A large fraction of PsaF is nonfunctional in photosystem I complexes lacking the PsaJ subunit.
PsaJ is a small hydrophobic subunit of the photosystem I complex (PSI) whose function is not yet fully understood. Here we describe mutants of the green alga Chlamydomonas reinhardtii, in which the psaJ chloroplast gene has been inactivated either in a wild-type or in a PsaF-deficient nuclear background. Cells lacking one or both subunits grow photoautotrophically and contain normal levels of PSI. Flash-absorption spectroscopy performed with isolated PSI particles isolated from the PsaJ-deficient strain indicates that only 30% of the PSI complexes oxidize plastocyanin (Pc) or cytochrome c6 (Cyt c6) with kinetics identical to wild type, whereas the remaining 70% follow slow kinetics similar to those observed with PsaF-deficient PSI complexes. This feature is not due to partial loss of PsaF, as the PsaJ-less PSI complex contains normal levels of the PsaF subunit. The N-terminal domain of PsaF can be cross-linked to Pc and Cyt c6 indicating that in the absence of PsaJ, this domain is exposed in the lumenal space. Therefore, the decreased amount of functional PsaF revealed by the electron-transfer measurements is best explained by a displacement of the N-terminal domain of PsaF which is known to provide the docking site for Pc and Cyt c6. We propose that one function of PsaJ is to maintain PsaF in a proper orientation which allows fast electron transfer from soluble donor proteins to P700(+).
Fischer
N
N
Department of Molecular Biology, University of Geneva, Switzerland.
Boudreau
E
E
Hippler
M
M
Drepper
F
F
Haehnel
W
W
Rochaix
J D
JD
D000595:Amino Acid Sequence; D000818:Animals; D016825:Chlamydomonas reinhardtii / Q000235:genetics / Q000378:metabolism; D002736:Chloroplasts / Q000235:genetics; D003432:Cross-Linking Reagents / Q000378:metabolism; D003580:Cytochromes / Q000378:metabolism; D045348:Cytochromes f; D004579:Electron Transport; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D007425:Intracellular Membranes / Q000378:metabolism; D045342:Light-Harvesting Protein Complexes; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D045322:Photosynthetic Reaction Center Complex Proteins / Q000235:genetics* / Q000378:metabolism*; D045331:Photosystem I Protein Complex; D010940:Plant Proteins / Q000378:metabolism; D010970:Plastocyanin / Q000378:metabolism; D015800:Protozoan Proteins
D003432:Cross-Linking Reagents; D003580:Cytochromes; D045342:Light-Harvesting Protein Complexes; D008565:Membrane Proteins; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; D010940:Plant Proteins; D015800:Protozoan Proteins; C085831:photosystem I, PsaJ subunit; C489801:psaF protein, Chlamydomonas reinhardtii; D010970:Plastocyanin; D045348:Cytochromes f