A5071904322- Photosynthetic Processes and Mechanisms
- Metal-Catalyzed Oxygenation Mechanisms
- Redox biology and oxidative stress
- Metalloenzymes and iron-sulfur proteins
- Plant-Microbe Interactions and Immunity
- Neonatal Health and Biochemistry
- Mitochondrial Function and Pathology
- Porphyrin Metabolism and Disorders
- Antibiotic Resistance in Bacteria
- Trace Elements in Health
- Mycotoxins in Agriculture and Food
- Antifungal resistance and susceptibility
- Biochemical and Molecular Research
- Light effects on plants
- Chemical Analysis and Environmental Impact
- Organ Transplantation Techniques and Outcomes
- Heme Oxygenase-1 and Carbon Monoxide
- Cassava research and cyanide
- Plant pathogens and resistance mechanisms
- CRISPR and Genetic Engineering
- Plant Pathogens and Resistance
- bioluminescence and chemiluminescence research
- Folate and B Vitamins Research
- Antimicrobial Resistance in Staphylococcus
- Advanced Nanomaterials in Catalysis
National University of Rosario
2006-2023
Instituto de Biología Molecular y Celular de Rosario
2006-2023
Consejo Nacional de Investigaciones Científicas y Técnicas
2005-2023
Summary Reactive oxygen species ( ROS ) play fundamental roles in plant responses to pathogen infection, including modulation of cell death processes and defense‐related gene expression. Cell triggered as part the hypersensitive response enhances resistance biotrophic pathogens, but favors virulence necrotrophs. Even though involvement orchestration defense is well established, relative contribution specific subcellular sources against microorganisms with different pathogenesis strategies...
NADP(H) is a central metabolic hub providing reducing equivalents to multiple biosynthetic, regulatory and antioxidative pathways in all living organisms. While biosensors are available determine NADP+ or NADPH levels vivo, no probe exists estimate the redox status, determinant of cell energy availability. We describe herein design characterization genetically-encoded ratiometric biosensor, termed NERNST, able interact with ENADP(H). NERNST consists redox-sensitive green fluorescent protein...
The soxRS regulon protects Escherichia coli cells against superoxide and nitric oxide. Oxidation of the SoxR sensor, a [2Fe-2S]-containing transcriptional regulator, triggers response, but nature cellular signal sensed by is still matter debate. In vivo, sensor maintained in reduced, inactive state activities reductases, which employ NADPH as an electron donor. hypothesis that levels affect deployment response was tested transforming E. with genes encoding enzymes proteins lead to either...
Light significantly influences plant stress responses, with chloroplasts playing a pivotal role as both energy providers and light sensors. They communicate the nucleus through retrograde signals, including secondary metabolites reactive oxygen species (ROS). To investigate contribution of chloroplast redox biochemistry to biotic we studied interactions tobacco leaves expressing alternative electron shuttle flavodoxin virulent nonhost Pseudomonas syringae pathovars under dark conditions....
ABSTRACT Escherichia coli cells from strain fpr , deficient in the soxRS -induced ferredoxin (flavodoxin)-NADP(H) reductase (FPR), display abnormal sensitivity to bactericidal effects of superoxide-generating reagent methyl viologen (MV). Neither bacteriostatic nor inactivation oxidant-sensitive hydrolyases could be detected exposed MV. FPR did not affect MV-driven response, whereas overexpression led enhanced stimulation regulon, with concomitant oxidation NADPH pool. Accumulation a...
The NADP(H)-dependent enzymes glucose-6-phosphate dehydrogenase (G6PDH) and ferredoxin(flavodoxin)-NADP(H) reductase (FPR), encoded by the zwf fpr genes, respectively, are committed members of soxRS regulatory system involved in superoxide resistance Escherichia coli. Exposure E. coli cells to propagator methyl viologen (MV) led rapid accumulation G6PDH, while FPR was induced after a lag period several minutes. Bacteria expressing G6PDH from multicopy plasmid accumulated higher NADPH levels...
Long-chain flavodoxins, ubiquitous electron shuttles containing flavin mononucleotide (FMN) as prosthetic group, play an important protective role against reactive oxygen species (ROS) in various microorganisms. Pseudomonas aeruginosa is opportunistic pathogen which frequently has to face ROS toxicity the environment well within host. We identified a single ORF, hereafter referred fldP (for flavodoxin from P. aeruginosa), displaying highest similarity length, sequence identity and predicted...
Ferredoxin‐NADP + reductases (FNR) participate in cellular defense against oxidative damage. Escherichia coli mutants deficient FNR are abnormally sensitive to methyl viologen and hydrogen peroxide. Tolerance these oxidants was regained by expression of plant FNR, superoxide dismutase, or catalase genes the mutant cells. contribution concerted toxicity under redox‐cycling conditions similar that two major E. dismutases together, as judged phenotypes displayed relevant strains. However, sodA...
