The Qo site of cytochrome b6f complexes controls the activation of the LHCII kinase.
We created a Qo pocket mutant by site-directed mutagenesis of the chloroplast petD gene in Chlamydomonas reinhardtii. We mutated the conserved PEWY sequence in the EF loop of subunit IV into PWYE. The pwye mutant did not grow in phototrophic conditions although it assembled wild-type levels of cytochrome b6f complexes. We demonstrated a complete block in electron transfer through the cytochrome b6f complex and a loss of plastoquinol binding at Qo. The accumulation of cytochrome b6f complexes lacking affinity for plastoquinol enabled us to investigate the role of plastoquinol binding at Qo in the activation of the light-harvesting complex II (LHCII) kinase during state transitions. We detected no fluorescence quenching at room temperature in state II conditions relative to that in state I. The quantum yield spectrum of photosystem I charge separation in the two state conditions displayed a trough in the absorption region of the major chlorophyll a/b proteins, demonstrating that the cells remained locked in state I. 33Pi labeling of the phosphoproteins in vivo demonstrated that the antenna proteins remained poorly phosphorylated in both state conditions. Thus, the absence of state transitions in the pwye mutant demonstrates directly that plastoquinol binding in the Qo pocket is required for LHCII kinase activation.
Zito
F
F
UPR 1261 CNRS, Institut de Biologie Physico-Chimique, 13 rue Pierre et Marie Curie, 75005 Paris, France.
Finazzi
G
G
Delosme
R
R
Nitschke
W
W
Picot
D
D
Wollman
F A
FA
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D016825:Chlamydomonas reinhardtii / Q000201:enzymology* / Q000235:genetics; D002736:Chloroplasts / Q000201:enzymology / Q000378:metabolism; D017124:Conserved Sequence / Q000235:genetics; D003573:Cytochrome b Group / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D045346:Cytochrome b6f Complex; D004578:Electron Spin Resonance Spectroscopy; D004579:Electron Transport; D004789:Enzyme Activation; D005453:Fluorescence; D007700:Kinetics; D045342:Light-Harvesting Protein Complexes; D008565:Membrane Proteins / Q000378:metabolism; D008958:Models, Molecular; D009154:Mutation; D010084:Oxidation-Reduction; D010455:Peptides / Q000378:metabolism; D010766:Phosphorylation; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism; D045331:Photosystem I Protein Complex; D010971:Plastoquinone / Q000031:analogs & derivatives / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism*; D013696:Temperature
D003573:Cytochrome b Group; D045342:Light-Harvesting Protein Complexes; D008565:Membrane Proteins; D010455:Peptides; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; C003165:plastoquinol; D045346:Cytochrome b6f Complex; D011494:Protein Kinases; C120337:light-harvesting complex II kinase; D010971:Plastoquinone