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Schwabe, T. M.Gloddek, K.Schluesener, D.Kruip, J.
Purification of recombinant BtpA and Ycf3, proteins involved in membrane protein biogenesis in Synechocystis PCC 6803.
J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Mar 25;786(1-2):45-59.
PMID:12651001
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The gene products Ycf3 (hypothetical chloroplast open reading frame) and BtpA (biogenesis of thylakoid protein) are thought to be involved in the biogenesis of the membrane protein complex photosystem I (PSI) from Synechocystis PCC 6803. PSI consists of 12 different subunits and binds more than 100 cofactors, making it a model protein to study different aspects of membrane protein biogenesis. For a detailed biophysical characterization of Ycf3 and BtpA pure proteins must be available in sufficient quantities. Therefore we cloned the corresponding genes into expression vectors. To facilitate purification we created His-tagged versions of Ycf3 and BtpA in addition to the unmodified forms. Immobilized metal affinity chromatography (IMAC) yielded His-tagged proteins which were used for the production of antibodies. Purification strategies for non-tagged proteins could also be established: Ycf3 could be purified in soluble form using a two-step purification in which ammonium sulfate precipitation was combined with anion-exchange chromatography (IEC). BtpA had to be purified from inclusion bodies by two-consecutive IEC steps under denaturing conditions. An optimized refolding protocol was established that yielded pure BtpA. In all cases, MALDI-TOF peptide mass fingerprinting (PMF) was used to confirm protein identity. Initially, size exclusion chromatography and CD-spectroscopy were used for biophysical characterization of the proteins. Both Ycf3 and BtpA show homo-oligomerization in vitro. In summary, purification protocols for Ycf3 and BtpA have been designed that yield pure proteins which can be used to probe the molecular function of these proteins for membrane protein biogenesis.
MeSH terms
- *Bacterial Proteins
- Base Sequence
- Chromatography, Liquid/methods
- Circular Dichroism
- Cyanobacteria/*chemistry
- DNA Primers
- Electrophoresis, Polyacrylamide Gel
- Membrane Proteins/*biosynthesis/*isolation & purification
- Photosystem I Protein Complex/*isolation & purification
- Recombinant Proteins/isolation & purification
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
2
Qiao, J.Wang, J.Chen, L.Tian, X.Huang, S.Ren, X.Zhang, W.
Quantitative iTRAQ LC-MS/MS proteomics reveals metabolic responses to biofuel ethanol in cyanobacterial Synechocystis sp. PCC 6803.
J Proteome Res. 2012 Nov 2;11(11):5286-300. doi: 10.1021/pr300504w. Epub 2012 Oct 23.
PMID:23062023
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Recent progress in metabolic engineering has led to autotrophic production of ethanol in various cyanobacterial hosts. However, cyanobacteria are known to be sensitive to ethanol, which restricts further efforts to increase ethanol production levels in these renewable host systems. To understand the mechanisms of ethanol tolerance so that engineering more robust cyanobacterial hosts can be possible, in this study, the responses of model cyanobacterial Synechocystis sp. PCC 6803 to ethanol were determined using a quantitative proteomics approach with iTRAQ LC-MS/MS technologies. The resulting high-quality proteomic data set consisted of 24,887 unique peptides corresponding to 1509 identified proteins, a coverage of approximately 42% of the predicted proteins in the Synechocystis genome. Using a cutoff of 1.5-fold change and a p-value less than 0.05, 135 and 293 unique proteins with differential abundance levels were identified between control and ethanol-treated samples at 24 and 48 h, respectively. Functional analysis showed that the Synechocystis cells employed a combination of induced common stress response, modifications of cell membrane and envelope, and induction of multiple transporters and cell mobility-related proteins as protection mechanisms against ethanol toxicity. Interestingly, our proteomic analysis revealed that proteins related to multiple aspects of photosynthesis were up-regulated in the ethanol-treated Synechocystis cells, consistent with increased chlorophyll a concentration in the cells upon ethanol exposure. The study provided the first comprehensive view of the complicated molecular mechanisms against ethanol stress and also provided a list of potential gene targets for further engineering ethanol tolerance in Synechocystis PCC 6803.
