave d'Ornon, France.
Thind
S
S
Pradet
A
A
Ricard
B
B
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004790:Enzyme Induction; D018506:Gene Expression Regulation, Plant; D000860:Hypoxia; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology* / Q000254:growth & development; D018515:Plant Leaves; D011767:Pyruvate Decarboxylase / Q000096:biosynthesis* / Q000737:chemistry; D012323:RNA Processing, Post-Transcriptional; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic
D018076:DNA, Complementary; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D011767:Pyruvate Decarboxylase