The psbH gene encodes a small protein which copurifies with photosystem 2. In the cyanobacterium Synechocystis sp. PCC 6803, psbH is located upstream of the cytochrome b6-f complex genes petC and petA. In striking contrast, in the genomes of plant chloroplasts, psbH is cotranscribed with petB and petD, encoding the other two major subunits of the cytochrome b6-f complex. We report that in Synechocystis sp. PCC 6803 monocistronic psbH and dicistronic petCA transcripts are probably initiated separately, each from DNA regions bearing some similarity to Escherichia coli sigma 70 promoters. Synechocystis sp. PCC 6803 psbH null mutants were generated by cartridge mutagenesis. Studies using a rapid screening procedure involving in situ complementation showed that the PsbH protein is not absolutely required for the assembly of a functionally active photosystem 2 complex since psbH insertion and deletion strains were able to grow photoautotrophically. The rate of photoautotrophic growth was, however, slower than the wild type, and studies of oxygen evolution, chlorophyll fluorescence, and thermoluminescence indicated that this reduction in growth rate is probably due mainly to an impairment in electron flow from QA to QB. We conclude, therefore, that the PsbH protein is not an absolute requirement for photosystem 2 activity but that it functions to optimize electron flow between the two secondary plastoquinone acceptors by interacting with the QB site on the D1 protein.
The psbK gene encodes a small protein of Photosystem II. The gene has previously been cloned and sequenced in Synechocystis sp. PCC 6803. Our new results, presented here, confirm the conclusions of Ikeuchi et al. Based on Northern hybridization and primer extension analyses, we show that psbK is transcribed as a monocistronic message in this cyanobacterium. Analysis of DNA sequence immediately upstream of the transcription start site revealed an E. coli-like-10 consensus sequence. A deletion mutant was constructed where the psbK gene was replaced by a kanamycin resistant cartridge. In situ complementation experiments, as well as Southern and Northern hybridization analyses, confirmed that the mutant strain contains a lesion in psbK. The psbK-less mutant could grow photoautotrophically as well as photoheterotrophically both in liquid culture and on agar plates. The rate of growth was slightly less compared with the wild-type as clearly observed by in situ complementation experiments. Although the mutant showed correspondingly lower rates of electron transport, thermoluminescence, oxygen flash yield and chlorophyll a fluorescence measurements did not detect any significant modification of the reactions of PS II. Moreover, the mutant was no more susceptible to excess light than the wild-type. It is, therefore, concluded that the product of the psbK gene is not crucial for PS II activity and possibly plays some other role in the metabolism of Synechocystis.
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