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1.
Cell ; 178(2): 374-384.e15, 2019 07 11.
Artigo em Inglês | MEDLINE | ID: mdl-31299201

RESUMO

Multicellular lifestyle requires cell-cell connections. In multicellular cyanobacteria, septal junctions enable molecular exchange between sister cells and are required for cellular differentiation. The structure of septal junctions is poorly understood, and it is unknown whether they are capable of controlling intercellular communication. Here, we resolved the in situ architecture of septal junctions by electron cryotomography of cryo-focused ion beam-milled cyanobacterial filaments. Septal junctions consisted of a tube traversing the septal peptidoglycan. Each tube end comprised a FraD-containing plug, which was covered by a cytoplasmic cap. Fluorescence recovery after photobleaching showed that intercellular communication was blocked upon stress. Gating was accompanied by a reversible conformational change of the septal junction cap. We provide the mechanistic framework for a cell junction that predates eukaryotic gap junctions by a billion years. The conservation of a gated dynamic mechanism across different domains of life emphasizes the importance of controlling molecular exchange in multicellular organisms.


Assuntos
Junções Comunicantes/metabolismo , Anabaena/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Carbonil Cianeto m-Clorofenil Hidrazona/análogos & derivados , Carbonil Cianeto m-Clorofenil Hidrazona/farmacologia , Comunicação Celular/efeitos dos fármacos , Microscopia Crioeletrônica , Junções Comunicantes/química , Junções Comunicantes/ultraestrutura , Proteínas de Membrana/química , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Mutagênese
2.
Annu Rev Microbiol ; 76: 597-618, 2022 09 08.
Artigo em Inglês | MEDLINE | ID: mdl-35671534

RESUMO

Heterocyst differentiation that occurs in some filamentous cyanobacteria, such as Anabaena sp. PCC 7120, provides a unique model for prokaryotic developmental biology. Heterocyst cells are formed in response to combined-nitrogen deprivation and possess a microoxic environment suitable for nitrogen fixation following extensive morphological and physiological reorganization. A filament of Anabaena is a true multicellular organism, as nitrogen and carbon sources are exchanged among different cells and cell types through septal junctions to ensure filament growth. Because heterocysts are terminally differentiated cells and unable to divide, their activity is an altruistic behavior dedicated to providing fixed nitrogen for neighboring vegetative cells. Heterocyst development is also a process of one-dimensional pattern formation, as heterocysts are semiregularly intercalated among vegetative cells. Morphogens form gradients along the filament and interact with each other in a fashion that fits well into the Turing model, a mathematical framework to explain biological pattern formation.


Assuntos
Anabaena , Cianobactérias , Anabaena/metabolismo , Proteínas de Bactérias/metabolismo , Cianobactérias/metabolismo , Regulação Bacteriana da Expressão Gênica , Nitrogênio/metabolismo , Fixação de Nitrogênio
3.
PLoS Biol ; 22(3): e3002546, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38466754

RESUMO

Bacteria have developed fine-tuned responses to cope with potential zinc limitation. The Zur protein is a key player in coordinating this response in most species. Comparative proteomics conducted on the cyanobacterium Anabaena highlighted the more abundant proteins in a zur mutant compared to the wild type. Experimental evidence showed that the exoprotein ZepA mediates zinc uptake. Genomic context of the zepA gene and protein structure prediction provided additional insights on the regulation and putative function of ZepA homologs. Phylogenetic analysis suggests that ZepA represents a primordial system for zinc acquisition that has been conserved for billions of years in a handful of species from distant bacterial lineages. Furthermore, these results show that Zur may have been one of the first regulators of the FUR family to evolve, consistent with the scarcity of zinc in the ecosystems of the Archean eon.