MeSH terms
- *Biofuels
- Chlorophyll/metabolism
- Chromatography, Liquid/*methods
- Cyanobacteria/*metabolism
- Ethanol/*metabolism
- Flow Cytometry
- *Proteomics
- Synechocystis/*metabolism
- Tandem Mass Spectrometry/*methods
3
Wilde, A.Lunser, K.Ossenbuhl, F.Nickelsen, J.Borner, T.
Characterization of the cyanobacterial ycf37: mutation decreases the photosystem I content.
Biochem J. 2001 Jul 1;357(Pt 1):211-6.
PMID:11415451
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We have constructed and analysed a cyanobacterial mutant that lacks the putative homologue of ycf37, the chloroplast open reading frame 37, which is conserved in different algae, but missing in the plastome of higher plants. In this report we show that Ycf37 of Synechocystis sp. PCC 6803 contains three tetratrico-peptide repeat (TPR) units resembling the structural organization of Ycf3, a protein that has been suggested to function as a chaperone during photosystem (PS) I complex formation. We demonstrate a light-activated transcript accumulation of this gene. Inactivation of ycf37 leads to a lower PSI/PSII ratio and a higher phycocyanin/chlorophyll ratio in Synechocystis cells. The observed alterations in the ycf37 mutants and the structural organization of the gene product suggest a functional role in PSI stability or assembly.
MeSH terms
- Amino Acid Sequence
- *Bacterial Proteins
- Consensus Sequence
- Cyanobacteria/*genetics/*metabolism
- DNA, Bacterial/chemistry/genetics
- Genes, Bacterial
- Light-Harvesting Protein Complexes
- Molecular Sequence Data
- *Mutagenesis
- *Open Reading Frames
- Oxygen/metabolism
- Photosynthetic Reaction Center Complex Proteins/chemistry/*genetics/*metabolism
- *Photosystem I Protein Complex
- Repetitive Sequences, Nucleic Acid
- Sequence Alignment
- Sequence Homology, Amino Acid
- Spectrometry, Fluorescence
4
Schmitt, W. A. JrRaab, R. M.Stephanopoulos, G.
Elucidation of gene interaction networks through time-lagged correlation analysis of transcriptional data.
Genome Res. 2004 Aug;14(8):1654-63.
PMID:15289483
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The photosynthetic cyanobacterium Synechocystis sp. strain PCC 6803 uses a complex genetic program to control its physiological response to alternating light conditions. To study this regulatory program time-series experiments were conducted by exposing Synechocystis sp. to serial perturbations in light intensity. In each experiment whole-genome DNA microarrays were used to monitor gene transcription in 20-min intervals over 8- and 16-h periods. The data was analyzed using time-lagged correlation analysis, which identifies genetic interaction networks by constructing correlations between time-shifted transcription profiles with different levels of statistical confidence. These networks allow inference of putative cause-effect relationships among the organism's genes. Using light intensity as our initial input signal, we identified six groups of genes whose time-lagged profiles possessed significant correlation, or anti-correlation, with the light intensity. We expanded this network by using the average profile from each group of genes as a seed, and searching for other genes whose time-lagged profiles possessed significant correlation, or anti-correlation, with the group's average profile. The final network comprised 50 different groups containing 259 genes. Several of these gene groups possess known light-stimulated gene clusters, such as Synechocystis sp. photosystems I and II and carbon dioxide fixation pathways, while others represent novel findings in this work.
MeSH terms
- Cyanobacteria/*genetics
- *Gene Expression Profiling
- Gene Expression Regulation, Bacterial
- *Genes, Bacterial
- *Genome, Bacterial
- Light
- Models, Biological
- Oligonucleotide Array Sequence Analysis/methods
- Time Factors
5
Duhring, U.Irrgang, K. D.Lunser, K.Kehr, J.Wilde, A.
Analysis of photosynthetic complexes from a cyanobacterial ycf37 mutant.
Biochim Biophys Acta. 2006 Jan;1757(1):3-11. Epub 2005 Dec 1.
