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1.
Sequencing and Modification of the Gene Encoding the 42-Kilodalton Protein in the Cytoplasmic Membrane of Synechococcus PCC 7942 总被引:4,自引:1,他引:3
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A 42-kilodalton cytoplasmic membrane protein is synthesized when high CO2-grown cells of Synechococcus PCC 7942 (Anacystis nidulans R2) are exposed to low CO2. The structural gene for this protein (cmpA) has been cloned and sequenced and shown to encode a 450 amino acid polypeptide with a molecular mass of 49 kilodalton. A deletion mutant lacking the 42-kilodalton protein was obtained by transformation of Synechococcus PCC 7942 following in vitro mutagenesis of the cloned gene. There were no significant differences between the mutant and wild-type cells in their growth rates under either low or high CO2 conditions. The activity of inorganic carbon (Ci) transport in the mutant was as high as that in the wild-type strain. In both types of cells, CO2 was the main species of Ci transported and the activities of CO2 and HCO3− transport increased when high CO2-grown cells were exposed to low CO2. We conclude that the 42-kilodalton protein is not directly involved in the Ci-accumulating mechanism of Synechococcus PCC 7942. 相似文献
2.
Synechococcus PCC 7942 contains two fructose-1,6-bisphosphataseisozymes (FBPase-I and FBPase-II), while Synechocystis PCC 6803has only one (FBPase-I) in spite of the occurrence of two FBPaseisozyme genes [Tamoi et al. (1998) Biochim. Biophys. Acta 1383:232]. We now demonstrate that disruption of the gene encodingFBPase-II (fbp-II) with a kanamycin resistance gene cartridgedoes not affect cell growth, Chl content, or CO2 assimilationin Synechococcus PCC 7942, and disruption of the gene encodingFBPase-I (fbp-I) is a lethal mutation in both cyanobacteria.Accordingly, it is clear that FBPase-I is necessary to sustainphotosynthesis and gluconeogenesis in cyanobacteria. (Received September 10, 1998; Accepted December 10, 1998) 相似文献
3.
4.
We have determined the complete nucleotide sequence of pAQ1,the smallest plasmid of the unicellular marine cyanobacteriumSynechococcus sp. PCC7002. The plasmid consists of 4,809 bpand has at least four open reading frames that potentially encodepolypeptides of 50 or more amino acids. We found that a palindromicelement, the core sequence of which is G(G/A)CGATCGCC, is over-representednot only in plasmid pAQ1 but also in the accumulated cyanobacterialgenomic sequences from Synechococcus sp. PCC6301, PCC7002, PCC7942,vulcanus and Synechocystis sp. PCC6803 within GenBank and EMBLdatabases. It suggests that this sequence might mediate generearrangement, thus increasing genetic diversity, since recombinationevents are frequent in cyanobacteria. 相似文献
5.
Sarah Joshua 《BBA》2005,1709(1):58-68
State transitions in cyanobacteria are a physiological adaptation mechanism that changes the interaction of the phycobilisomes with the Photosystem I and Photosystem II core complexes. A random mutagenesis study in the cyanobacterium Synechocystis sp. PCC6803 identified a gene named rpaC which appeared to be specifically required for state transitions. rpaC is a conserved cyanobacterial gene which was tentatively suggested to code for a novel signal transduction factor. The predicted gene product is a 9-kDa integral membrane protein. We have further examined the role of rpaC by overexpressing the gene in Synechocystis 6803 and by inactivating the ortholog in a second cyanobacterium, Synechococcus sp. PCC7942. Unlike the Synechocystis 6803 null mutant, the Synechococcus 7942 null mutant is unable to segregate, indicating that the gene is essential for cell viability in this cyanobacterium. The Synechocystis 6803 overexpressor is also unable to segregate, indicating that the cells can only tolerate a limited gene copy number. The non-segregated Synechococcus 7942 mutant can perform state transitions but shows a perturbed phycobilisome-Photosystem II interaction. Based on these results, we propose that the rpaC gene product controls the stability of the phycobilisome-Photosystem II supercomplex, and is probably a structural component of the complex. 相似文献
6.
