Knock-out of the chloroplast-encoded PSI-J subunit of photosystem I in Nicotiana tabacum.
The plastid-encoded psaJ gene encodes a hydrophobic low-molecular-mass subunit of photosystem I (PSI) containing one transmembrane helix. Homoplastomic transformants with an inactivated psaJ gene were devoid of PSI-J protein. The mutant plants were slightly smaller and paler than wild-type because of a 13% reduction in chlorophyll content per leaf area caused by an approximately 20% reduction in PSI. The amount of the peripheral antenna proteins, Lhca2 and Lhca3, was decreased to the same level as the core subunits, but Lhca1 and Lhca4 were present in relative excess. The functional size of the PSI antenna was not affected, suggesting that PSI-J is not involved in binding of light-harvesting complex I. The specific PSI activity, measured as NADP(+) photoreduction in vitro, revealed a 55% reduction in electron transport through PSI in the mutant. No significant difference in the second-order rate constant for electron transfer from reduced plastocyanin to oxidized P700 was observed in the absence of PSI-J. Instead, a large fraction of PSI was found to be inactive. Immunoblotting analysis revealed a secondary loss of the luminal PSI-N subunit in PSI particles devoid of PSI-J. Presumably PSI-J affects the conformation of PSI-F, which in turn affects the binding of PSI-N. This together renders a fraction of the PSI particles inactive. Thus, PSI-J is an important subunit that, together with PSI-F and PSI-N, is required for formation of the plastocyanin-binding domain of PSI. PSI-J is furthermore important for stability or assembly of the PSI complex.
Hansson
Andreas
A
Plant Biochemistry Laboratory, Department of Plant Biology, Faculty of Life Sciences, University of Copenhagen, Frederiksberg, Denmark.
Amann
Katrin
K
Zygadlo
Agnieszka
A
Meurer
Jörg
J
Scheller
Henrik V
HV
Jensen
Poul E
PE
D002734:Chlorophyll / Q000032:analysis / Q000378:metabolism; D000077194:Chlorophyll A; D025222:Chloroplast Proton-Translocating ATPases / Q000032:analysis; D002736:Chloroplasts / Q000235:genetics / Q000378:metabolism*; D004579:Electron Transport; D020868:Gene Silencing; D007700:Kinetics; D045342:Light-Harvesting Protein Complexes / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction; D010641:Phenotype; D045331:Photosystem I Protein Complex / Q000235:genetics / Q000378:metabolism*; D045332:Photosystem II Protein Complex / Q000235:genetics / Q000378:metabolism; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D010970:Plastocyanin / Q000378:metabolism; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D020524:Thylakoids / Q000737:chemistry / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*
D045342:Light-Harvesting Protein Complexes; D045331:Photosystem I Protein Complex; D045332:Photosystem II Protein Complex; D010940:Plant Proteins; D021122:Protein Subunits; C085831:photosystem I, PsaJ subunit; D002734:Chlorophyll; D009249:NADP; C001785:chlorophyll P 700; C037184:chlorophyll b; D010970:Plastocyanin; D025222:Chloroplast Proton-Translocating ATPases; D000077194:Chlorophyll A