Assuntos
Anabaena , Zinco , Zinco/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Ecossistema , Filogenia , Anabaena/genética , Anabaena/metabolismo , Regulação Bacteriana da Expressão Gênica
4.
Nucleic Acids Res ; 52(1): 404-419, 2024 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-38000383

RESUMO

The bacterial ribonuclease RNase E plays a key role in RNA metabolism. Yet, with a large substrate spectrum and poor substrate specificity, its activity must be well controlled under different conditions. Only a few regulators of RNase E are known, limiting our understanding on posttranscriptional regulatory mechanisms in bacteria. Here we show that, RebA, a protein universally present in cyanobacteria, interacts with RNase E in the cyanobacterium Anabaena PCC 7120. Distinct from those known regulators of RNase E, RebA interacts with the catalytic region of RNase E, and suppresses the cleavage activities of RNase E for all tested substrates. Consistent with the inhibitory function of RebA on RNase E, depletion of RNase E and overproduction of RebA caused formation of elongated cells, whereas the absence of RebA and overproduction of RNase E resulted in a shorter-cell phenotype. We further showed that the morphological changes caused by altered levels of RNase E or RebA are dependent on their physical interaction. The action of RebA represents a new mechanism, potentially conserved in cyanobacteria, for RNase E regulation. Our findings provide insights into the regulation and the function of RNase E, and demonstrate the importance of balanced RNA metabolism in bacteria.


Assuntos
Anabaena , Endorribonucleases , Anabaena/genética , Cianobactérias/genética , Cianobactérias/metabolismo , Endorribonucleases/genética , Endorribonucleases/metabolismo , RNA , RNA Bacteriano/genética , RNA Bacteriano/metabolismo
5.
Proc Natl Acad Sci U S A ; 119(36): e2207963119, 2022 09 06.
Artigo em Inglês | MEDLINE | ID: mdl-36037363

RESUMO

The filamentous, multicellular cyanobacterium Anabaena sp. PCC 7120 (Anabaena) is a prokaryotic model for the study of cell differentiation and cell-cell interactions. Upon combined-nitrogen deprivation, Anabaena forms a particular cell type, heterocyst, for aerobic nitrogen fixation. Heterocysts are semiregularly spaced among vegetative cells. Heterocyst differentiation is coupled to cell division, but the underlying mechanism remains unclear. This mechanism could be mediated by the putative protease HetF, which is a divisome component and is necessary for heterocyst differentiation. In this study, by suppressor screening, we identified PatU3, as a negative regulator acting downstream of HetF for cell division and heterocyst development. The inactivation of patU3 restored the capacity of cell division and heterocyst differentiation in the ΔhetF mutant, and overexpression of patU3 inhibited both processes in the wild-type background. We demonstrated that PatU3 was a specific substrate of the protease activity of HetF. Consequently, PatU3 accumulated in the hetF-deficient mutant, which was responsible for the resultant mutant phenotype. The cleavage site of PatU3 by HetF was mapped after the Arg117 residue, whose mutation made PatU3 resistant to HetF processing, and mimicked the effect of hetF deletion. Our results provided evidence that HetF regulated cell division and heterocyst differentiation by controlling the inhibitory effects of PatU3. This proteolytic pathway constituted a mechanism for the coordination between cell division and differentiation in a prokaryotic model used for studies on developmental biology and multicellularity.


Assuntos
Anabaena , Proteínas de Bactérias , Divisão Celular , Proteólise , Anabaena/citologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica
6.
Mol Microbiol ; 119(4): 492-504, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36756754

RESUMO

Under nitrogen-limiting conditions, the filamentous cyanobacterium Nostoc PCC7120 differentiates nitrogen-fixing heterocysts at semi-regular intervals along filaments generating a periodic pattern of two distinct cell types. Heterocysts are micro-oxic cells that host the oxygen-sensitive nitrogenase allowing two antagonistic activities to take place simultaneously. Although several factors required to control the differentiation process are known, the molecular mechanisms engaged have only been elucidated for a few of them. The patB (cnfR) gene has been shown to be essential for heterocyst formation and nitrogen fixation in this cyanobacterium, but its function remains to be clarified. Here, we show that PatB acts as a direct transcriptional regulator of genes required for nitrogenase production and activity. The DNA-binding activity of PatB does not depend on micro-oxia as it interacts with its target promoters under aerobic conditions both in vitro and in vivo. The absence of the DNA-binding domain of PatB can be rescued in the heterocyst but not in the vegetative cell. Furthermore, the putative ferredoxin domain of PatB is not essential to its interaction with DNA. The patB gene is widely conserved in cyanobacterial genomes and its function can be pleiotropic since it is not limited to nitrogen fixation control.