Contemporary climate change is characterized by the increased intensity and frequency of environmental stress events such as floods, droughts, heatwaves, which have a debilitating impact on photosynthesis growth, compromising production food, feed, biofuels for an expanding population. The need to increase crop productivity in context global warming has fueled attempts improve several key plant features photosynthetic performance, assimilate partitioning, tolerance stresses. Chloroplast...
Glucose 6-Phosphate Dehydrogenases (G6PDHs) from different sources show varying specificities towards NAD+ and NADP+ as cofactors. However, it is not known to what extent structural determinants of cofactor preference are conserved in the G6PDH family. In this work, molecular simulations, kinetic characterization site-directed mutants phylogenetic analyses were used study basis for strong shown by Escherichia coli. Molecular Dynamics trajectories homology models showed a highly favorable...
We have recently reported the expression in Escherichia coli of an enzymatically competent ferredoxin-NADP+ oxidoreductase from cloned pea genes encoding either mature enzyme or its precursor protein (Ceccarelli, E. A., Viale, A. M., Krapp, R., and Carrillo, N. (1991) J. Biol. Chem. 266, 14283-14287). Processing to form by bacterial protease(s) FAD assembly occurred cytosol. Expression reductase chaperonin-deficient (groE-) mutants resulted partial at permissive growth temperatures (i.e. 30...
The flavoprotein ferredoxin-NADP+ reductase (FNR) catalyzes the final step of photosynthetic electron transport chain, i.e. reduction NADP+ by ferredoxin. A cloned FNR cDNA from a pea library (Newman, B., and Gray, J. (1988) Plant Mol. Biol. 10, 511-520) was used to construct plasmids which express apoenzyme in Escherichia coli. Two recombinant vectors were prepared, one containing sequence corresponding mature enzyme another including, addition, transit peptide that directs chloroplast....
The precursor of the chloroplast flavoprotein ferredoxin-NADP+ reductase from pea was expressed in Escherichia coli as a carboxyl-terminal fusion to glutathione S-transferase. fused protein soluble, and could be purified few steps involving affinity chromatography on glutathione-agarose, cleavage transferase portion by protease Xa, ion exchange DEAE-cellulose. prereductase contained bound FAD but displayed marginally low levels activity. Removal transit peptide limited proteolysis rendered...
Candida dubliniensis is a yeast species closely related to albicans, but in contrast C. limited information available on the virulence factors of this important fungal pathogen. The objective present study was determine if able evoke an adaptive response oxidants. dubliniensis, treated with low concentration either H(2)O(2) or methyl viologen (a superoxide generating agent), mounts that results increased survival against lethal doses both This characterized by induction enzymes known...
The DnaK system is required for the productive folding of pea chloroplast ferredoxin‐NADP + reductase (FNR) expressed in Escherichia coli . formation a mature active enzyme was severely impaired E. dnaK , dnaJ or grpE mutants expressing either cytosolic precursor (preFNR) apoenzyme, and these forms aggregated extensively cells. Coexpression from multicopy plasmid ‐null restored preFNR processing FNR, rendering mature‐sized enzyme. Overexpression GroESL chaperonins failed to prevent...
To achieve reliable detection of methicillin resistance in clinical isolates coagulase-negative staphylococci.Strains (105) were evaluated by normatized antimicrobial susceptibility methods, and for the presence resistance-determining mecA gene, using polymerase chain reaction. Correlation between phenotypic genotypic methods was obtained 87.6% samples. Six strains, classified as methicillin-susceptible assays, revealed indicating that expression probably repressed. Another seven failed to...
The cytosolic precursor of the chloroplast flavoprotein ferredoxin‐NADP + reductase was expressed in Escherichia coli rendering a soluble protein that contained bound FAD and could be imported by isolated chloroplasts. mechanism plastid translocation studied under defined conditions using this recombinant holoprotein intact pea first step import pathway, namely, binding to chloroplasts, saturable at about 2000 molecules/ plastid, showed high‐affinity interaction with dissociation constant K...
Abstract We describe the use of a non‐viral, polyethylenimine‐based vector to transfect rat hepatocytes preserved under hypothermic storage. DNA sequences encoding Escherichia coli β‐galactosidase and pea ferredoxin‐NADP(H) oxidoreductase (FNR), cloned into plasmids pCH110 pKM4 respectively, were used. FNR was detected in liver animals transplanted with transfected cells; no reactivity observed endogenous parenchyma. The expression transgene transient as it detectable up 96 h subsequently...
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Abstract The CRISPR/Cas9 system is widely used for editing genes in various organisms and a very useful tool due to its versatility, simplicity, efficiency. To teach principles post‐graduate students we designed laboratory activity obtain analyze PDS3 mutants Arabidopsis thaliana plants consisting of: 1) Design of guide RNAs using bioinformatics tools; 2) plant transformation (which optional depending on the length course); 3) observation evaluation mutant's phenotypes Phytoene desaturase...