The secY gene product is an essential component of the Escherichia coli cytoplasmic membrane, which mediates the protein translocation across the membrane. We found a gene homologous to secY in the genome of the cyanobacterium Synechococcus PCC7942. The deduced amino acid sequence, 439 amino acids long, shows 43% homology with that of the E. coli secY. The hydrophobic profile suggests that the Synechococcus SecY protein is an integral membrane protein containing ten membrane-spanning segments, which are closely related to the E. coli counterpart. The SecY protein may participate in the protein translocation across the cytoplasmic or thylakoid membrane in Synechococcus PCC7942. 相似文献
7.
Klaus-Peter Michel Pablo Exss-Sonne Gabriele Scholten-Beck Uwe Kahmann Hans Georg Ruppel Elfriede K. Pistorius 《Planta》1998,205(1):73-81
Iron-deficiency-induced protein A (IdiA) with a calculated molecular mass of 35 kDa has previously been shown to be essential
under manganese- and iron-limiting conditions in the cyanobacteria Synechococcus PCC 6301 and PCC 7942. Studies of mutants indicated that in the absence of IdiA mainly photosystem II becomes damaged, suggesting
that the major function of IdiA is in Mn and not Fe metabolism (Michel et al. 1996, Microbiology 142: 2635–2645). To further elucidate the function of IdiA, the immunocytochemical localization of IdiA
in the cell was examined. These investigations provided evidence that under mild Fe deficiency IdiA is intracellularly localized
and is mainly associated with the thylakoid membrane in Synechococcus PCC 6301. The protein became distributed throughout the cell under severe Fe limitation when substantial morphological changes
had already occurred. For additional verification of a preferential thylakoid membrane association of IdiA, these investigations
were extended to the thermophilic Synechococcus elongatus. In this cyanobacterium Mn deficiency could be obtained more rapidly than in the mesophilic Synechococcus PCC 6301 and PCC 7942, and the thylakoid membrane structure proved to be more stable under limiting growth conditions. The
immunocytochemical investigations with this cyanobacterium clearly supported a thylakoid membrane association of IdiA. In
addition, evidence was obtained for a localization of IdiA on the cytoplasmic side of the thylakoid membrane. All available
data support a function of IdiA as an Mn-binding protein that facilitates transport of Mn via the thylakoid membrane into
the lumen to provide photosystem II with Mn. A possible explanation for the observation that IdiA was not only expressed under
Mn deficiency but also under Fe deficiency is given in the discussion.
Received: 28 July 1997 / Accepted: 26 November 1997 相似文献
8.
Hideo Fukuda Miho Sakai Kazuhiro Nagahama Takao Fujii Masayoshi Matsuoka Yorinao Inoue Takahira Ogawa 《Biotechnology letters》1994,16(1):1-6
Summary Bioconversion of atmospheric carbon dioxide to ethylene was studied in a recombinant cyanobacterium. The gene for the ethylene-forming enzyme ofPseudomonas syringae pv.phaseolicola PK2 was cloned and expressed in the cyanobacteriumSynechococcus PCC7942 R2-SPc by use of a shuttle vector pUC303. The ethylene-forming activityin vivo ofSynechococcus PCC7942 R2-SPc that carried the gene for the ethylene-forming enzyme ofP. syringae pv.phaseolicola PK2 was one-fifth of that ofE. coli JM109 that harbored the same plasmid. The enzyme accounted for 0.021% by weight of the total soluble protein inSynechococcus PCC7942 R2-SPc. 相似文献
9.
The genome of Synechocystis sp. PCC 6803 contains an operon with homology to the sulfate permease of other prokaryotes. We used antibodies raised against cytoplasmic membrane protein to find three genes with strong homology to sbpA, orf81 and cysT genes of the cyanobacterium Synechococcus sp. PCC 7942, Escherichia coli, Salmonella typhymurium and Marchantia polymorpha. It is likely that the permease genes are expressed and the proteins are inserted into the cytoplasmic membrane. 相似文献
10.
Purification and Characterization of Phosphoribulokinase from the Cyanobacterium Synechococcus PCC7942 总被引:1,自引:0,他引:1
Wadano Akira; Kamata Yoichi; Iwaki Toshio; Nishikawa Keisuke; Hirahashi Tomohiro 《Plant & cell physiology》1995,36(7):1381-1385
Phosphoribulokinase (PRK) was purified to electrophoretic homogeneityfrom Synechococcus PCC7942 with high specific activity. Molecularmasses of the native enzyme and its subunit were 178 and 42kDa, respectively. Cys-17 and Cys-38 were conserved in the cyanobacterialPRK, but 18 amino acid residues between them were missing amongthe 40 residues found in higher plant PRKs. (Received February 1, 1995; Accepted July 27, 1995) 相似文献
11.