Assuntos
Anabaena , Nostoc , Proteínas de Bactérias/metabolismo , Nostoc/genética , Nostoc/metabolismo , Fixação de Nitrogênio/genética , Nitrogenase/metabolismo , Nitrogênio/metabolismo , Regulação Bacteriana da Expressão Gênica/genética , Anabaena/metabolismo
7.
Mol Microbiol ; 120(5): 740-753, 2023 11.
Artigo em Inglês | MEDLINE | ID: mdl-37804047

RESUMO

The filamentous cyanobacterium Anabaena sp. PCC 7120 is able to form heterocysts for nitrogen fixation. Heterocyst differentiation is initiated by combined-nitrogen deprivation, followed by the commitment step during which the developmental process becomes irreversible. Mature heterocysts are terminally differentiated cells unable to divide, and cell division is required for heterocyst differentiation. Previously, we have shown that the HetF protease regulates cell division and heterocyst differentiation by cleaving PatU3, which is an inhibitor for both events. When hetF is required during the developmental program remains unknown. Here, by controlling the timing of hetF expression during heterocyst differentiation, we provide evidence that hetF is required just before the beginning of heterocyst morphogenesis. Consistent with this finding, transcriptome data show that most of the genes known to be involved in the early step (such as hetR and ntcA) or the commitment step (such as hetP and hetZ) of heterocyst development could be expressed in the ΔhetF mutant. In contrast, most of the genes involved in heterocyst morphogenesis and nitrogen fixation remain repressed in the mutant. These results indicated that in the absence of hetF, heterocyst differentiation is able to be initiated and proceeds to the stage just before heterocyst envelope formation.


Assuntos
Anabaena , Cianobactérias , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica/genética , Cianobactérias/metabolismo , Diferenciação Celular
8.
Plant Cell Physiol ; 65(6): 975-985, 2024 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-38147500

RESUMO

DesC1 and DesC2, which are fatty acid desaturases found in cyanobacteria, are responsible for introducing a double bond at the Δ9 position of fatty-acyl chains, which are subsequently esterified to the sn-1 and sn-2 positions of the glycerol moiety, respectively. However, since the discovery of these two desaturases in the Antarctic cyanobacterium Nostoc sp. SO-36, no further research has been reported. This study presents a comprehensive characterization of DesC1 and DesC2 through targeted mutagenesis and transformation using two cyanobacteria strains: Anabaena sp. PCC 7120, comprising both desaturases, and Synechocystis sp. PCC 6803, containing a single Δ9 desaturase (hereafter referred to as DesCs) sharing similarity with DesC1 in amino acid sequence. The results suggested that both DesC1 and DesC2 were essential in Anabaena sp. PCC 7120 and that DesC1, but not DesC2, complemented DesCs in Synechocystis sp. PCC 6803. In addition, DesC2 from Anabaena sp. PCC 7120 desaturated fatty acids esterified to the sn-2 position of the glycerol moiety in Synechocystis sp. PCC 6803.


Assuntos
Anabaena , Proteínas de Bactérias , Ácidos Graxos Dessaturases , Synechocystis , Ácidos Graxos Dessaturases/metabolismo , Ácidos Graxos Dessaturases/genética , Synechocystis/enzimologia , Synechocystis/genética , Anabaena/enzimologia , Anabaena/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Ácidos Graxos/metabolismo , Cianobactérias/enzimologia , Cianobactérias/genética , Sequência de Aminoácidos
9.
Arch Microbiol ; 206(3): 105, 2024 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-38363385

