Inorganic carbon levels regulate growth via SigC signaling cascade in cyanobacteria
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PROVIDER: PRJNA976260 | ENA |
REPOSITORIES: ENA
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Project description:Gene expression changes were followed in cultures of the cyanobacterium Synechocystis sp. PCC 6803 substrain GT-T cultivated at ambient air or supplemented with 3% CO2. The acclimation to different CO2 concentrations is crucial for photoautotrophic organisms living in aquatic environments such as cyanobacteria. Samples were taken before and 1 h and 24 h after transfer to the3 % CO2 environment. The analyzed strains were wild type, a deletion mutant of gene ssl2982/rpoZ (ΔrpoZ) and two suppressor strains (R1, ΔrpoZ-S1 and R2, ΔrpoZ-S2). In cyanobacteria, elevated CO2 is known to down-regulate carbon concentrating mechanisms and accelerate photosynthesis and growth, but mechanism(s) of carbon signalling remains only poorly understood. Here we reveal a novel signalling cascade connecting the amount of CO2 and growth in the model cyanobacterium Synechocystis sp. PCC 6803. Deletion of the small ω subunit of the RNA polymerase (RNAP) in the ΔrpoZ strain prevents normal high-CO2-induced up-regulation of numerous photosynthetic genes, and low expression of peptidoglycan synthesis genes induced lysis of dividing ΔrpoZ cells in high CO2. Spontaneously raised secondary mutations in the ssr1600 gene rescued the high-CO2-sensitive phenotype of the ΔrpoZ strain. Biochemical analyses showed that the ssr1600 gene encodes an anti-σ factor antagonist of group 2 σ factor SigC, and 3D structural modelling suggest that Slr1861 functions as an anti-SigC factor. In ΔrpoZ, excess formation of RNAP-SigC lead to high CO2 sensitive phenotype, whereas the drastically reduced Ssr1600 content in the suppressor mutants reduce the formation of the RNAP-SigC holoenzyme to the similar level as in the control strain, allowing almost normal transcriptome and growth of suppressor lines in high CO2. We propose that the SigC σ factor, the anti-SigC factor Slr1861 and the anti-SigC antagonist Ssr1600 forms a growth regulating signalling cascade in cyanobacteria.
2025-05-01 | GSE233434 | GEO
Project description:CO2-concentrating mechanism (CCM) is a unique adaptation that evolved in cyanobacteria for inorganic carbon (Ci) sequestration under limiting Ci levels. The CCM is based on several Ci uptake systems and the carboxysome in which RubisCO is confined in a high CO2 environment. Many CCM components are transcriptionally upregulated under low Ci levels through the combined action of the transcriptional factors NdhR, CmpR and the cAMP-binding SyCRP1. Here we show that, SyCRP1 senses the high Ci levels through high cAMP thereby repressing CCM genes. SyCRP1 possess low- and high-affinity cAMP binding sites. Further, the specific interaction of SyCRP1 with membrane lipids was shown. Flag-tagged SyCRP1, when ectopically expressed, was localized to the membrane and released to the cytosol upon cAMP binding under high Ci levels. Finally, ChIP-seq experiments revealed that SyCRP1 binds cis-acting elements at the promoters of multiple transcription units, mainly those involved in CCM. EMSA confirmed SyCRP1’s binding to target DNA-elements. Collectively, our results establish SyCRP1 as a transcriptional repressor of the cyanobacterial CCM under high Ci conditions.
2025-10-05 | GSE295773 | GEO
Project description:The cAMP receptor protein, SyCRP1 acts as a transcriptional repressor of CO2-concentrating mechanism genes at high inorganic carbon levels in cyanobacteria
| PRJNA1256339 | ENA
Project description:In eubacteria, replacement of one σ factor in the RNA polymerase (RNAP) holoenzyme by another one changes the transcription pattern. Cyanobacteria are eubacteria characterized by oxygenic photosynthesis and they typically encode numerous group 2 σ factors that closely resemble the essential primary σ factor. A mutant strain of the model cyanobacterium Synechocystis sp. PCC 6803 without functional group 2 σ factors (named as ΔsigBCDE) could not acclimate to heat, high salt, or bright light stress but in standard conditions ΔsigBCDE grew only 9% slower than the control strain. One-fifth of the genes in ΔsigBCDE were differently expressed compared to the control strain in standard growth conditions and several physiological changes in photosynthesis, and pigment and lipid compositions were detected. To directly analyze the σ factor content of RNAP holoenzyme in vivo, a His-tag was added to the γ subunit of RNAP in Synechocystis and RNAPs were collected. The results revealed that all group 2 σ factors were recruited by RNAP in standard conditions, but recruitment of SigB and SigC increased in heat stress, SigD in bright light, SigE in darkness and SigB, SigC and SigE in high salt, explaining the poor acclimation of ΔsigBCDE to these stress conditions. Cells from cyanobacteria Synechocystis sp. PCC 6803 named as control strain (CS) and a mutant strain without any functional group 2 sigma factors, ΔsigBCDE, were harvested (A730=1, 40 mL) directly from standard growth conditions (continuous illumination at the PPFD of 40 µmol m-2s-1, 32°C, ambient CO2). From three to four independent experiments were performed at each conditions.