PMID:16364235
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The Ycf37 protein has been suggested to be involved in the biogenesis and/or stability of the cyanobacterial photosystem I (PSI). With Ycf37 specific antibodies, we analyzed the localization of Ycf37 within the thylakoid membranes of the cyanobacterium Synechocystis sp. PCC 6803. Inspection of a sucrose gradient profile indicated that small amounts of Ycf37 co-fractionated with monomeric photosynthetic complexes, but not with trimeric PSI. Isolating 3xFLAG epitope-tagged Ycf37 by affinity-tag purification rendered several PSI subunits that specifically co-precipitated with this protein. Blue-native PAGE newly revealed two monomeric PSI complexes (PSI and PSI*) in wild-type thylakoids. The lower amount of PsaK present in PSI* may explain its higher electrophoretic mobility. PSI* was more prominent in high-light grown cells and interestingly proved absent in the Deltaycf37 mutant. PSI* appeared again when the mutant was complemented in trans with the wild-type ycf37 gene. In the Deltaycf37 mutant the amount of trimeric PSI complexes was reduced to about 70% of the wild-type level with no significant changes in photochemical activity and subunit composition of the remaining photosystems. Our results indicate that Ycf37 plays a specific role in the preservation of PSI* and the biogenesis of PSI trimers.
MeSH terms
- Bacterial Proteins/*analysis/genetics/*metabolism
- Electrophoresis, Polyacrylamide Gel
- Gene Deletion
- Immunoprecipitation
- Photosystem I Protein Complex/*analysis/genetics/*metabolism
- Protein Subunits/analysis/genetics/metabolism
- Synechocystis/*enzymology/genetics
- Thylakoids/*enzymology
6
Zak, E.Norling, B.Maitra, R.Huang, F.Andersson, B.Pakrasi, H. B.
The initial steps of biogenesis of cyanobacterial photosystems occur in plasma membranes.
Proc Natl Acad Sci U S A. 2001 Nov 6;98(23):13443-8. Epub 2001 Oct 30.
PMID:11687660
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During oxygenic photosynthesis in cyanobacteria and chloroplasts of plants and eukaryotic algae, conversion of light energy to biologically useful chemical energy occurs in the specialized thylakoid membranes. Light-induced charge separation at the reaction centers of photosystems I and II, two multisubunit pigment-protein complexes in the thylakoid membranes, energetically drive sequential photosynthetic electron transfer reactions in this membrane system. In general, in the prokaryotic cyanobacterial cells, the thylakoid membrane is distinctly different from the plasma membrane. We have recently developed a two-dimensional separation procedure to purify thylakoid and plasma membranes from the genetically widely studied cyanobacterium Synechocystis sp. PCC 6803. Immunoblotting analysis demonstrated that the purified plasma membrane contained a number of protein components closely associated with the reaction centers of both photosystems. Moreover, these proteins were assembled in the plasma membrane as chlorophyll-containing multiprotein complexes, as evidenced from nondenaturing green gel and low-temperature fluorescence spectroscopy data. Furthermore, electron paramagnetic resonance spectroscopic analysis showed that in the partially assembled photosystem I core complex in the plasma membrane, the P700 reaction center was capable of undergoing light-induced charge separation. Based on these data, we propose that the plasma membrane, and not the thylakoid membrane, is the site for a number of the early steps of biogenesis of the photosynthetic reaction center complexes in these cyanobacterial cells.
MeSH terms
- Cell Membrane/metabolism
- Cyanobacteria/*metabolism
- Electron Spin Resonance Spectroscopy
- Membrane Proteins/*biosynthesis
- Photosynthetic Reaction Center Complex Proteins/*biosynthesis
- Spectrometry, Fluorescence
7
Churin, Y. N.Shalak, I. N.Borner, T.Shestakov, S. V.
Physical and genetic map of the chromosome of the unicellular cyanobacterium Synechocystis sp. strain PCC 6803.
J Bacteriol. 1995 Jun;177(11):3337-43.
PMID:7768838
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A combined physical and genetic map of the cyanobacterium Synechocystis sp. strain PCC 6803 chromosome was constructed. An estimated genome size of 3.82 Mb was obtained by summing the sizes of 25 MluI or 40 NotI fragments seen by pulsed-field electrophoresis. The order of the restriction fragments was determined by using two independent experimental approaches: pulsed-field fragment hybridization and linking clone analysis. The relative positions of 30 known genes or gene clusters were localized.