RESUMO

Colonization of the cyanobacteria in the Bishnupur terracotta temples, one of the heritage sites of West Bengal, India is in an alarming state of deterioration now. Among the cyanobacteria Anabaena sp. (VBCCA 052002) has been isolated from most of the crust samples of terracotta monuments of Bishnupur. The identification was done using micromorphological characters and confirmed by 16S rRNA gene sequencing. The isolated strain produces enormous exopolysaccharides, which are extracted, hydrolyzed, and analyzed by HPLC. We have studied desiccation tolerance in this cyanobacterium and found biosynthesis of trehalose with an increase in durations of desiccation. The in vitro experiment shows that Chlorophyll-a and carotenoid content increase with fourteen days of desiccation, and cellular carbohydrates increase continuously. However, cellular protein decreases with desiccation. To gain insights into the survival strategies and biodeterioration mechanisms of Anabaena sp. in the desiccated conditions of the Bishnupur monuments, the present study focuses on the physiological aspects of the cyanobacteria under controlled in vitro conditions. Our study indicates that in desiccation conditions, trehalose biosynthesis takes place in Anabaena sp. As a result of the excessive sugar and polysaccharide produced, it adheres to the surface of the terracotta structure. The continuous contraction and expansion of these polysaccharides contribute to the biodeterioration of these monuments.


Assuntos
Anabaena , Dessecação , RNA Ribossômico 16S/genética , Trealose/metabolismo , Anabaena/metabolismo , Polissacarídeos/metabolismo
10.
Environ Res ; 249: 118310, 2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38331154

RESUMO

Organophosphorus (OP) insecticides are widely used for on-field pest control, constituting about 38% of global pesticide consumption. Insecticide tolerance has been recorded in microorganisms isolated from the contaminated soil. However, the cross-tolerance of laboratory-enriched cultures remains poorly understood. A chlorpyrifos tolerant (T) strain of Anabaena sp. PCC 7119 was developed through continuous enrichment of the wild strain (W). The cross-tolerance of the T strain to the OP insecticide dimethoate was assessed by measuring photosynthetic performance, key enzyme activities and degradation potential. The presence of dimethoate led to a significant reduction in the growth and pigment content of the W strain. In contrast, the T strain demonstrated improved growth and metabolic performance. Chl a and carotenoids were degraded faster than phycobiliproteins in both strains. The T strain exhibited superior photosynthetic performance, metabolic efficiency and photosystem functions, than of W strain, at both the tested dimethoate concentrations (100 and 200 µM). The treated T strain had more or less a normal OJIP fluorescence transient and bioenergetic functions, while the W strain showed a greater fluorescence rise at ≤ 300 µs indicating the inhibition of electron donation to PS II, and at 2 ms due to reduced electron release beyond QA. The T strain had significantly higher levels of esterase and phosphatases, further enhanced by insecticide treatment. Dimethoate degradation efficiency of the T strain was significantly higher than of the W strain. T strain also removed chlorpyrifos more efficiently than W strain at both the tested concentrations. The BCFs of both chlorpyrifos and dimethoate were lower in the T strain compared to the W strain. These findings suggest that the enriched strain exhibits promising results in withstanding dimethoate toxicity and could be explored for its potential as a bioremediating organism for OP degradation.


Assuntos
Anabaena , Clorpirifos , Dimetoato , Inseticidas , Clorpirifos/toxicidade , Dimetoato/toxicidade , Anabaena/efeitos dos fármacos , Inseticidas/toxicidade , Fotossíntese/efeitos dos fármacos
11.
Mol Cell Proteomics ; 21(4): 100224, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35288331

RESUMO

The filamentous cyanobacterium Anabaena sp. PCC 7120 can differentiate into heterocysts to fix atmospheric nitrogen. During cell differentiation, cellular morphology and gene expression undergo a series of significant changes. To uncover the mechanisms responsible for these alterations, we built protein-protein interaction (PPI) networks for these two cell types by cofractionation coupled with mass spectrometry. We predicted 280 and 215 protein complexes, with 6322 and 2791 high-confidence PPIs in vegetative cells and heterocysts, respectively. Most of the proteins in both types of cells presented similar elution profiles, whereas the elution peaks of 438 proteins showed significant changes. We observed that some well-known complexes recruited new members in heterocysts, such as ribosomes, diflavin flavoprotein, and cytochrome c oxidase. Photosynthetic complexes, including photosystem I, photosystem II, and phycobilisome, remained in both vegetative cells and heterocysts for electron transfer and energy generation. Besides that, PPI data also reveal new functions of proteins. For example, the hypothetical protein Alr4359 was found to interact with FraH and Alr4119 in heterocysts and was located on heterocyst poles, thereby influencing the diazotrophic growth of filaments. The overexpression of Alr4359 suspended heterocyst formation and altered the pigment composition and filament length. This work demonstrates the differences in protein assemblies and provides insight into physiological regulation during cell differentiation.


