Insertion of a Chloroplast psbA Gene Into the Chromosome of the Cyanobacterium Synechocystis 6803

Reiss, Thomas; Jansson, Christer; McIntosh, Lee

doi:10.1007/978-94-009-0511-5_578

Cited by 4 publications

(3 citation statements)

References 7 publications

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“…The three following Synechocystis 6803 DNA probes were used: an internal fragment of the psbA-2 gene [ 5,11 ], a fragment containing 5' coding and non-coding regions of the rbcL gene [ 11,17] and an internal fragment of the psbD-2…”

Section: Rna Isolation Electrophoresis and Northern Blot Analysesmentioning

confidence: 99%

Photosynthetic electron transport controls degradation but not production of psbA transcripts in the cyanobacterium Synechocystis 6803

Mohamed

Jansson

1991

Plant Mol Biol

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Accumulation and stability of psbA and rbcL-S transcripts in Synechocystis 6803 was followed in the presence and absence of the photosynthesis inhibitors DCMU and methylviologen. Our results demonstrate that both transcript production and transcript stability are important regulatory elements for psbA gene expression in Synechocystis 6803. The production of psbA transcripts was stimulated by light in a process that operated independently of the photosynthetic electron transport. However, stability of the psbA transcript increased in the dark and was controlled by photosynthetic electron transport. The psbA transcript was remarkably stable in the dark, with a half-life of approximately 7 hours. By contrast, the regulatory pattern for the rbcL-S genes was quite different. The light-stimulated production of rbcL-S transcripts was dependent on an intact photosynthetic electron transport, and rbcL-S transcript stability was higher under illuminated conditions than in darkness.

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Section: Rna Isolation Electrophoresis and Northern Blot Analysesmentioning

confidence: 99%

Photosynthetic electron transport controls degradation but not production of psbA transcripts in the cyanobacterium Synechocystis 6803

Mohamed

Jansson

1991

Plant Mol Biol

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“…Reiss et al (1990) have reported recently the incorporation of the psbA gene from A. hybridus into the chromosome of a strain of Synechocystis 6803 in which the psbA gene family had been inactivated by insertional mutagenesis. The resulting strain grew photoautotrophically.…”

Section: Other Attempts At Gene Substitutionmentioning

confidence: 99%

Expression of a Higher Plant psbA Gene in Synechocystis 6803 Yields a Functional Hybrid Photosystem II Reaction Center Complex

Nixon¹,

Rögner²,

Diner³

1991

The Plant Cell

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The psbA gene codes for the D1 polypeptide of the photosystem II reaction center complex and is found in all photosynthetic organisms that carry out oxygenic photosynthesis. Here we describe the construction and characterization of a strain of the cyanobacterium Synechocystis sp PCC 6803 in which the three endogenous psbA genes are replaced by a single psbA gene from the chloroplast genome of the higher plant Poa annua. The resulting chimeric strain, KWPAS, grows photoautotrophically with a doubling time of 26 hours compared with 20 hours for wild-type Synechocystis 6803. The mutant oxidizes water to oxygen at light-saturated rates comparable with wild type, despite differences in 15% of the primary structure of D1 between these species. RNA gel blot analysis indicates the presence in KWPAS of a psbA transcript of approximately 1.25 kilobases, consistent with the chloroplast promoter also acting as a promoter in Synechocystis. By using antibodies specific for the carboxyl-terminal extension of the D1 polypeptide of higher plants, we showed that the D1 polypeptide synthesized by KWPAS is post-translationally modified at the carboxyl terminus, probably through processing. A detailed biophysical analysis of the chimeric photosystem II complex indicated that the rates of forward electron transfer are similar to wild type. The rates of charge recombination between the donor and acceptor sides of the reaction center are, however, accelerated by as much as a factor of nine (QA- to S2) and are the most likely explanation for the lower rate of photoautotrophic growth in the mutant. We conclude that the psbA gene from a higher plant can be expressed in cyanobacteria and its product processed and assembled into a functional chimeric photosystem II reaction center.

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“…We have previously demonstrated that this insertion is dispensable for photosynthetic growth of Synechcocystis 6803 by replacing the cyanobacterial psbA gene with that of Amaranthus hybridus (Reiss et al 1990). The resultant mutant produced a functional, chimaeric PS II complex.…”

Section: Construction and Genetic Characterization Of Mutantsmentioning

confidence: 99%

Constructed deletions in lumen-exposed regions of the D1 protein in the cyanobacterium Synechocystis 6803: Effects on D1 insertion and accumulation in the thylakoid membrane, and on Photosystem II assembly

et al. 1996

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Modified forms of the D1 protein with deletions in lumen-exposed regions, were constructed in the cyanobacterium Synechocystis 6803 using site-directed mutagenesis. Integration and stability of the mutated D1 proteins in the thylakoid membrane were studied by immunoblot and pulse-chase analyses. It was found that in Δ(N325-E333), the D1 protein with a deletion in the C-terminal tail, could insert in the thylakoids to normal amounts but its stability in the membrane was dramatically reduced. Insertion of D1 in Δ(V58-D61) or Δ(D103-G109);G110R, with deletions in the A-B loop, was severely obstructed, For Δ(P350-T354), with a deletion in the processed region of the C-terminus of D1, no phenotypic effects were observed. The effects of failed D1 insertion or accumulation on Photosystem II assembly was monitored by immunoblot analysis. The conclusions from these experiments are that the extrinsic 33 kDa protein, CP43, and the β subunit of cytochrome b559 accumulate in the thylakoid membrane independently of the D1 protein, and that accumulation of the D2 protein and CP47 requires insertion but not necessarily accumulation of the D1 protein.

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Insertion of a Chloroplast psbA Gene Into the Chromosome of the Cyanobacterium Synechocystis 6803

Cited by 4 publications

References 7 publications

Photosynthetic electron transport controls degradation but not production of psbA transcripts in the cyanobacterium Synechocystis 6803

Photosynthetic electron transport controls degradation but not production of psbA transcripts in the cyanobacterium Synechocystis 6803

Expression of a Higher Plant psbA Gene in Synechocystis 6803 Yields a Functional Hybrid Photosystem II Reaction Center Complex

Constructed deletions in lumen-exposed regions of the D1 protein in the cyanobacterium Synechocystis 6803: Effects on D1 insertion and accumulation in the thylakoid membrane, and on Photosystem II assembly

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