MeSH terms
- Cloning, Molecular
- Cyanobacteria/*genetics
- *Genes
- *Restriction Mapping
8
Yokouchi, H.Fukuoka, Y.Mukoyama, D.Calugay, R.Takeyama, H.Matsunaga, T.
Whole-metagenome amplification of a microbial community associated with scleractinian coral by multiple displacement amplification using phi29 polymerase.
Environ Microbiol. 2006 Jul;8(7):1155-63.
PMID:16817924
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Limitations in obtaining sufficient specimens and difficulties in extracting high quality DNA from environmental samples have impeded understanding of the structure of microbial communities. In this study, multiple displacement amplification (MDA) using phi29 polymerase was applied to overcome these hindrances. Optimization of the reaction conditions for amplification of the bacterial genome and evaluation of the MDA product were performed using cyanobacterium Synechocystis sp. strain PCC6803. An 8-h MDA reaction yielded a sufficient quantity of DNA from an initial amount of 0.4 ng, which is equivalent to approximately 10(5) cells. Uniform amplification of genes randomly selected from the cyanobacterial genome was confirmed by real-time polymerase chain reaction. The metagenome from bacteria associated with scleractinian corals was used for whole-genome amplification using phi29 polymerase to analyse the microbial diversity. Unidentified bacteria with less than 93% identity to the closest 16S rDNA sequences deposited in DNA Data Bank of Japan were predominantly detected from the coral-associated bacterial community before and after the MDA procedures. Sequencing analysis indicated that alpha-Proteobacteria was the dominant group in Pocillopora damicornis. This study demonstrates that MDA techniques are efficient for genome wide investigation to understand the actual microbial diversity in limited bacterial samples.
MeSH terms
- Animals
- Anthozoa/*microbiology
- Bacillus Phages/enzymology
- Bacteriological Techniques/*methods
- Biodiversity
- DNA Primers
- DNA-Directed DNA Polymerase/*genetics
- Ecosystem
- Gene Library
- Molecular Sequence Data
- Nucleic Acid Amplification Techniques/*methods
- Phylogeny
- RNA, Ribosomal, 16S/*genetics
9
Duhring, U.Ossenbuhl, F.Wilde, A.
Late assembly steps and dynamics of the cyanobacterial photosystem I.
J Biol Chem. 2007 Apr 13;282(15):10915-21. Epub 2007 Feb 15.
PMID:17303568
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The dynamics of photosystem I assembly in cyanobacteria have been addressed using in vivo pulse-chase labeling of Synechocystis sp. PCC 6803 proteins in combination with blue native polyacrylamide gel electrophoresis. The analyses indicate the existence of three different monomeric photosystem I complexes and also the high stability of photosystem I trimers. We show that in addition to a complete photosystem I monomer, containing all 11 subunits, we detected a PsaK-less monomer and a short-lived PsaL/PsaK-less complex. The latter two monomers were missing in the ycf37 mutant of Synechocystis sp. PCC 6803 that accumulates also less trimers. Pulse-chase experiments suggest that the three monomeric complexes have different functions in the biogenesis of the trimer. Based on these findings we propose a model where PsaK is incorporated in the latest step of photosystem I assembly. The PsaK-less photosystem I monomer may represent an intermediate complex that is important for the exchange of the two PsaK variants during high light acclimation. Implications of the presented data with respect to Ycf37 function are discussed.
MeSH terms
- Dimerization
- Light
- Mutation/genetics
- Photosystem I Protein Complex/genetics/*metabolism
- Protein Binding
- Protein Subunits/genetics/metabolism
- Synechocystis/*enzymology/genetics
- Thylakoids/metabolism
10
Singh, A. K.Elvitigala, T.Bhattacharyya-Pakrasi, M.Aurora, R.Ghosh, B.Pakrasi, H. B.
Integration of carbon and nitrogen metabolism with energy production is crucial to light acclimation in the cyanobacterium Synechocystis.
Plant Physiol. 2008 Sep;148(1):467-78. doi: 10.1104/pp.108.123489. Epub 2008 Jul 3.
