The chloroplast-encoded alpha subunit of cytochrome b-559 is required for assembly of the photosystem two complex in both the light and the dark in Chlamydomonas reinhardtii.
The role of cytochrome b-559 in the photosystem two (PSII) complex has been investigated through the construction of a psbE null mutant by transformation of the chloroplast genome of the green alga Chlamydomonas reinhardtii. No PSII activity could be detected in this mutant either in oxygen evolution assays or by analysis of variable chlorophyll fluorescence. Immunoblotting experiments showed that the absence of PSII activity in the mutant was due to the loss of the PSII complex in both light-grown and dark-grown cultures. In contrast, the photosystem one reaction center polypeptide, PsaA, was present at wild-type levels in the mutant. RNA gel blot assays confirmed that the transcript levels for the psbA, psbD, and psbF genes were unaffected by disruption of the psbE gene, suggesting a post-transcriptional effect on their expression. Pulse-labeling experiments showed that either synthesis of PSII subunits was impaired in the psbE null mutant or there was extremely rapid degradation of newly synthesized subunits. Interestingly, the PsbE and PsbF subunits accumulated to wild-type levels in a psbA deletion mutant of C. reinhardtii, FuD7, which fails to synthesize D1 and assemble PSII. Our results provide evidence for a role for cytochrome b-559 in the early steps of assembly of the PSII complex, possibly as a redox-controlled nucleation factor that determines the level of PSII within the thylakoid membrane.
Morais
F
F
Wolfson Laboratories, Department of Biochemistry, Imperial College of Science, Technology and Medicine, London, SW7 2AY, United Kingdom.
Barber
J
J
Nixon
P J
PJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016825:Chlamydomonas reinhardtii / Q000201:enzymology*; D002736:Chloroplasts / Q000201:enzymology*; D003573:Cytochrome b Group / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers; D003624:Darkness; D015151:Immunoblotting; D008027:Light; D045342:Light-Harvesting Protein Complexes; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism*; D045332:Photosystem II Protein Complex; D012333:RNA, Messenger / Q000235:genetics
D003573:Cytochrome b Group; D017931:DNA Primers; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D012333:RNA, Messenger; C019313:cytochrome b559