title: Targeted mutations in the psaC gene of Chlamydomonas reinhardtii: preferential reduction of FB at low temperature is not accompanied by altered electron flow from photosystem I to ferredoxin.
abstract: The terminal part of the electron pathway within the photosystem I (PSI) complex includes two [4Fe-4S] centers, FA and FB, which are coordinated by the PsaC subunit. To gain new insights into the electron transfer mechanisms through PsaC, we have generated three mutant strains of the alga Chlamydomonas reinhardtii in which two positively charged residues, K52 and R53, near the FA center have been altered in different ways. The mutations K52S/R53D and K52P/R53D lead to a strong destabilization of PSI. The third mutant K52S/R53A accumulates PSI to 30% of wild-type levels and shares the same residues between two of the cysteine ligands of FA as the PsaC homologue in the green sulfur bacterium Chlorobium limicola, in which FB has a higher redox potential than FA [Nitschke, W., Feiler, U., & Rutherford, A. W. (1990) Biochemistry 29, 3834-3842]. Low-temperature electron paramagnetic resonance (EPR) studies reveal that, in contrast to wild type, FB is preferentially photoreduced in this mutant, as was also observed for C. limicola. The preferential photoreduction of FB could be due to changes in the redox potential of FA and/or to slight structural modifications of the PsaC subunit. However, room temperature optical measurements show that stable charge separation still occurs and, surprisingly, that electron transfer from PSI to ferredoxin proceeds at normal rates in the mutant. As C. limicola, the K52S/R53A and K52S/R53D C. reinhardtii mutants are photosensitive when grown aerobically, but can grow photoautotrophically under anaerobic conditions.
authors.affiliation: Department of Molecular Biology, University of Geneva, Switzerland.
authors.forename: N
authors.initials: N
authors.lastname: Fischer
authors.forename: P
authors.initials: P
authors.lastname: Sétif
authors.forename: J D
authors.initials: JD
authors.lastname: Rochaix
mesh_terms: D000693:Anaerobiosis; D000818:Animals; D015139:Blotting, Southern; D015153:Blotting, Western; D002455:Cell Division / Q000235:genetics; D002462:Cell Membrane / Q000737:chemistry; D002696:Chlamydomonas / Q000378:metabolism*; D003001:Cloning, Molecular; D004578:Electron Spin Resonance Spectroscopy; D004579:Electron Transport / Q000235:genetics; D005288:Ferredoxins / Q000378:metabolism; D005453:Fluorescence; D017125:Genes, Protozoan / Q000235:genetics*; D007506:Iron-Sulfur Proteins / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008027:Light; D008565:Membrane Proteins; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D045322:Photosynthetic Reaction Center Complex Proteins / Q000235:genetics* / Q000378:metabolism; D045331:Photosystem I Protein Complex; D011487:Protein Conformation; D011506:Proteins / Q000235:genetics*; D013696:Temperature
chemical_list: D005288:Ferredoxins; D007506:Iron-Sulfur Proteins; D008565:Membrane Proteins; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; D011506:Proteins; C089909:photosystem I, psaB subunit