Inactivation of the water-oxidizing enzyme in manganese stabilizing protein-free mutant cells of the cyanobacteria Synechococcus PCC7942 and Synechocystic PCC6803 during dark incubation and conditions leading to photoactivation

Engels, Dirk H.; Lott, Alexandra; Schmid, Georg; Pistorius, Elfriede K.

doi:10.1007/bf00018265

Cited by 52 publications

(50 citation statements)

References 46 publications

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“…As shown in Fig. 5, the wild type strain was able to grow in the medium depleted of either Ca 2ϩ or Cl Ϫ with a slightly reduced rate in agreement with the results reported previously (27,28). A similar growth was observed for the strain psbU/Km.…”

Section: Cloning and Sequence Analysis Of The Psbu Genesupporting

confidence: 91%

Analysis of the psbU Gene Encoding the 12-kDa Extrinsic Protein of Photosystem II and Studies on Its Role by Deletion Mutagenesis in Synechocystis sp. PCC 6803

Shen¹,

Ikeuchi²,

Inoue³

1997

Journal of Biological Chemistry

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The gene encoding the 12-kDa extrinsic protein of photosystem II from Synechocystis sp. PCC 6803 was cloned based on N-terminal sequence of the mature protein. This gene, named psbU, encodes a polypeptide of 131 residues, the first 36 residues of which were absent in the mature protein and thus served as a transit peptide required for its transport into the thylakoid lumen. A psbU gene deletion mutant grew photoautotrophically in normal BG11 medium at almost the same rate as that of the wild type strain. This mutant, however, grew apparently slower than the wild type did upon depletion of Ca 2؉ or Cl ؊ from the growth medium. Photosystem II oxygen evolution decreased to 81% in the mutant as compared with that in the wild type, and the thermoluminescence B-and Q-bands shifted to higher temperatures accompanied by an increase in the Q-band intensity. These results indicate that the 12-kDa protein is not essential for oxygen evolution but may play a role in optimizing the ion (Ca 2؉ and Cl The oxygen-evolving system of cyanobacteria contains three extrinsic proteins, namely, a 33-kDa protein, cytochrome c-550, and a 12-kDa protein. The genes coding for the 33-kDa protein and cytochrome c-550 are psbO and psbV genes, respectively (for reviews see Refs. 1 and 2), whereas the gene for the 12-kDa protein has been tentatively named psbU (3, 4). The 33-kDa protein is commonly found in higher plant and cyanobacterial PSII, 1 and its function has been studied extensively by both in vitro biochemical approaches and in vivo mutagenesis studies.Results from these studies suggested that the 33-kDa protein plays an important role in stabilizing the tetramanganese cluster, which directly catalyzes the water-splitting reaction. Loss of this protein by biochemical removal from isolated PSII (5-7) or genetic deletion (8, 9) from algal cells leads to a significant loss of the oxygen-evolving activity and in some conditions loss of manganese atoms from the tetramanganese cluster. The other two proteins, cytochrome c-550 and the 12-kDa protein, however, are present only in algal-type PSII but absent in higher plant PSII. Our previous in vitro biochemical (10) and in vivo genetic studies (11) have indicated that cytochrome c-550 is required for maintaining both the oxygen evolution and PSII stability in vivo. This cytochrome can bind to PSII essentially independent of the other extrinsic proteins (10). In accordance with this, a double deletion mutant of Synechocystis sp. PCC 6803 lacking both the 33-kDa protein and cytochrome c-550 showed a complete loss of photoautotrophic growth, which is caused by deactivation of oxygen evolution and destabilization of PSII in vivo (12). In contrast, both the single deletion mutant of the 33-kDa protein (9) or cytochrome c-550 (11) were able to grow autotrophically, albeit with reduced rates. These studies suggested that cytochrome c-550 binds to and functions in cyanobacterial PSII independent of the 33-kDa protein in maintaining the oxygen-evolving activity and PSII stability (12). The 12-kDa pro...

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Section: Cloning and Sequence Analysis Of The Psbu Genesupporting

confidence: 91%

Analysis of the psbU Gene Encoding the 12-kDa Extrinsic Protein of Photosystem II and Studies on Its Role by Deletion Mutagenesis in Synechocystis sp. PCC 6803

Shen¹,

Ikeuchi²,

Inoue³

1997

Journal of Biological Chemistry

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“…In contrast, the PsbY-free Synechocystis PCC 6803 mutant produced almost as much 18 O 2 from H 2 18 O 2 as 16 O 2 from H 2 16 O. For comparison we also present the results of an experiment with the previously constructed PsbO-free Synechocystis PCC 6803 mutant (Engels et al 1994), which also shows significantly higher 18 O 2 evolution from H 2 18 O 2 than its parental wild-type strain (Fig. 5).…”

Section: Photosynthetic Oxygen Evolutionmentioning

confidence: 77%

“…strain PCC 6803 (hereafter referred to as Synechocystis PCC 6803 WT) and its PsbY-free mutant derivative were obtained from the laboratory of Prof. Dr. S. V. Shestakov (Department of Genetics, Biology Division, Moscow State University, Moscow, Russia). The previously constructed PsbO-free mutant (Engels et al 1994;Stephan et al 2000) was derived from a wild-type Synechocystis PCC 6803 strain obtained from the Pasteur Culture Collection of Cyanobacterial Strains (Paris, France), and both of these strains were also used in this present work.…”