PMID:18599646
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Light drives the production of chemical energy and reducing equivalents in photosynthetic organisms required for the assimilation of essential nutrients. This process also generates strong oxidants and reductants that can be damaging to the cellular processes, especially during absorption of excess excitation energy. Cyanobacteria, like other oxygenic photosynthetic organisms, respond to increases in the excitation energy, such as during exposure of cells to high light (HL) by the reduction of antenna size and photosystem content. However, the mechanism of how Synechocystis sp. PCC 6803, a cyanobacterium, maintains redox homeostasis and coordinates various metabolic processes under HL stress remains poorly understood. In this study, we have utilized time series transcriptome data to elucidate the global responses of Synechocystis to HL. Identification of differentially regulated genes involved in the regulation, protection, and maintenance of redox homeostasis has offered important insights into the optimized response of Synechocystis to HL. Our results indicate a comprehensive integrated homeostatic interaction between energy production (photosynthesis) and energy consumption (assimilation of carbon and nitrogen). In addition, measurements of physiological parameters under different growth conditions showed that integration between the two processes is not a consequence of limitations in the external carbon and nitrogen levels available to the cells. We have also discovered the existence of a novel glycosylation pathway, to date known as an important nutrient sensor only in eukaryotes. Up-regulation of a gene encoding the rate-limiting enzyme in the hexosamine pathway suggests a regulatory role for protein glycosylation in Synechocystis under HL.
MeSH terms
- *Acclimatization
- Carbon Dioxide/*metabolism
- Cluster Analysis
- Energy Metabolism
- Gene Expression Profiling
- Genes, Bacterial
- Hexosamines/metabolism
- Homeostasis
- *Light
- Nitrogen/*metabolism
- Oligonucleotide Array Sequence Analysis
- Oxidation-Reduction
- Photosynthesis
- Signal Transduction
- Synechocystis/genetics/*metabolism
11
Schriek, S.Aguirre-von-Wobeser, E.Nodop, A.Becker, A.Ibelings, B. W.Bok, J.Staiger, D.Matthijs, H. C.Pistorius, E. K.Michel, K. P.
Transcript profiling indicates that the absence of PsbO affects the coordination of C and N metabolism in Synechocystis sp. PCC 6803.
Physiol Plant. 2008 Jul;133(3):525-43. doi: 10.1111/j.1399-3054.2008.01119.x. Epub 2008 Jul 1.
PMID:18419737
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Transcript profiling of nitrate-grown Synechocystis sp. PCC 6803 PsbO-free mutant cells in comparison to wild-type (WT) detected substantial deviations. Because we had previously observed phenotypical differences between Synechocystis sp. PCC 6803 WT and its corresponding PsbO-free mutant when cultivated with l-arginine as sole N source and a light intensity of 200 mumol photons m(-2) s(-1), we also performed transcript profiling for both strains grown either with nitrate or with l-arginine as sole N source. We observed a total number of 520 differentially regulated transcripts in Synechocystis WT because of a shift from nitrate- to l-arginine-containing BG11 medium, while we detected only 13 differentially regulated transcripts for the PsbO-free mutant. Thus, the PsbO-free Synechocystis mutant had already undergone a preconditioning process for growth with l-arginine in comparison to WT. While Synechocystis WT suffered from growth with l-arginine at a light intensity of 200 mumol photons m(-2) s(-1), the PsbO-free mutant developed only a minor stress phenotype. In summary, our results suggest that the absence of PsbO in Synechocystis affects the coordination of photosynthesis/respiration and l-arginine metabolism through complex probably redox-mediated regulatory pathways. In addition, we show that a comparison of the transcriptomes of nitrate-grown Synechococcus elongatus PCC 7942 WT cells and its corresponding PsbO-free mutant cells resulted in only a few differentially regulated transcripts between both strains. The absence of the manganese/calcium-stabilizing PsbO protein of PSII with an assigned regulatory function for photosynthetic water oxidation causes bigger changes in the transcriptome of the permissive photoheterotrophically growing Synechocystis sp. PCC 6803 than in the transcriptome of the obligate photoautotrophically growing S. elongatus PCC 7942.