Assuntos
Anabaena , Regulação Bacteriana da Expressão Gênica , Anabaena/genética , Anabaena/metabolismo , Proteínas de Bactérias/metabolismo , Biologia , Diferenciação Celular
12.
Biochem J ; 480(1): 87-104, 2023 01 13.
Artigo em Inglês | MEDLINE | ID: mdl-36594794

RESUMO

Thioredoxins (Trxs) are ubiquitous proteins that play vital roles in several physiological processes. Alr2205, a thioredoxin-like protein from Anabaena PCC 7120, was found to be evolutionarily closer to the Trx-domain of the NADPH-Thioredoxin Reductase C than the other thioredoxins. The Alr2205 protein showed disulfide reductase activity despite the presence a non-canonical active site motif 'CPSC'. Alr2205 not only physically interacted with, but also acted as a physiological reductant of Alr4641 (the typical 2-Cys-Peroxiredoxin from Anabaena), supporting its peroxidase function. Structurally, Alr2205 was a monomeric protein that formed an intramolecular disulfide bond between the two active site cysteines (Cys-38 and Cys-41). However, the Alr2205C41S protein, wherein the resolving cysteine was mutated to serine, was capable of forming intermolecular disulfide bond and exist as a dimer when treated with H2O2. Overproduction of Alr2205 in E. coli protected cells from heavy metals, but not oxidative stress. To delve into its physiological role, Alr2205/Alr2205C41S was overexpressed in Anabaena, and the ability of the corresponding strains (An2205+ or An2205C41S+) to withstand environmental stresses was assessed. An2205+ showed higher resistance to H2O2 than An2205C41S+, indicating that the disulfide reductase function of this protein was critical to protect cells from this peroxide. Although, An2205+ did not show increased capability to withstand cadmium stress, An2205C41S+ was more susceptible to this heavy metal. This is the first study that provides a vital understanding into the function of atypical thioredoxins in countering the toxic effects of heavy metals/H2O2 in prokaryotes.


Assuntos
Anabaena , Cianobactérias , Peroxirredoxinas/genética , Peroxirredoxinas/metabolismo , Peróxido de Hidrogênio/metabolismo , Cisteína/genética , Cisteína/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Oxirredução , Proteínas de Bactérias/metabolismo , Anabaena/genética , Anabaena/metabolismo , Cianobactérias/metabolismo , Tiorredoxinas/química , Dissulfetos/metabolismo , Tiorredoxina Dissulfeto Redutase/genética , Tiorredoxina Dissulfeto Redutase/metabolismo
13.
J Plant Res ; 137(3): 521-543, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38460108

RESUMO

The present study examined the regulatory mechanism of hydrogen sulfide (H2S) and nitric oxide (NO) in nickel (Ni) stressed cyanobacteria viz., Nostoc muscorum and Anabaena sp. by analyzing growth, photosynthetic pigments, biochemical components (protein and carbohydrate), exopolysaccharides (EPS), inorganic nitrogen content, and activity of enzymes comprised in nitrogen metabolism and Ni accumulation. The 1 µM Ni substantially diminished growth by 18% and 22% in N. muscorum and Anabaena sp. respectively, along with declining the pigment contents (Chl a/Car ratio and phycobiliproteins), and biochemical components. It also exerted negative impacts on inorganic uptake of nitrate and nitrite contents; nitrate reductase and nitrite reductase; and ammonium assimilating enzymes (glutamine synthetase, glutamate synthase, and glutamate dehydrogenase exhibited a reverse trend) activities. Nonetheless, the adverse impact of Ni can be mitigated through the exogenous supplementation of NaHS [sodium hydrosulfide (8 µM); H2S donor] and SNP [sodium nitroprusside (10 µM); NO donor] which showed substantial improvement on growth, pigments, nitrogen metabolism, and EPS layer and noticeably occurred as a consequence of a substantial reduction in Ni accumulation content which minimized the toxicity effects. The accumulation of Ni on both the cyanobacterial cell surface (EPS layer) are confirmed by the SEM-EDX analysis. Further, the addition of NO scavenger (PTIO; 20 µM) and inhibitor of NO (L-NAME; 100 µM); and H2S scavenger (HT; 20 µM) and H2S inhibitor (PAG; 50 µM) reversed the positive responses of H2S and NO and damages were more prominent under Ni stress thereby, suggesting the downstream signaling of H2S on NO-mediated alleviation. Thus, this study concludes the crosstalk mechanism of H2S and NO in the mitigation of Ni-induced toxicity in rice field cyanobacteria.


