Comparative proteomics of oxidative stress response in three cyanobacterial strains native to Indian paddy fields

Panda, Bandita; Basu, Bhakti; Rajaram, Hema; Apte, Shree Kumar

doi:10.1016/j.jprot.2015.05.020

Cited by 14 publications

(5 citation statements)

References 21 publications

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“…punctiforme homologues of these were: Npun_R0971, a HHE cation-binding protein; Npun_F3786, a signal transduction histidine kinase homologue containing four predicted transmembrane domains; Npun_F3789, a high light inducible protein; and Npun_R4582, a manganese-containing catalase homologue [48]. No similar parallels between gene expression in the 2 g strain and similar proteomic experiments testing for responses to oxidative stress were apparent [50]. As butachlor is a hydrophobic compound, these gene responses point to parallels between exposure to this herbicide and internal production of alkanes that may be unique to stress associated with hydrophobic compounds.…”

Section: Resultsmentioning

confidence: 99%

Transcriptomic analysis of cyanobacterial alkane overproduction reveals stress-related genes and inhibitors of lipid droplet formation

et al. 2020

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The cyanobacterium Nostoc punctiforme can form lipid droplets (LDs), internal inclusions containing triacylglycerols, carotenoids and alkanes. LDs are enriched for a 17 carbon-long alkane in N. punctiforme , and it has been shown that the overexpression of the aar and ado genes results in increased LD and alkane production. To identify transcriptional adaptations associated with increased alkane production, we performed comparative transcriptomic analysis of an alkane overproduction strain. RNA-seq data identified a large number of highly upregulated genes in the overproduction strain, including genes potentially involved in rRNA processing, mycosporine-glycine production and synthesis of non-ribosomal peptides, including nostopeptolide A. Other genes encoding helical carotenoid proteins, stress-induced proteins and those for microviridin synthesis were also upregulated. Construction of N. punctiforme strains with several upregulated genes or operons on multi-copy plasmids resulted in reduced alkane accumulation, indicating possible negative regulators of alkane production. A strain containing four genes for microviridin biosynthesis completely lost the ability to synthesize LDs. This strain exhibited wild-type growth and lag phase recovery under standard conditions, and slightly faster growth under high light. The transcriptional changes associated with increased alkane production identified in this work will provide the basis for future experiments designed to use cyanobacteria as a production platform for biofuel or high-value hydrophobic products.

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Section: Resultsmentioning

confidence: 99%

Transcriptomic analysis of cyanobacterial alkane overproduction reveals stress-related genes and inhibitors of lipid droplet formation

et al. 2020

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“…The genome of Anabaena encodes 6132 predicted proteins. Previous studies mainly focused on proteome dynamics in various growth conditions, using 2D gel‐based approaches identifying < 100 differentially regulated proteins . The most comprehensive proteome analysis was carried out by Ow et al., identifying 506 proteins in Anabaena enriched heterocysts preparations …”

Section: Resultsmentioning

confidence: 99%

Proteome Analysis of Enriched Heterocysts from Two Hydrogenase Mutants from Anabaena sp. PCC 7120

et al. 2019

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Cyanobacteria are oxygenic photosynthetic prokaryotes and play a crucial role in the Earth's carbon and nitrogen cycles. The photoautotrophic cyanobacterium Anabaena sp. PCC 7120 has the ability to fix atmospheric nitrogen in heterocysts and produce hydrogen as a byproduct through a nitrogenase. In order to improve hydrogen production, mutants from Anabaena sp. PCC 7120 are constructed by inactivation of the uptake hydrogenase (ΔhupL) and the bidirectional hydrogenase (ΔhoxH) in previous studies. Here the proteomic differences of enriched heterocysts between these mutants cultured in N2‐fixing conditions are investigated. Using a label‐free quantitative proteomics approach, a total of 2728 proteins are identified and it is found that 79 proteins are differentially expressed in the ΔhupL and 117 proteins in the ΔhoxH variant. The results provide for the first time comprehensive information on proteome regulation of the uptake hydrogenase and the bidirectional hydrogenase, as well as systematic data on the hydrogen related metabolism in Anabaena sp. PCC 7120.

