Publications
173 results found
Andronis C, Durrant JR, Diner BA, et al., 1995, Construction and initial characterisation of a D2-Leu205Tyr mutant of Chlamydomonas reinhardtii, Xth International Photosynthesis Congress, Publisher: KLUWER ACADEMIC PUBL, Pages: 839-842
KRUIP J, NIXON PJ, OSIEWACZ HD, et al., 1994, NUCLEOTIDE-SEQUENCE OF THE PETB GENE ENCODING CYTOCHROME B(6) FROM THE MESOPHILIC CYANOBACTERIUM SYNECHOCYSTIS PCC-6803 - IMPLICATIONS FOR EVOLUTION AND FUNCTION, BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS, Vol: 1188, Pages: 443-446, ISSN: 0005-2728
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- Citations: 3
ALIZADEH S, NECHUSHTAI R, BARBER J, et al., 1994, NUCLEOTIDE-SEQUENCE OF THE PSBE, PSBF AND TRNM GENES FROM THE CHLOROPLAST GENOME OF CHLAMYDOMONAS-REINHARDTII, BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS, Vol: 1188, Pages: 439-442, ISSN: 0005-2728
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- Citations: 9
GIORGI LB, DURRANT JR, ALIZADEH S, et al., 1994, COMPARISON OF PRIMARY ELECTRON-TRANSFER IN PHOTOSYSTEM-II REACTION CENTERS ISOLATED FROM THE HIGHER-PLANT PISUM-SATIVUM AND THE GREEN-ALGA CHLAMYDOMONAS-REINHARDTII, BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS, Vol: 1186, Pages: 247-251, ISSN: 0005-2728
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- Citations: 9
DALLACHIESA M, MAYERS SR, MASKELL DJ, et al., 1994, AN AROA HOMOLOG FROM SYNECHOCYSTIS SP PCC6803, GENE, Vol: 144, Pages: 145-146, ISSN: 0378-1119
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- Citations: 2
NIXON PJ, DINER BA, 1994, ANALYSIS OF WATER-OXIDATION MUTANTS CONSTRUCTED IN THE CYANOBACTERIUM SYNECHOCYSTIS SP PCC-6803, BIOCHEMICAL SOCIETY TRANSACTIONS, Vol: 22, Pages: 338-343, ISSN: 0300-5127
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- Citations: 63
Nixon PJ, Trost JT, Diner BA, 1992, Role of the carboxy terminus of polypeptide D1 in the assembly of a functional water-oxidizing manganese cluster in photosystem II of the cyanobacterium Synechocystis sp. PCC 6803: assembly requires a free carboxyl group at C-terminal position 344., Biochemistry, Vol: 31, Pages: 10859-10871, ISSN: 0006-2960
The D1 polypeptide of the photosystem II (PSII) reaction center is synthesized as a precursor polypeptide which is posttranslationally processed at the carboxy terminus. It has been shown in spinach that such processing removes nine amino acids, leaving Ala344 as the C-terminal residue [Takahashi, M., Shiraishi, T., & Asada, K. (1988) FEBS Lett. 240, 6-8; Takahashi, Y., Nakane, H., Kojima, H., & Satoh, K. (1990) Plant Cell Physiol. 31, 273-280]. We show here that processing on the carboxy side of Ala344 also occurs in the cyanobacterium Synechocystis 6803, resulting in the removal of 16 amino acids. By constructing a deletion strain of Synechocystis 6803 that lacks the three copies of the psbA gene encoding D1, we have developed a system for generating psbA mutants. Using this system, we have constructed mutants of Synechocystis 6803 that are modified in the region of the C-terminus of the D1 polypeptide. Characterization of these mutants has revealed that (1) processing of the D1 polypeptide is blocked when the residue after the cleavage site is changed from serine to proline (mutant Ser345Pro) with the result that the manganese cluster is unable to assemble correctly; (2) the C-terminal extension of 16 amino acid residues can be deleted with little consequence either for insertion of D1 into the thylakoid membrane or for assembly of D1 into a fully active PSII complex; (3) removal of only one more residue (mutant Ala344stop) results in a loss of assembly of the manganese cluster; and (4) the ability of detergent-solubilized PSII core complexes (lacking the manganese cluster) to bind and oxidize exogenous Mn2+ by the secondary donor, Z+, is largely unaffected in the processing mutants (the Ser345Pro mutant of Synechocystis 6803 and the LF-1 mutant of Scenedesmus obliquus) and the truncation mutant Ala344stop. Our results are consistent with a role for processing in regulating the assembly of the photosynthetic manganese cluster and a role for the free carbo
Nixon PJ, Diner BA, 1992, Aspartate 170 of the photosystem II reaction center polypeptide D1 is involved in the assembly of the oxygen-evolving manganese cluster., Biochemistry, Vol: 31, Pages: 942-948, ISSN: 0006-2960
Eleven site-directed mutations were constructed at aspartate 170 of the D1 polypeptide of the photosystem II (PSII) reaction center of the cyanobacterium Synechocystis sp. PCC 6803. The light-saturated rates of O2 evolution (VO2) measured in whole cells range from close to that of wild-type for Asp170Glu to zero for Asp170Ser and Ala. Those mutant strains that are best able to evolve O2 are also those that show the lowest Km in PSII core complexes for the oxidation of Mn2+ by oxidized Tyr161, the normal oxidant of the Mn cluster responsible for O2 evolution. To a first approximation, the lower the pKa of the residue at position 170, the higher the VO2 and the lower the Km. D1-Asp170 appears to participate in the early steps associated with the assembly of the Mn cluster. It is also the first reported example of an amino acid residue critical to the function and assembly of the oxygen-evolving complex.