Assuntos
Sulfeto de Hidrogênio , Níquel , Óxido Nítrico , Nitrogênio , Oryza , Óxido Nítrico/metabolismo , Níquel/metabolismo , Sulfeto de Hidrogênio/metabolismo , Nitrogênio/metabolismo , Oryza/metabolismo , Oryza/efeitos dos fármacos , Oryza/crescimento & desenvolvimento , Nostoc muscorum/metabolismo , Polissacarídeos Bacterianos/metabolismo , Anabaena/metabolismo , Anabaena/efeitos dos fármacos , Anabaena/crescimento & desenvolvimento , Estresse Fisiológico , Nitroprussiato/farmacologia
14.
Water Sci Technol ; 89(5): 1142-1154, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38483490

RESUMO

The efficiency of Anabaena sp. was analyzed for the phytoremediation of wastewater loaded with organic matter and heavy metals like chromium. Simulated wastewater was contaminated with chromium. A side-stream membrane bioreactor was used for the treatment of wastewater. A feed tank of 20 L capacity was used with a stirring arrangement. A ceramic microfiltration membrane composed of clay and alumina was obtained from Johnson & Johnson. The removal efficiency of chemical oxygen demand, biochemical oxygen demand, and chromium was evaluated. The process was used for algae harvesting and wastewater treatment. About 92% of chemical oxygen demand (COD), 98% chromium, and oil and grease were completely removed. Membrane fouling was explained by the pore blocking and cake resistance model. Stress in algal cells was determined from the superoxide dismutase (SOD) and catalase (CAT) analysis. The lipid content of algal cells was measured.


Assuntos
Anabaena , Cromo , Águas Residuárias , Biodegradação Ambiental , Reatores Biológicos
15.
Environ Microbiol ; 25(11): 2142-2162, 2023 11.
Artigo em Inglês | MEDLINE | ID: mdl-37315963

RESUMO

Zinc is required for the activity of many enzymes and plays an essential role in gene regulation and redox homeostasis. In Anabaena (Nostoc) sp. PCC7120, the genes involved in zinc uptake and transport are controlled by the metalloregulator Zur (FurB). Comparative transcriptomics of a zur mutant (Δzur) with the parent strain unveiled unexpected links between zinc homeostasis and other metabolic pathways. A notable increase in the transcription of numerous desiccation tolerance-related genes, including genes involved in the synthesis of trehalose and the transference of saccharide moieties, among many others, was detected. Biofilm formation analysis under static conditions revealed a reduced capacity of Δzur filaments to form biofilms compared to the parent strain, and such capacity was enhanced when Zur was overexpressed. Furthermore, microscopy analysis revealed that zur expression is required for the correct formation of the envelope polysaccharide layer in the heterocyst, as Δzur cells showed reduced staining with alcian blue compared to Anabaena sp. PCC7120. We suggest that Zur is an important regulator of the enzymes involved in the synthesis and transport of the envelope polysaccharide layer, influencing heterocyst development and biofilm formation, both relevant processes for cell division and interaction with substrates in its ecological niche.