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“…To analyze the transcript level of alr0765, total RNA was extracted using RNASure mini kit (Nucleo-pore) from 100 mL culture (OD 750 nm 0.6) of Anabaena PCC7120 treated with LC 50 dose of arsenic (40 mM for 24 h) [ 24 ], salt (100 mM for 24 h) [ 25 ], heat (42°C for 1 h) [ 26 ], UV-B (12.9 mWcm -2 for 30 min) [ 27 ], drought (30°C for 24 h) [ 28 ], butachlor (32µM for 24 h) [ 29 ], cadmium (10 µM for 24 h) [ 30 ] and methyl viologen (2 μM for 6 h) [ 31 ] and without treatment (control). cDNA from RNA was prepared using the iScript cDNA synthesis kit (BioRad).…”

Section: Methodsmentioning

confidence: 99%

Functional Characterization of Alr0765, A Hypothetical Protein from Anabaena PCC 7120 Involved in Cellular Energy Status Sensing, Iron Acquisition and Abiotic Stress Management in E. coli Using Molecular, Biochemical and Computational Approaches

Chatterjee

Singh

Rai

et al. 2020

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Background: Cyanobacteria are excellent model to understand the basic metabolic processes taking place in response to abiotic stress. The present study involves characterization of a hypothetical protein Alr0765 of Anabaena PCC7120 comprising CBS-CP12 domain and deciphering its role in abiotic stress tolerance. Methods: Molecular cloning, heterologous expression and protein purification using affinity chromatography was performed to obtain native purified protein Alr0765. Energy sensing property of Alr0765 was inferred from its binding affinity with different ligand molecules as analyzed by FTIR and TNP-ATP binding assay. AAS and real time-PCR were applied to evaluate the iron acquisition property and cyclic voltammetry was employed to check redox sensitivity of the target protein. Transcript level under different abiotic stresses as well as spot assay, CFU count, ROS level and cellular H2O2 level were used to show potential role of Alr0765 in abiotic stress tolerance. In-silico analysis of Alr0765 included molecular function probability analysis, multiple sequence analysis, protein domain and motif finding, secondary structure analysis, protein ligand interaction, homologous modeling, model refinement and verification and molecular docking was performed with COFACTOR, PROMALS-3D, InterProScan, MEME, TheaDomEx, COACH, Swiss modeller, Modrefiner, PROCHECK, ERRAT, MolProbity, ProSA, TM-align, and Discovery studio respectively. Results: Transcript levels of alr0765 significantly increased by 20, 13, 15, 14.8, 12, 7, 6 and 2.5 fold when Anabaena PCC7120 treated with LC50 dose of heat, arsenic, cadmium, butachlor, salt, mannitol (drought), UV-B, and methyl viologen respectively, with respect to control (untreated). Heterologous expression resulted in 23KDa protein observed on the SDS-PAGE. Immunoblotting and MALDI-TOF-MS/MS followed by MASCOT search analysis confirmed the identity of the protein and ESI/MS revealed the purified protein was a dimer. Binding possibility of Alr0765 with ATP was observed with almost 6-fold increment in relative fluorescence during TNP-ATP binding assay with a ƛ max of 538 nm. FTIR spectra revealed modification in protein confirmation upon binding of Alr0765 with ATP, ADP, AMP and NADH. A 10-fold higher accumulation of iron was observed in digests of E. coli with recombinant vector after induction as compared to control affirms the iron acquisition property of protein. Moreover, generation of redox potential of 146 mV by Alr0765 suggested its probable role in maintaining redox status of the cell under environmental constraints. As per CFU count recombinant E. coli BL21 cells showed about 14.7, 7.3, 6.9, 1.9, 3, 4.9 fold higher number of colonies under heat, cadmium (CdCl2), arsenic (Na3AsO4), salt (NaCl), UV-B and drought (mannitol) respectively compared to pET21a harboring E. coli BL21 cells. Deterioration in cellular ROS level and total cellular H2O2 concentration validated stress tolerance ability of Alr0765. In-silico analysis unraveled novel findings and attested experimental findings in determining the role of Alr0765. Conclusion: Alr0765 is a novel CBS-CP12 domain protein that maintains cellular energy level and iron homeostasis provide tolerance against multiple abiotic stresses.

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Comparative proteomics of oxidative stress response in three cyanobacterial strains native to Indian paddy fields

Cited by 14 publications

References 21 publications

Transcriptomic analysis of cyanobacterial alkane overproduction reveals stress-related genes and inhibitors of lipid droplet formation

Transcriptomic analysis of cyanobacterial alkane overproduction reveals stress-related genes and inhibitors of lipid droplet formation

Proteome Analysis of Enriched Heterocysts from Two Hydrogenase Mutants from Anabaena sp. PCC 7120

Functional Characterization of Alr0765, A Hypothetical Protein from Anabaena PCC 7120 Involved in Cellular Energy Status Sensing, Iron Acquisition and Abiotic Stress Management in E. coli Using Molecular, Biochemical and Computational Approaches

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