Section: Cyanobacterial Strainsmentioning

confidence: 99%

“…The PsbY-free Synechocystis PCC 6803 mutant was grown under the same conditions as PCC 6803 WT, except that kanamycin sulfate (40 mg/l) was included in the medium. The Synechocystis PCC 6803 strain from which the PsbOfree mutant was derived (Engels et al 1994) was also grown as described above. The growth medium for the PsbO-free mutant contained kanamycin sulfate at 7.5 mg/l (Stephan et al 2000).…”

Section: Growth Conditionsmentioning

confidence: 99%

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On the functional significance of the polypeptide PsbY for photosynthetic water oxidation in the cyanobacterium Synechocystis sp. strain PCC 6803

et al. 2004

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Recent investigations have revealed that the cyanobacterial photosystem II complex contains more than 26 polypeptides. The functions of most of the low-molecular-mass polypeptides, including PsbY, have remained elusive. Here we present a comparative characterization of the wild-type Synechocystis sp. strain PCC 6803 and a PsbY-free mutant derived from it. The results show that growth of the PsbY-free mutant was comparable to that of the wild-type when cells were cultivated in complete BG11 medium or under initial manganese or chloride limitation, and when illuminated at 20 or 200 microE m(-2) s(-1). However, while growth rates of both the wild-type and the PsbY-free mutant were reduced when cells were cultivated in BG11 medium in the absence of calcium, the reduction was significantly greater in the case of the PsbY-free mutant. This differential effect on growth of the mutant relative to the wild-type in CaCl(2) deficient medium was detected when the cells were illuminated with high-intensity light (200 microE m(-2) s(-1)) but not when light levels were lower (20 microE m(-2) s(-1)). The differential effect on growth was associated with lower O(2) evolving activity in the mutant compared to wild-type cells. The mutant was also found to be more sensitive to photoinhibition, and showed an altered pattern of fluorescence emission at 77 K. In addition, mass spectrometric analysis revealed that PsbY-free cells cultivated in CaCl(2) sufficient medium (in which no growth reduction was observed) had a significantly higher O(2) evolution from hydrogen peroxide and a lower O(2) evolution from water under flash light illumination than wild-type cells. These results imply that photosystem II is slightly impaired in the PsbY-free mutant, and that the mutant is less capable of coping with low levels of Ca(2+) than the wild-type.

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“…Under these conditions the O 2 evolving activity of the mutant was almost as high as that of WT (maximally about 5% lower than that of WT) (not shown). For comparison we measured the O 2 evolving rate of a previously constructed PsbOfree Synechocystis PCC 6803 mutant (Engels et al, 1994) which has been shown to be extremely light sensitive (Philbrick et al, 1991). Therefore, the growth deficiency of Hik14 is not due to a higher sensitivity towards photoinhibition as the primary (Grossman et al, 2001;Hsiao et al, 2004;Murata and Suzuki, 2006).…”

Section: ð1mentioning

confidence: 99%

Physiological and Molecular Characterization of a Synechocystis sp. PCC 6803 Mutant Lacking Histidine Kinase Slr1759 and Response Regulator Slr1760

Nodop

Suzuki

Barsch

et al. 2006

Zeitschrift Für Naturforschung C

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The hybrid sensory histidine kinase Slr1759 of the cyanobacterium Synechocystis sp. strain PCC 6803 contains multiple sensory domains and a multi-step phosphorelay system. Immuno blot analysis provided evidence that the histidine kinase Slr1759 is associated with the cytoplasmic membrane. The gene slr1759 is part of an operon together with slr1760, encoding a response regulator. A comparative investigation was performed on Synechocystis sp. strain PCC 6803 wild type (WT) and an insertionally inactivated slr1759-mutant (Hik14) which also lacks the transcript for the response regulator Slr1760. The mutant Hik14 grew significantly slower than WT in the early growth phase, when both were inoculated with a low cell density into BG11 medium without additional buffer and when aerated with air enriched with 2% CO 2 . Since the aeration with CO 2 -enriched air results in a decrease of the pH value in the medium, the growth experiments indicated that Hik14 is not able to adjust its metabolic activities as rapidly as WT to compensate for a larger decrease of the pH value in the medium. No significant differences in growth between Hik14 and WT were observed when cells were inoculated with a higher cell density in BG11 medium or when the BG11 medium contained 50 mm Epps-NaOH, pH 7.5, to prevent the pH drop. This Hik14 phenotype has so far only been seen under the above defined growth condition. Results of photosynthetic activity measurements as well as Northern blot-, immuno blot-, and metabolite analyses suggest that the two-component system Slr1759/Slr1760 has a function in the coordination of several metabolic activities which is in good agreement with the complex domain structure of Slr1759. The direct targets of this two-component system have so far not been identified.

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Inactivation of the water-oxidizing enzyme in manganese stabilizing protein-free mutant cells of the cyanobacteria Synechococcus PCC7942 and Synechocystic PCC6803 during dark incubation and conditions leading to photoactivation

Cited by 52 publications

References 46 publications

Analysis of the psbU Gene Encoding the 12-kDa Extrinsic Protein of Photosystem II and Studies on Its Role by Deletion Mutagenesis in Synechocystis sp. PCC 6803

Analysis of the psbU Gene Encoding the 12-kDa Extrinsic Protein of Photosystem II and Studies on Its Role by Deletion Mutagenesis in Synechocystis sp. PCC 6803

On the functional significance of the polypeptide PsbY for photosynthetic water oxidation in the cyanobacterium Synechocystis sp. strain PCC 6803

Physiological and Molecular Characterization of a Synechocystis sp. PCC 6803 Mutant Lacking Histidine Kinase Slr1759 and Response Regulator Slr1760

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