MeSH terms
- Arginine/pharmacology
- Carbon/*metabolism
- Gene Expression Profiling/*methods
- Gene Expression Regulation, Bacterial/drug effects
- Nitrates/pharmacology
- Nitrogen/*metabolism
- Oligonucleotide Array Sequence Analysis
- Photosystem II Protein Complex/genetics/*metabolism
- Synechocystis/drug effects/*genetics/metabolism
12
Wang, Y.Xu, W.Chitnis, P. R.
Identification and bioinformatic analysis of the membrane proteins of synechocystis sp. PCC 6803.
Proteome Sci. 2009 Mar 25;7:11. doi: 10.1186/1477-5956-7-11.
PMID:19320970
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BACKGROUND: The membranes of Synechocystis sp. PCC 6803 play a central role in photosynthesis, respiration and other important metabolic pathways. Comprehensive identification of the membrane proteins is of importance for a better understanding of the diverse functions of its unique membrane structures. Up to date, approximately 900 known or predicted membrane proteins, consisting 24.5% of Synechocystis sp. PCC 6803 proteome, have been indentified by large-scale proteomic studies. RESULTS: To resolve more membrane proteins on 2-D gels for mass spectrometry identification, we separated integral proteins from membrane associated proteins and collected them as the integral and peripheral fractions, respectively. In total, 95 proteins in the peripheral fraction and 29 proteins in the integral fraction were identified, including the 5 unique proteins that were not identified by any previous studies. Bioinformatic analysis revealed that the identified proteins can be functionally classified into 14 distinct groups according to the cellular functions annotated by Cyanobase, including the two largest groups hypothetical and unknown, and photosynthesis and respiration. Homology analysis indicates that the identified membrane proteins are more conserved than the rest of the proteome. CONCLUSION: The proteins identified in this study combined with other published proteomic data provide the most comprehensive Synechocystis proteome catalog, which will serve as a useful reference for further detailed studies to address protein functions through both traditional gene-by-gene and systems biology approaches.
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Huang, F.Fulda, S.Hagemann, M.Norling, B.
Proteomic screening of salt-stress-induced changes in plasma membranes of Synechocystis sp. strain PCC 6803.
Proteomics. 2006 Feb;6(3):910-20.
PMID:16400685
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The plasma membrane of a cyanobacterial cell is crucial as barrier against the outer medium. It is also an energy-transducing membrane as well as essential for biogenesis of cyanobacterial photosystems and the endo-membrane system. Previously we have identified 57 different proteins in the plasma membrane of control cells from Synechocystis sp. strain PCC6803. In the present work, proteomic screening of salt-stress proteins in the plasma membrane resulted in identification of 109 proteins corresponding to 66 different gene products. Differential and quantitative analyses of 2-DE profiles of plasma membranes isolated from both control and salt-acclimated cells revealed that twenty proteins were enhanced/induced and five reduced during salt stress. More than half of the enhanced/induced proteins were periplasmic binding proteins of ABC-transporters or hypothetical proteins. Proteins that exhibited the highest enhancement during salt stress include FutA1 (Slr1295) and Vipp1 (Sll0617), which have been suggested to be involved in protection of photosystem II under iron deficiency and in thylakoid membrane formation, respectively. Other salt-stress proteins were regulatory proteins such as PII protein, LrtA, and a protein that belongs to CheY subfamily. The physiological significance of the identified salt-stress proteins in the plasma membrane is discussed integrating our current knowledge on cyanobacterial stress physiology.
MeSH terms
- Bacterial Proteins/*metabolism
- Cell Membrane/*metabolism
- Electrophoresis, Gel, Two-Dimensional
- Membrane Proteins/*metabolism
- *Proteome
- Sodium Chloride/*pharmacology
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
- Subcellular Fractions/metabolism
- Synechocystis/drug effects/growth & development/*metabolism
14
Peter, E.Wallner, T.Wilde, A.Grimm, B.
Comparative functional analysis of two hypothetical chloroplast open reading frames (ycf) involved in chlorophyll biosynthesis from Synechocystis sp. PCC6803 and plants.
J Plant Physiol. 2011 Aug 15;168(12):1380-6. doi: 10.1016/j.jplph.2011.01.014. Epub 2011 Mar 8.