Diner B A, Nixon P J, 1992, The rate of reduction of oxidized redox-active tyrosine, Z+, by exogenous Mn2+ is slowed in a site-directed mutant, at aspartate 170 of polypeptide D1 of photosystem II, inactive for photosynthetic oxygen evolution, Biochim. Biophys. Acta, Vol: 1101, Pages: 134-138
P J Nixon, D A Chisholm, B A Diner, 1992, Isolation and functional analysis of random and site-directed mutants of photosystem II, Plant Protein Engineering, Editors: Shewry, Gutteridge, Cambridge, UK, Publisher: Cambridge University Press, Pages: 93-141
Nixon PJ, Rögner M, Diner BA, 1991, Expression of a higher plant psbA gene in Synechocystis 6803 yields a functional hybrid photosystem II reaction center complex., Plant Cell, Vol: 3, Pages: 383-395, ISSN: 1040-4651
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.
Diner B A, Nixon P J, Farchaus J W, 1991, Site-directed mutagenesis of photosynthetic reaction centers, Curr. Opin. Struct. Biol., Vol: 1, Pages: 546-554
Rögner M, Nixon PJ, Diner BA, 1990, Purification and characterization of photosystem I and photosystem II core complexes from wild-type and phycocyanin-deficient strains of the cyanobacterium Synechocystis PCC 6803., J Biol Chem, Vol: 265, Pages: 6189-6196, ISSN: 0021-9258
Highly photoactive Photosystem I (PS I) and Photosystem II (PS II) core complexes have been isolated from the cyanobacterium Synechocystis Pasteur Culture Collection (PCC) 6803 and a phycocyanin-deficient mutant, enriched in PS II. Cell breakage using glass beads was followed by sucrose density gradient centrifugation and two high-performance liquid chromatography steps involving anion-exchange and hydroxyapatite. The PS I core complex has an apparent molecular mass of 300 +/- 20 kDa (including a detergent shell of about 50 kDa) and contains subunits of approximately 60, approximately 60, 18.5, 18.5, 16, 15, 10.5, 9.5, and 6.5 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblots; its antenna size is 75 +/- 5 chlorophyll/P-700. The PS II core complex has an apparent molecular mass of 310 +/- 20 kDa (including the detergent shell); subunits of 43, 37, 33, 29, and 10-11 kDa were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. The antenna size of the average PS II complex is 45 +/- 5 chlorophyll/primary quinone electron acceptor (QA). This preparation procedure also yields, as a byproduct, a highly purified cytochrome b6f complex. This complex contains four subunits of 38, 24, 19, and 15 kDa and b- and c-type cytochromes in a ratio of 2:1. Its apparent molecular mass of 180 +/- 20 kDa (including the detergent shell) is consistent with a monomeric complex.
DINER BA, NIXON PJ, METZ JG, et al., 1990, PHOTOSYSTEM-II ELECTRON-DONOR AND ACCEPTOR-SIDE FUNCTION PROBED BY SITE-DIRECTED MUTANTS, BIOPHYSICAL JOURNAL, Vol: 57, Pages: A405-A405, ISSN: 0006-3495
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- Citations: 7
Metz J, Nixon P, Diner B, 1990, Nucleotide sequence of the psbA3 gene from the cyanobacterium Synechocystis PCC 6803, Nucl. Acids Res., Vol: 18
Nixon PJ, Gounaris K, Coomber SA, et al., 1989, psbG is not a photosystem two gene but may be an ndh gene., J Biol Chem, Vol: 264, Pages: 14129-14135, ISSN: 0021-9258
A gene of the chloroplast genome has been designated the psbG gene on the basis that in maize the gene product is a 24-kDa polypeptide of photosystem two (PS2) (Steinmetz, A. A., Castroviejo, M., Sayre, R. T., and Bogorad, L. (1986) J. Biol. Chem. 261, 2485-2488). We have located and sequenced the equivalent gene in wheat (Triticum aestivum) and have raised specific antibodies to the gene product following its expression in Escherichia coli as a beta-galactosidase fusion protein. Using these antibodies, we have investigated the location of the gene product in various thylakoid membrane fractions of pea (Pisum sativum). The gene product of apparent molecular mass 27-28 kDa is severely depleted in PS2-enriched membrane preparations and its distribution between stromal and granal regions of the membrane is distinct to that of the psbC gene product which is known to be a core polypeptide of PS2. We therefore conclude that psbG does not code for a component of PS2 but instead suggest that it is present in a novel protein complex of the thylakoid membrane. On the basis of 1) the conserved overlap between psbG and ndhC, a chloroplast gene which shows significant homology to a mitochondrial gene that codes for a subunit of the NADH-ubiquinone oxidoreductase of mitochondria, and 2) sequence similarity between the psbG gene product and the ndh gene product of E. coli, which codes for a respiratory NADH dehydrogenase, we propose that this ill-defined complex functions as a NADH or NADPH-plastoquinone oxidoreductase.
