Purification and Characterisation of the Carboxyl-Terminal Processing Protease of the D1 Protein of Photosystem II from Scenedesmus Obliquus and Pisum Sativum

Packer, Jeremy C.L.; Taylor, Mark A.; Gerrish, Christopher; Bowyer, John R.

doi:10.1007/978-94-009-0511-5_603

Cited by 2 publications

(2 citation statements)

References 8 publications

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“…The results presented here would suggest that the protease present in Synechocystis 6803 can recognize and cleave a higher plant D1 precursor molecule, even though there is a high degree of divergence between cyanobacteria and green plants in the primary structure of the carboxyl-terminal region of D1. In support of this idea of broad substrate specificity is the recent work demonstrating that the purified processing enzyme from both spinach (Inagaki et al, 1990) and pea (Packer et al, 1990) can process the D1 polypeptide found in the LFI mutant of the green alga S.…”

Section: Processing Of the D1 Polypeptidementioning

confidence: 89%

Expression of a higher plant psbA gene in Synechocystis 6803 yields a functional hybrid photosystem II reaction center complex.

1991

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The psbA gene codes for the D1 polypeptide of the photosystem I 1 reaction center complex and is found in a11 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 posttranslationally modified at the carboxyl terminus, probably through processing. A detailed biophysical analysis of the chimeric photosystem I 1 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 S p ) 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 I 1 reaction center.

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Section: Processing Of the D1 Polypeptidementioning

confidence: 89%

Expression of a higher plant psbA gene in Synechocystis 6803 yields a functional hybrid photosystem II reaction center complex.

1991

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“…In support of this idea of broad substrate specificity is the recent work demonstrating that the puri? fied processing enzyme from both spinach (Inagaki et al, 1990) and pea (Packer et al, 1990) can process the D1 polypeptide found in the LF1 mutant of the green alga S. obliquus, despite a conservation of only 5 out of 9 residues (Todd et al, 1990) after the proposed cleavage site on the carboxyl side of alanine 344. Presumably, sequences within the mature portion of the D1 polypeptide are more important for recognition by the protease.…”

Section: Processing Of the D1 Polypeptidementioning

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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Purification and Characterisation of the Carboxyl-Terminal Processing Protease of the D1 Protein of Photosystem II from Scenedesmus Obliquus and Pisum Sativum

Cited by 2 publications

References 8 publications

Expression of a higher plant psbA gene in Synechocystis 6803 yields a functional hybrid photosystem II reaction center complex.

Expression of a higher plant psbA gene in Synechocystis 6803 yields a functional hybrid photosystem II reaction center complex.

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

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