A second isoform of the ferredoxin:NADP oxidoreductase generated by an in-frame initiation of translation.
Ferredoxin:NADP oxidoreductases (FNRs) constitute a family of flavoenzymes that catalyze the exchange of reducing equivalents between one-electron carriers and the two-electron-carrying NADP(H). The main role of FNRs in cyanobacteria and leaf plastids is to provide the NADPH for photoautotrophic metabolism. In root plastids, a distinct FNR isoform is found that has been postulated to function in the opposite direction, providing electrons for nitrogen assimilation at the expense of NADPH generated by heterotrophic metabolism. A multiple gene family encodes FNR isoenzymes in plants, whereas there is only one FNR gene (petH) in cyanobacteria. Nevertheless, we detected two FNR isoforms in the cyanobacterium Synechocystis sp. strain PCC6803. One of them (FNR(S) approximately 34 kDa) is similar in size to the plastid FNR and specifically accumulates under heterotrophic conditions, whereas the other one (FNR(L) approximately 46 kDa) contains an extra N-terminal domain that allows its association with the phycobilisome. Site-directed mutants allowed us to conclude that the smaller isoform, FNR(S), is produced from an internal ribosome entry site within the petH ORF. Thus we have uncovered a mechanism by which two isoforms are produced from a single gene, which is, to our knowledge, novel in photosynthetic bacteria. Our results strongly suggest that FNR(L) is an NADP(+) reductase, whereas FNR(S) is an NADPH oxidase.
Thomas
Jean-Claude
JC
Département de Biologie, Ecole Normale Supérieure, F-75230 Paris, France.
Ughy
Bettina
B
Lagoutte
Bernard
B
Ajlani
Ghada
G
D000595:Amino Acid Sequence; D001483:Base Sequence; D002457:Cell Extracts; D018387:Codon, Initiator / Q000235:genetics*; D005287:Ferredoxin-NADP Reductase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008715:Methionine / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010442:Peptide Chain Initiation, Translational / Q000235:genetics*; D045524:Phycobilisomes / Q000235:genetics; D016415:Sequence Alignment; D046939:Synechocystis / Q000201:enzymology* / Q000235:genetics*
D002457:Cell Extracts; D018387:Codon, Initiator; D007527:Isoenzymes; D045524:Phycobilisomes; D008715:Methionine; D005287:Ferredoxin-NADP Reductase