Metz JG, Nixon PJ, Rögner M, et al., 1989, Directed alteration of the D1 polypeptide of photosystem II: evidence that tyrosine-161 is the redox component, Z, connecting the oxygen-evolving complex to the primary electron donor, P680., Biochemistry, Vol: 28, Pages: 6960-6969, ISSN: 0006-2960
In photosystem II, electrons are sequentially extracted from water at a site containing Mn atoms and transferred through an intermediate carrier (Z) to the photooxidized reaction-center chlorophyll (P680+). Two polypeptides, D1 and D2, coordinate the primary photoreactants of the reaction center. Recently Debus et al. [Debus, R.J., Barry, B.A., Babcock, G.T., & McIntosh, L. (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 427-430], have suggested that Z is a tyrosine residue located at position 161 of the D1 protein. To test this proposal, we have engineered a strain of the cyanobacterium Synechocystis PCC 6803 to produce a D1 polypeptide in which Tyr-161 has been replaced by phenylalanine. Wild-type Synechocystis PCC 6803 contains three nonidentical copies of the psbA gene which encode the D1 polypeptide. In the mutant strain, two copies were deleted by replacement with antibiotic-resistance genes, and site-directed mutations were constructed in a cloned portion of the remaining gene (psbA-3), carrying a third antibiotic-resistance gene downstream. Transformants were selected for antibiotic resistance and then screened for a photoautotrophy-minus phenotype. The mutant genotype was verified by complementation tests and by amplification and sequencing of genomic DNA. Cells of the mutant cannot evolve oxygen and, unlike the wild type, are unable to stabilize, with high efficiency, the charge-separated state in the presence of hydroxylamine and DCMU [3-(3,4-dichlorophenyl)-1,1-dimethylurea]. Analyses by optical and EPR spectroscopy of reaction centers purified from this mutant indicate that Z can no longer be photooxidized and, instead, a chlorophyll cation radical, Chl+, is produced in the light. In the wild type, charge recombination between Z+ and the reduced primary quinone electron acceptor QA- occurs with a t1/2 of 80 ms. In the mutant, charge recombination between Chl+ and QA- occurs with a t1/2 of 1 ms. From these observations, we conclude that Z is indeed Tyr-161
TAYLOR MA, NIXON PJ, TODD CM, et al., 1988, CHARACTERIZATION OF THE D1 PROTEIN IN A PHOTOSYSTEM-II MUTANT (LF-1) OF SCENEDESMUS-OBLIQUUS BLOCKED ON THE OXIDIZING SIDE - EVIDENCE SUPPORTING NON-PROCESSING OF D1 AS THE CAUSE OF THE LESION, FEBS LETTERS, Vol: 235, Pages: 109-116, ISSN: 0014-5793
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- Citations: 27
MARDER JB, CHAPMAN DJ, TELFER A, et al., 1987, IDENTIFICATION OF PSBA AND PSBD GENE-PRODUCTS, D1 AND D2, AS REACTION CENTER PROTEINS OF PHOTOSYSTEM-2, PLANT MOLECULAR BIOLOGY, Vol: 9, Pages: 325-333, ISSN: 0167-4412
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- Citations: 94
NIXON PJ, DYER TA, BARBER J, et al., 1986, IMMUNOLOGICAL EVIDENCE FOR THE PRESENCE OF THE D 1-PROTEIN AND D 2-PROTEIN IN PS-II CORES OF HIGHER-PLANTS, FEBS LETTERS, Vol: 209, Pages: 83-86, ISSN: 0014-5793
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- Citations: 39
Zhang P, Sicora C, Vorontsova N, et al., Expression of inducible inorganic carbon acquisition complexes is under the control of the FtsH protease in <i>Synechocystis</i> sp PCC 6803, Publisher: SPRINGER, Pages: 223-223, ISSN: 0166-8595
Hamilton M, Deak Z, Franco E, et al., Cytochrome <i>b</i>-559 is important for modulating electron transfer on the acceptor side of Photosystem II and for photoprotection during assembly of the Mn<sub>4</sub>Ca complex, Publisher: SPRINGER, Pages: 182-182, ISSN: 0166-8595
Nield J, Barker M, Boehm M, et al., Structural analysis of the FtsH2 homologue of <i>Synechocystis</i> sp PCC6803, Publisher: SPRINGER, Pages: 207-208, ISSN: 0166-8595
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