Assuntos
Anabaena , Metais , Metais/metabolismo , Zinco/metabolismo , Homeostase , Polissacarídeos/metabolismo , Anabaena/genética , Anabaena/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica
16.
Biochem Biophys Res Commun ; 639: 134-141, 2023 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-36493556

RESUMO

In cyanobacteria and algae (but not plants), flavodoxin (Fld) replaces ferredoxin (Fd) under stress conditions to transfer electrons from photosystem I (PSI) to ferredoxin-NADP+ reductase (FNR) during photosynthesis. Fld constitutes a small electron carrier noncovalently bound to flavin mononucleotide (FMN), and also an ideal model for revealing the protein/flavin-binding mechanism because of its relative simplicity compared to other flavoproteins. Here, we report two crystal structures of apo-Fld from Synechococcus sp. PCC 7942, one dimeric structure of 2.09 Å and one monomeric structure of 1.84 Å resolution. Analytical ultracentrifugation showed that in solution, apo-Fld exists both as monomers and dimers. Our dimer structure contains two ligand-binding pockets separated by a distance of 45 Å, much longer than the previous structures of FMN-bound dimers. These results suggested a potential dimer-monomer transition mechanism of cyanobacterial apo-Fld. We further propose that the dimer represents the "standby" state to stabilize itself, while the monomer constitutes the "ready" state to bind FMN. Furthermore, we generated a new docking model of cyanobacterial Fld-FNR complex based on the recently reported cryo-EM structures, and mapped the special interactions between Fld and FNR in detail.


Assuntos
Anabaena , Cianobactérias , Flavodoxina/química , Flavodoxina/metabolismo , Ferredoxinas/metabolismo , Anabaena/metabolismo , Flavoproteínas , Ferredoxina-NADP Redutase/química , Cianobactérias/metabolismo , Oxirredução
17.
Biochem Biophys Res Commun ; 682: 316-324, 2023 11 19.
Artigo em Inglês | MEDLINE | ID: mdl-37837752

RESUMO

Muscle cell cultivation, specifically the culture of artificial meat from livestock-derived cells in serum-free media is an emerging technology and has attracted much attention. However, till now, the high cost of production and environmental load have been significant deterrents. This study aims to provide an alternate growth-promoting substance that is free from animal derivatives and lowers nitrogen pollution. We have extracted water-soluble compounds from the filamentous nitrogen-fixing cyanobacteria Anabaena sp. PCC 7120 by the ultrasonication method. The heat-inactivated and molecular weight separation experiments were conducted to identify the bioactive compound present in the extract. Finally, the compounds soluble in water (CW) containing the water-soluble pigment protein, phycocyanin as a bioactive compound, was added as a growth supplement to cultivate muscle cells such as C2C12 muscle cells and quail muscle clone 7 (QM7) cells to analyze the effectiveness of the extract. The results indicated that CW had a positive role in muscle cell proliferation. A three-dimensional (3-D) cell-dense structure was fabricated by culturing QM7 cells using the extract. Furthermore, the nitrogen-fixing cyanobacterial extract has vast potential for cultured meat production without animal sera in the near future.


Assuntos
Anabaena , Cianobactérias , Nitrogênio/metabolismo , Proteínas de Bactérias/metabolismo , Cianobactérias/metabolismo , Anabaena/metabolismo , Músculos/metabolismo , Proliferação de Células , Regulação Bacteriana da Expressão Gênica
18.
New Phytol ; 237(1): 126-139, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36128660

RESUMO

The model heterocyst-forming filamentous cyanobacterium Anabaena sp. PCC 7120 (Anabaena) is a typical example of a multicellular organism capable of simultaneously performing oxygenic photosynthesis in vegetative cells and O2 -sensitive N2 -fixation inside heterocysts. The flavodiiron proteins have been shown to participate in photoprotection of photosynthesis by driving excess electrons to O2 (a Mehler-like reaction). Here, we performed a phenotypic and biophysical characterization of Anabaena mutants impaired in vegetative-specific Flv1A and Flv3A in order to address their physiological relevance in the bioenergetic processes occurring in diazotrophic Anabaena under variable CO2 conditions. We demonstrate that both Flv1A and Flv3A are required for proper induction of the Mehler-like reaction upon a sudden increase in light intensity, which is likely important for the activation of carbon-concentrating mechanisms and CO2 fixation. Under ambient CO2 diazotrophic conditions, Flv3A is responsible for moderate O2 photoreduction, independently of Flv1A, but only in the presence of Flv2 and Flv4. Strikingly, the lack of Flv3A resulted in strong downregulation of the heterocyst-specific uptake hydrogenase, which led to enhanced H2 photoproduction under both oxic and micro-oxic conditions. These results reveal a novel regulatory network between the Mehler-like reaction and the diazotrophic metabolism, which is of great interest for future biotechnological applications.


