A nitrate-inducible ferredoxin in maize roots. Genomic organization and differential expression of two nonphotosynthetic ferredoxin isoproteins.
We have identified and characterized a nitrate-inducible ferredoxin (Fd) in maize (Zea mays L.) roots by structural analysis of the purified protein and by cloning of its cDNA and gene. In maize Fd isoproteins are encoded by a small multigene family, and the nitrate-inducible Fd was identified as a novel isoprotein, designated Fd VI, which differed from an Fd I to Fd V identified to date. In the roots of seedlings cultured without nitrate, Fd VI was undetectable. However, during the induction of the capacity for nitrate assimilation, the amount of Fd VI increased markedly within 24 h. Concurrently, the level of transcript for Fd VI increased, but more quickly, reaching a maximal level within 2 h with kinetics similar to those of nitrite reductase and Fd-NADP+ reductase. Fd III was constitutively expressed in roots, and no such changes at the protein and mRNA levels were observed during the nitrate induction. In the 5' flanking region of the gene for Fd VI only, we identified NIT-2 motifs, which are widely found in genes for enzymes related to nitrogen metabolism. These data indicate that Fd VI is co-induced with the previously characterized enzymes involved in nitrate assimilation, and they suggest that the novel Fd isoprotein, distinct from the constitutively expressed Fd, might play an important role as an electron carrier from NADPH to nitrite reductase and other Fd-dependent enzymes in root plastids.
Matsumura
T
T
Division of Enzymology, Osaksa University, Japan. matumura/biochem@nms.ac.jp
Sakakibara
H
H
Nakano
R
R
Kimata
Y
Y
Sugiyama
T
T
Hase
T
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D018076:DNA, Complementary / Q000235:genetics; D005287:Ferredoxin-NADP Reductase / Q000096:biosynthesis / Q000235:genetics; D005288:Ferredoxins / Q000096:biosynthesis / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D015723:Gene Library; D017343:Genes, Plant; D008969:Molecular Sequence Data; D009566:Nitrates / Q000378:metabolism / Q000494:pharmacology*; D009572:Nitrite Reductases / Q000096:biosynthesis / Q000235:genetics; D018517:Plant Roots / Q000187:drug effects / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid; D017422:Sequence Analysis, DNA; D003313:Zea mays / Q000187:drug effects / Q000235:genetics*
D018076:DNA, Complementary; D005288:Ferredoxins; D009566:Nitrates; D005287:Ferredoxin-NADP Reductase; D009572:Nitrite Reductases