Function and organization of photosystem I in a cyanobacterial mutant strain that lacks PsaF and PsaJ subunits.
Photosystem I functions as a light-driven plastocyanin-ferredoxin oxidoreductase in the photosynthetic membranes of cyanobacteria and chloroplasts. A mutant strain of the cyanobacterium Synechocystis sp. PCC 6803 that contains a deletion of the psaF gene and a transcriptionally inactive psaJ gene has assembled photosystem I complexes that lack PsaF, a lumenal protein and PsaJ, a 4-kDa hydrophobic protein. The cells of the mutant and wild type strains have similar rates of photosynthetic electron transfer and P700+ rereduction under linear and cyclic electron transfer conditions. Analysis of flash-induced absorption transients at 700 nm demonstrate that the absence of PsaF in purified mutant photosystem I did not affect the rate of P700 rereduction by cytochrome c553. Therefore, PsaF is not essential for docking of cytochrome c553. We also studied the organization of the proteins of mutant and wild type photosystem I by comparing their accessibility to digestion by thermolysin or to removal by 1 M NaI. The PsaA-PsaB subunits were more easily degraded by thermolysin in the mutant photosystem I. Thermolysin cleavage of PsaB yielded two major fragments that were immunoreactive with an antibody raised against the C terminus of PsaB. The N termini of these PsaB peptides mapped at Ile482 and Ile498 residues, thus identifying a surface-exposed domain of the core of photosystem I. The PsaE subunit could be removed by 1 M NaI and was rapidly digested by thermolysin in the mutant but not in the wild type photosystem I. Therefore, PsaF and PsaJ subunits of photosystem I have dispensable accessory roles in the function and organization of the complex.
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Division of Biology, Kansas State University, Manhattan 66506.
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D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000235:genetics / Q000378:metabolism*; D004237:Diuron / Q000494:pharmacology; D017353:Gene Deletion; D015870:Gene Expression; D005798:Genes, Bacterial; D007700:Kinetics; D046911:Macromolecular Substances; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry; D045322:Photosynthetic Reaction Center Complex Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D045331:Photosystem I Protein Complex; D011190:Potassium Cyanide / Q000494:pharmacology; D015183:Restriction Mapping
D001426:Bacterial Proteins; D046911:Macromolecular Substances; D008565:Membrane Proteins; D010446:Peptide Fragments; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; C085831:photosystem I, PsaJ subunit; C079742:psaF protein, bacteria; D004237:Diuron; D011190:Potassium Cyanide