Assuntos
Anabaena , Dióxido de Carbono , Dióxido de Carbono/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Anabaena/genética , Anabaena/metabolismo , Oxigênio/metabolismo , Fotossíntese/fisiologia
19.
PLoS Comput Biol ; 18(8): e1010359, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35969646

RESUMO

The Anabaena genus is a model organism of filamentous cyanobacteria whose vegetative cells can differentiate under nitrogen-limited conditions into a type of cell called heterocyst. These heterocysts lose the possibility to divide and are necessary for the colony because they can fix and share environmental nitrogen. In order to distribute the nitrogen efficiently, heterocysts are arranged to form a quasi-regular pattern whose features are maintained as the filament grows. Recent efforts have allowed advances in the understanding of the interactions and genetic mechanisms underlying this dynamic pattern. However, the main role of the patA and hetF genes are yet to be clarified; in particular, the patA mutant forms heterocysts almost exclusively in the terminal cells of the filament. In this work, we investigate the function of these genes and provide a theoretical model that explains how they interact within the broader genetic network, reproducing their knock-out phenotypes in several genetic backgrounds, including a nearly uniform concentration of HetR along the filament for the patA mutant. Our results suggest a role of hetF and patA in a post-transcriptional modification of HetR which is essential for its regulatory function. In addition, the existence of molecular leakage out of the filament in its boundary cells is enough to explain the preferential appearance of terminal heterocysts, without any need for a distinct regulatory pathway.


Assuntos
Anabaena , Regulação Bacteriana da Expressão Gênica , Anabaena/genética , Anabaena/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica/genética , Redes Reguladoras de Genes , Nitrogênio/metabolismo
20.
Fish Shellfish Immunol ; 139: 108865, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37277048

RESUMO

Shrimp are especially susceptible to the White Spot Syndrome Virus (WSSV). Oral administration of the WSSV envelop protein VP28 is a promising approach to protect shrimp against WSSV. In this study, Macrobrachium nipponense (M. nipponense) were fed for 7 days with food supplemented with Anabaena sp. PCC 7120 (Ana7120) expressing VP28 and then challenged with WSSV. The survival rates of M. nipponense in three groups, including control, WSSV-challenged, and VP28-vaccinated, were subsequently determined. We also determined the WSSV content of different tissues and the tissue morphology in the absence of and after viral challenge. The survival rate of the positive control group (no vaccination and challenge, 10%) and empty vector group (fed with Ana7120 pRL-489 algae and challenged, 13.3%) was much lower than the survival rate of M. nipponense in wild type group (fed with Ana7120 and challenged, 18.9%), immunity group 1 (fed with 3.33% Ana7120 pRL-489-vp28 and challenged, 45.6%) or immunity group 2 (fed with 6.66% Ana7120 pRL-489-vp28 and challenged, 62.2%). RT-qPCR showed that WSSV content of the gill, hepatopancreas and muscle of immunity groups 1 and 2 were substantially lower than the positive control. Microscopic examination revealed that WSSV-challenged positive control exhibited large number of cell rupture, necrosis, nuclear exfoliation in gills and hepatopancreatic tissues. The gill and hepatopancreas of immunity group 1 showed partial symptoms of infection, yet the tissue was visibly healthier than that of the positive control group. No symptoms were visible in the gills and hepatopancreatic tissue of immunity group 2. The results demonstrate that the probability of M. nipponense infected by WSSV can be diminished by oral administration of cyanobacteria-expressed VP28. Such an approach could improve the disease resistance and delay the death of M. nipponense in the commercial production of this shrimp.


Assuntos
Anabaena , Palaemonidae , Penaeidae , Vírus da Síndrome da Mancha Branca 1 , Animais , Vírus da Síndrome da Mancha Branca 1/fisiologia , Microscopia , Proteínas do Envelope Viral
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