2015-09-15 | E-GEOD-69981 | biostudies-arrayexpress
Project description:Microbial samples affected by inorganic carbon Genome sequencing and assembly
| PRJNA863027 | ENA
Project description:Thioredoxins play an essential role in regulating enzyme activity upon response to environmental changes, especially in photosynthetic organisms. These are crucial for the regulation of metabolism in cyanobacteria, but the key central processes redox-regulated remain to be determined. Physiological, metabolic and transcriptomic characterization of a conditional mutant of the essential trxA gene in Synechocystis (STXA2) revealed that reduced TrxA levels alter cell morphology and enter in a dormant-like state. TrxA depletion in the STXA2 strain inhibited protein synthesis and led to changes in amino acid pools and nitrogen/carbon reserve polymers, accompanied by oxidation of the Elongation Factor EF-Tu. Transcriptomic analysis of TrxA depletion in STXA2 showed a robust transcriptional response. Genes that responded negatively were grouped in a large cluster and were directly related to photosynthesis, ATP synthesis and CO2 fixation, while genes that responded positively were grouped in different clusters and related to respiratory electron transport, carotenoids synthesis, amino acid metabolism and protein degradation, among others. Regulation of cyanobacterial metabolism is complex and fine-tuned, our results link TrxA to the switch from anabolic to maintenance metabolism in cyanobacteria, regulating carbon and nitrogen balance.
2023-09-24 | GSE243518 | GEO
Project description:Nostoc cyanobacteria are capable to form symbiotic relationships with plants, transitioning to a heterotrophic lifestyle in return for providing bioavailable nitrogen to the host. The diazotrophic photoautotrophs also serve as a hub for a specialized heterotrophic bacterial community whose physiological contributions are poorly understood. By comparing the axenic strain N. punctiforme PCC 73102 and the related strains Nostoc sp. KVJ2 and KVJ3, which still maintain their heterotrophic microbiome, we were able to demonstrate an almost obligate dependence of the cyanobacteria on the heterotrophic partners under carbon-limiting conditions. Detailed analysis of the intimate bilateral relationship between Nostoc punctiforme and the isolate Agrobacterium tumefaciens Het4 using multi-omics technologies and microscopy uncovered a complex partnership characterized, among other traits, by competition for iron and facilitation for carbon. Although competitive interactions with A. tumefaciens Het4 compromise nitrogen fixation and stimulate the degradation of cyanophycin, mutualistic dependency prevails under inorganic carbon limitation. Both the absence of the high affinity bicarbonate uptake transporter SbtA and the prevalent extracarboxysomal localization of the carbon-fixing enzyme RubisCO as detected by immunofluorescence microscopy suggest a weak carbon concentrating mechanism in N. punctiforme that enforces a dependence on heterotrophic bacteria. Further, immunofluorescence, electron microscopic and proteomic analyses reveal a pronounced extracellular recycling of proteins under N- and C-limiting conditions. The pivotal influence of heterotrophic bacteria on symbiotic Nostoc strains should be considered when analyzing these strains, especially in the free-living state, and also sheds new light on the benefit to Nostoc of the provision of organic carbon by plant hosts.
2025-04-09 | PXD052511 | Pride
Project description:Regulation of CO2 fixation in cyanobacteria is important both for the organism and the global carbon balance. Here we show that phosphoketolase in Synechococcus elongatus PCC7942 (SeXPK) possesses a distinct ATP sensing mechanism, which upon ATP drops, allows SeXPK to divert precursors of the RuBisCO substrate away from the Calvin-Benson-Bassham (CBB) cycle. Deleting the SeXPK gene increased CO2 fixation particularly during light-dark transitions. In high-density cultures, the xpk strain showed a 60% increase in carbon fixation, and unexpectedly resulted in sucrose secretion without any pathway engineering. Using cryo-EM analysis, we discovered that these functions were enabled by a unique allosteric regulatory site involving two subunits jointly binding two ATP, which constantly suppresses the activity of SeXPK until the ATP level drops. This magnesium-independent ATP allosteric site is present in many species across all three domains of life, where it may also play important regulatory functions.
2023-08-08 | GSE227397 | GEO
Project description:In eubacteria, replacement of one σ factor in the RNA polymerase (RNAP) holoenzyme by another one changes the transcription pattern. Cyanobacteria are eubacteria characterized by oxygenic photosynthesis and they typically encode numerous group 2 σ factors that closely resemble the essential primary σ factor. A mutant strain of the model cyanobacterium Synechocystis sp. PCC 6803 without functional group 2 σ factors (named as ΔsigBCDE) could not acclimate to heat, high salt, or bright light stress but in standard conditions ΔsigBCDE grew only 9% slower than the control strain. One-fifth of the genes in ΔsigBCDE were differently expressed compared to the control strain in standard growth conditions and several physiological changes in photosynthesis, and pigment and lipid compositions were detected. To directly analyze the σ factor content of RNAP holoenzyme in vivo, a His-tag was added to the γ subunit of RNAP in Synechocystis and RNAPs were collected. The results revealed that all group 2 σ factors were recruited by RNAP in standard conditions, but recruitment of SigB and SigC increased in heat stress, SigD in bright light, SigE in darkness and SigB, SigC and SigE in high salt, explaining the poor acclimation of ΔsigBCDE to these stress conditions.
2015-09-15 | GSE69981 | GEO