PMID:21388705
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Hypothetical chloroplast open reading frames (ycfs) are highly conserved and interspecifically occurring genes in plastomes of plants and algae with significant functions in gene expression and photosynthesis. However, the function of many ycfs is still in vain so that attention is directed to other chloroplast functions such as metabolism of co-factors, protein translocation and protection against abiotic stress. We provide a comprehensive functional description of ycf53 and ycf59, two genes involved in chlorophyll biosynthesis. While ycf59 encodes an essential enzymatic component of Mg protoporphyrin monomethylester cyclase, ycf53 encodes a posttranslational regulator of chlorophyll biosynthesis. Their roles in tetrapyrrole biosynthesis were compared by using cyanobacterial and plant mutants with modulated expression of these two genes. Our work provides indications for diverse effects of these homologous gene products in plants and cyanobacteria on tetrapyrrole biosynthesis and photosynthesis.
MeSH terms
- Biosynthetic Pathways
- Chlorophyll/*biosynthesis
- Chloroplasts/*genetics
- Esters/metabolism
- Gene Knockout Techniques
- Models, Biological
- Open Reading Frames/*genetics
- Phylogeny
- Plant Proteins/chemistry/metabolism
- Plants/enzymology/*genetics
- Protein Subunits/metabolism
- Protoporphyrins/metabolism
- Synechocystis/*genetics
- Tetrapyrroles/biosynthesis
15
Zhang, S.Laborde, S. M.Frankel, L. K.Bricker, T. M.
Four novel genes required for optimal photoautotrophic growth of the cyanobacterium Synechocystis sp. strain PCC 6803 identified by in vitro transposon mutagenesis.
J Bacteriol. 2004 Feb;186(3):875-9.
PMID:14729717
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Four novel Synechocystis sp. strain PCC 6803 genes (sll1495, sll0804, slr1306, and slr1125) which encode hypothetical proteins were determined by transposon mutagenesis to be required for optimal photoautotrophic growth. Mutations were also recovered in ccmK4, a carboxysome coat protein homologue, and me, the decarboxylating NADP(+)-dependent malic enzyme. This is the first report that these known genes are required for optimal photoautotrophy.
MeSH terms
- Cyanobacteria/*genetics/*growth & development
- *DNA Transposable Elements
- Mutagenesis, Insertional
- *Photosynthesis
16
Sato, S.Shimoda, Y.Muraki, A.Kohara, M.Nakamura, Y.Tabata, S.
A large-scale protein protein interaction analysis in Synechocystis sp. PCC6803.
DNA Res. 2007 Oct 31;14(5):207-16. Epub 2007 Nov 13.
PMID:18000013
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Protein-protein interactions (PPIs) play crucial roles in protein function for a variety of biological processes. Data from large-scale PPI screening has contributed to understanding the function of a large number of predicted genes from fully sequenced genomes. Here, we report the systematic identification of protein interactions for the unicellular cyanobacterium Synechocystis sp. strain PCC6803. Using a modified high-throughput yeast two-hybrid assay, we screened 1825 genes selected primarily from (i) genes of two-component signal transducers of Synechocystis, (ii) Synechocystis genes whose homologues are conserved in the genome of Arabidopsis thaliana, and (iii) genes of unknown function on the Synechocystis chromosome. A total of 3236 independent two-hybrid interactions involving 1920 proteins (52% of the total protein coding genes) were identified and each interaction was evaluated using an interaction generality (IG) measure, as well as the general features of interacting partners. The interaction data obtained in this study should provide new insights and novel strategies for functional analyses of genes in Synechocystis, and, additionally, genes in other cyanobacteria and plant genes of cyanobacterial origin.
MeSH terms
- Arabidopsis/genetics/metabolism
- Arabidopsis Proteins/genetics/metabolism
- Bacterial Proteins/*genetics/*metabolism
- Base Sequence
- Conserved Sequence
- DNA Primers/genetics
- DNA, Bacterial/genetics
- Genome, Bacterial
- Genome, Plant
- *Protein Interaction Mapping/statistics & numerical data
- Signal Transduction/genetics
- Species Specificity
- Synechocystis/*genetics/*metabolism
- Two-Hybrid System Techniques
0.747082203 [0.208528734, 0.396730974, 0.599513075, 0.679471326, 0.742138128]
