title: Salt shock-inducible photosystem I cyclic electron transfer in Synechocystis PCC6803 relies on binding of ferredoxin:NADP(+) reductase to the thylakoid membranes via its CpcD phycobilisome-linker homologous N-terminal domain.
abstract: Relative to ferredoxin:NADP(+) reductase (FNR) from chloroplasts, the comparable enzyme in cyanobacteria contains an additional 9 kDa domain at its amino-terminus. The domain is homologous to the phycocyanin associated linker polypeptide CpcD of the light harvesting phycobilisome antennae. The phenotypic consequences of the genetic removal of this domain from the petH gene, which encodes FNR, have been studied in Synechocystis PCC 6803. The in frame deletion of 75 residues at the amino-terminus, rendered chloroplast length FNR enzyme with normal functionality in linear photosynthetic electron transfer. Salt shock correlated with increased abundance of petH mRNA in the wild-type and mutant alike. The truncation stopped salt stress-inducible increase of Photosystem I-dependent cyclic electron flow. Both photoacoustic determination of the storage of energy from Photosystem I specific far-red light, and the re-reduction kinetics of P700(+), suggest lack of function of the truncated FNR in the plastoquinone-cytochrome b(6)f complex reductase step of the PS I-dependent cyclic electron transfer chain. Independent gold-immunodecoration studies and analysis of FNR distribution through activity staining after native polyacrylamide gelelectrophoresis showed that association of FNR with the thylakoid membranes of Synechocystis PCC 6803 requires the presence of the extended amino-terminal domain of the enzyme. The truncated DeltapetH gene was also transformed into a NAD(P)H dehydrogenase (NDH1) deficient mutant of Synechocystis PCC 6803 (strain M55) (T. Ogawa, Proc. Natl. Acad. Sci. USA 88 (1991) 4275-4279). Phenotypic characterisation of the double mutant supported our conclusion that both the NAD(P)H dehydrogenase complex and FNR contribute independently to the quinone cytochrome b(6)f reductase step in PS I-dependent cyclic electron transfer. The distribution, binding properties and function of FNR in the model cyanobacterium Synechocystis PCC 6803 will be discussed.
authors.affiliation: E.C. Slater Institute, University of Amsterdam, Nieuwe Achtergracht 127, 1018 WS, Amsterdam, The Netherlands.
authors.forename: J J
authors.initials: JJ
authors.lastname: van Thor
authors.forename: R
authors.initials: R
authors.lastname: Jeanjean
authors.forename: M
authors.initials: M
authors.lastname: Havaux
authors.forename: K A
authors.initials: KA
authors.lastname: Sjollema
authors.forename: F
authors.initials: F
authors.lastname: Joset
authors.forename: K J
authors.initials: KJ
authors.lastname: Hellingwerf
authors.forename: H C
authors.initials: HC
authors.lastname: Matthijs
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D001665:Binding Sites; D002021:Buffers; D000458:Cyanobacteria / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D004579:Electron Transport; D005287:Ferredoxin-NADP Reductase / Q000737:chemistry* / Q000235:genetics; D005420:Flavoproteins; D049452:Green Fluorescent Proteins; D045342:Light-Harvesting Protein Complexes; D008164:Luminescent Proteins / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D045524:Phycobilisomes; D012333:RNA, Messenger / Q000096:biosynthesis; D012965:Sodium Chloride; D020524:Thylakoids / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D002021:Buffers; D005420:Flavoproteins; D045342:Light-Harvesting Protein Complexes; D008164:Luminescent Proteins; D008565:Membrane Proteins; D045322:Photosynthetic Reaction Center Complex Proteins; D045524:Phycobilisomes; D012333:RNA, Messenger; C082501:cpcD phycobilisome linker protein, cyanobacteria; D049452:Green Fluorescent Proteins; D012965:Sodium Chloride; D005287:Ferredoxin-NADP Reductase; C082502:petH protein, Bacteria