Synechococcus sp. strains PCC 7942 and PCC 6301 contain a 35 kDa protein called IdiA (Iron deficiency induced protein A) that is expressed in elevated amounts under Fe deficiency and to a smaller extent also under Mn deficiency. Absence of this
protein was shown to mainly damage Photosystem II. To decide whether IdiA has a function in optimizing and/or protecting preferentially
either the donor or acceptor side reaction of Photosystem II, a comparative analysis was performed of Synechococcus sp. PCC 7942 wild-type, the IdiA-free mutant, the previously constructed PsbO-free Synechococcus PCC 7942 mutant and a newly constructed Synechococcus PCC 7942 double mutant lacking both PsbO and IdiA. Measurements of the chlorophyll fluorescence and determinations of Photosystem
II activity using a variety of electron acceptors gave evidence that IdiA has its main function in protecting the acceptor
side of Photosystem II. Especially, the use of dichlorobenzoquinone, preferentially accepting electrons from QA, gave a decreased O2 evolving activity in the IdiA-free mutant. Investigations of the influence of hydrogen peroxide treatment on cells revealed
that this treatment caused a significantly higher damage of Photosystem II in the IdiA-free mutant than in wild-type. These
results suggest that although the IdiA protein is not absolutely required for Photosystem II activity in Synechococcus PCC 7942, it does play an important role in protecting the acceptor side against oxidative damage.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
12.
The gene products of sll0337 and slr0081 in Synechocystis sp. PCC 6803 have been identified as the homologues of the Escherichia coli phosphate-sensing histidine kinase PhoR and response regulator PhoB, respectively. Interruption of sll0337, the gene encoding the histidine protein kinase, by a spectinomycin-resistance cassette blocked the induction of alkaline phosphatase activity under phosphate-limiting conditions. A similar result was obtained when slr0081, the gene encoding the response regulator, was interrupted with a cassette conferring resistance to kanamycin. In addition, the phosphate-specific transport system was not up-regulated in our mutants when phosphate was limiting. Unlike other genes for bacterial phosphate-sensing two-component systems, sll0337 and slr0081 are not present in the same operon. Although there are three assignments for putative alkaline phosphatase genes in the Synechocystis sp. PCC 6803 genome, only sll0654 expression was detected by northern analysis under phosphate limitation. This gene codes for a 149 kDa protein that is homologous to the cyanobacterial alkaline phosphatase reported in Synechococcus sp. PCC 7942 [Ray, J.M., Bhaya, D., Block, M.A. and Grossman, A.R. (1991) J. Bact. 173: 4297–4309]. An alignment identified a conserved 177 amino acid domain that was found at the N-terminus of the protein encoded by sll0654 but at the C-terminus of the protein in Synechococcus sp. PCC 7942. 相似文献
13.
Sugita Mamoru; Luo Lijun; Ohta Masaru; Itadani Hiraku; Matsubayashi Tohru; Sugiura Masahiro 《DNA research》1995,2(2):71-76
A part of the tRNALeu (UAA) gene containing a 240-nucleotidegroup I intron was amplified by PCR from cyanobacterium SynechococcusPCC 6301 genomic DNA. The pre-tRNA synthesized from the clonedPCR product was efficiently self-spliced in vitro under physiologicalconditions. The gene encoding the tRNALeu (UAA), trnL-UAA, wasisolated from a Synechococcus PCC 6301 genomic library and thenucleotide sequence of a 2,167-bp portion was determined. ThetrnL-UAA consists of a 34-bp 5' exon, a 240-bp group I intronand a 50-bp 3' exon. In addition, three open reading frames(ORF1, ORF2 and ORF3) were found in the 5' and 3' flanking regionsof trnL-UAA. The predicted protein sequence of ORF3, which islocated 74-bp upstream from trnL-UAA on the opposite strand,shows 66.2% amino acid identity to that of the SynechocystisPCC 6803 gene encoding subunit L of NADH dehydrogenase (ndhL). 相似文献
14.
A cDNA encoding mouse metallothionein was cloned into the shuttle vector pUC303, creating a translational fusion with the bacterial chloramphenicol acetyltransferase gene. The resulting fusion protein has been expressed in the cyanobacteriumSynechococcus PCC7942. Cyanobacterial transformants expressed mouse metallothionein-specific mRNA species as detected by RNA slot blots. In addition, the transformants expressed a unique cadmium ionbinding protein corresponding to the predicted size of the mouse metallothionein fusion protein. Expression of this fusion protein conferred a two-to five-fold increase in cadmium ion tolerance and accumulation onSynechococcus PCC7942. 相似文献
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16.
A cyanobacterial expression vector was constructed using ribulose-1,5-bisphosphatecarboxylase/oxygenase (RuBisCO) promoter and terminator sequencesderived from Synechococcus PCC 6301. The recombinant plasmid,designated pARUB19, has an ampicillin-resistant (ApR) gene asa selectable marker and four unique restriction sites to allowthe insertion of foreign genes. Using this vector, the luciferasegene from the firefly, Photinus pyralis, was introduced intoSynechococcus PCC 6301 cells. The luciferase expression vectorcould be maintained stably in the host cells. Light productionof luciferin/luciferase was detected in the transformants. Luciferaseamounted to 1.2% of the total soluble protein. This plasmidmay facilitate higher levels of foreign gene expression in SynechococcusPCC 6301. 相似文献
17.
《Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression》1996,1305(1-2):19-24
The nucleotide sequence of the structural gene of nitrate reductase (narB) has been determined from the filamentous, non-heterocystous cyanobacterium Oscillatoria chalybea. The narB gene encodes a protein of 737 amino acid residues, which shows 61% identity to nitrate reductase of the unicellular cyanobacterium Synechococcus sp. PCC 7942 and only weak homologies to different bacterial molybdoenzymes, such as nitrate reductases or formate dehydrogenases. 相似文献
18.
A 1.2kb DNA fragment was cloned from Synechococcus sp. PCC7942, which is able phenotypicalty to complement a phoRcreC Escherichia coli mutant for the expression of alkaline phosphatase. A 2.5kb DNA fragment encompassing the putative gene was then cloned and its complete nucleotide sequence determined. Nucleotide sequencing revealed that the intact gene encodes a protein of 46389 Da, and that the deduced amino acid sequence shows a high degree of homology to those of the bacterial sensory kinase family. In the determined nucleotide sequence, another gene was adjacently located, which encodes a protein of 29012Da. This protein shows a high degree of homology to those of the response regulator family. Thus, we succeeded in the cloning of a pair of genes encoding the sensory kinase and response regulator, respectively, in a cyanobacterium. Mutant strains that lack these genes were constructed, and demonstrated to be defective in their ability to produce alkaline phosphatase and some inducible proteins in response to phosphate-limitation in the medium. These results imply that the gene products identified in this study are probably involved, either directly or indirectly, in the signal-transduction mechanism underlying regulation of the phosphate regulon in Synechococcus sp. PCC7942. Hence, the genes encoding the sensory kinase and response regulator were designated as sphS and sphR, respectively (S ynechococcusph osphate regulon). The SphS protein was demonstrated in vitro to undergo phosphorylation in the presence of ATP. 相似文献
19.
Maria Marjorette O. Peña William Burkhart George S. Bullerjahn 《Archives of microbiology》1995,163(5):337-344
A stable DNA/protein complex having an apparent molecular mass of approximately 150kDa was purified from nitrate-limited cultures of the cyanobacterium Synechococcus sp. strain PCC 7942. Amino-terminal peptide sequencing indicated that the polypeptide was structurally similar to the Dps protein of Escherichia coli; Dps is also known as the product of the starvation- and stationary-phase-inducible gene, pexB. The 150-kDa complex dissociated into a 22-kDa protein monomer after boiling in 2% SDS. The 150-kDa complex preparation had approximately a 10% nucleic acid content and upon dissociation released DNA fragments that were sensitive to S1 nuclease digestion. Immunoblot data indicated that the complex accumulates during stationary phase and during nitrogen, sulfur, and phosphorus limitation. DNA-binding assays indicated that the protein nonspecifically binds both linear and supercoiled DNA. Circular dichroism spectroscopy revealed that the Synechococcus sp. Dps-like protein contains extensive regions of alpha-helical secondary structure. We propose that the 150-kDa complex represents a hexameric aggregate of the Dps-like protein complexed with single-stranded DNA and serves to bind a portion of the chromosomal DNA under nutrient-limited conditions. 相似文献