A5009261585- Fungal and yeast genetics research
- Redox biology and oxidative stress
- Polysaccharides and Plant Cell Walls
- RNA and protein synthesis mechanisms
- Endoplasmic Reticulum Stress and Disease
- Trace Elements in Health
- Enzyme Structure and Function
- Antifungal resistance and susceptibility
- Metal-Catalyzed Oxygenation Mechanisms
- Metalloenzymes and iron-sulfur proteins
- Bacterial Genetics and Biotechnology
- DNA Repair Mechanisms
- RNA Research and Splicing
- Glutathione Transferases and Polymorphisms
- RNA modifications and cancer
- Yeasts and Rust Fungi Studies
- Fungal Biology and Applications
- Cardiac Valve Diseases and Treatments
- Nail Diseases and Treatments
- Genomics, phytochemicals, and oxidative stress
- Microbial Metabolites in Food Biotechnology
- CRISPR and Genetic Engineering
- Glycosylation and Glycoproteins Research
- DNA and Nucleic Acid Chemistry
- Cardiac, Anesthesia and Surgical Outcomes
Hospiten Group
2010-2018
Hospital Universitari i Politècnic La Fe
2012-2017
Universitat de Lleida
2007-2016
Biomedical Research Institute of Lleida
2007-2016
Consejo Superior de Investigaciones Científicas
2001-2011
Unidades Centrales Científico-Técnicas
2011
Renfe Operadora (Spain)
2001
Universidad Complutense de Madrid
2001
VIB-UGent Center for Medical Biotechnology
2001
Hospital General Universitario de Alicante Doctor Balmis
1999
A set of Saccharomyces cerevisiae expression vectors has been developed in which transcription is driven by a hybrid tetO-CYC1 promoter through the action tetR-VP16 (tTA) activator. Expression from regulated tetracycline or derivatives. Various modalities and activator are used order to achieve different levels maximal expression. In presence antibiotic growth medium at concentrations that do not affect cell growth, tetO negligible, upon removal induction ratios up 1000-fold observed with...
Yeast cells contain a family of three monothiol glutaredoxins: Grx3, 4, and 5. Absence Grx5 leads to constitutive oxidative damage, exacerbating that caused by external oxidants. Phenotypic defects associated with the absence are suppressed overexpression ofSSQ1 ISA2, two genes involved in synthesis assembly iron/sulfur clusters into proteins. localizes at mitochondrial matrix, like other proteins these clusters, mature form lacks first 29 amino acids translation product. causes: 1) iron...
ABSTRACT Amphotericin B (AMB) is an antifungal drug that binds to ergosterol and forms pores at the cell membrane, causing loss of ions. In addition, AMB induces accumulation reactive oxygen species (ROS), although these molecules have multiple deleterious effects on fungal cells, their specific role in action mechanism remains unknown. this work, we studied ROS AMB. We determined intracellular induction 44 isolates different pathogenic yeast ( Candida albicans , parapsilosis glabrata...
We have analyzed the proteins that are oxidatively damaged when Saccharomyces cerevisiae cells exposed to stressing conditions. Carbonyl groups generated by hydrogen peroxide or menadione on of aerobically respiring were detected Western blotting, purified, and identified. Mitochondrial such as E2 subunits both pyruvate dehydrogenase α-ketoglutarate dehydrogenase, aconitase, heat-shock protein 60, cytosolic fatty acid synthase (α subunit) glyceraldehyde-3-phosphate major targets. In addition...
Glutaredoxins are members of a superfamily thiol disulfide oxidoreductases involved in maintaining the redox state target proteins. In Saccharomyces cerevisiae, two glutaredoxins (Grx1 and Grx2) containing cysteine pair at active site had been characterized as protecting yeast cells against oxidative damage. this work, another subfamily (Grx3, Grx4, Grx5) that differs from first single residue putative is described. This trait also characteristic for number bacteria to humans, with which...
Grx3 and Grx4, two monothiol glutaredoxins of Saccharomyces cerevisiae, regulate Aft1 nuclear localisation. We provide evidence a negative regulation activity by Grx4. The Grx domain both proteins played an important role in translocation to the cytoplasm. This function was not, however, dependent on availability iron. Here we demonstrate that Grx3, Grx4 interact each other vivo vitro, which suggests existence functional protein complex. Interestingly, interaction occurred independently...
A promoter-substitution cassette has been constructed that allows one-step substitution of chromosomal gene promoters for the tetracycline-regulatable tetO promoter in yeast cells, which uses kanMX4 as selective marker geneticin resistance. Oligonucleotides PCR amplification are designed to allow homologous recombination through short flanking regions homology with upstream sequences gene, upon transformation target cells. By testing three essential genes chromosome XV (YOL135c, YOL142w and...
The TOR (target of rapamycin) pathway controls cell growth in response to nutrient availability eukaryotic cells. Inactivation function by rapamycin or exhaustion is accompanied triggering various cellular mechanisms aimed at overcoming the stress. Here we report that inSaccharomyces cerevisiae protein kinase C (PKC)-mediated mitogen-activated regulated because upon specific Tor1 and Tor2 inhibition rapamycin, Mpk1 activated rapidly a process mediated Sit4 Tap42. Osmotic stabilization plasma...
In this study we analyze the participation of PKC1-MAPK cell integrity pathway in cellular responses to oxidative stress Saccharomyces cerevisiae. Evidence is presented demonstrating that only Pkc1 and upstream elements are essential for survival upon treatment with two oxidizing agents, diamide hydrogen peroxide. Mtl1 characterized first time as a cell-wall sensor stress. We also show actin cytoskeleton target Both peroxide provoke marked depolarization cytoskeleton, being Mtl1, Rom2...
In the present study, we have analyzed role of molecular chaperone Hsp60 in protection Saccharomyces cerevisiae against oxidative damage. We constructed mutant strains which levels protein, compared with wild-type cells, were four times greater, and addition doxycycline gradually reduces them to 20% wild-type. Under oxidative-stress conditions, progressive decrease these mutants resulted reduced cell viability an increase both peroxide species protein carbonyl content. Protection...
We have analyzed the heat stress response in yeast Saccharomyces cerevisiae by determining mRNA levels and transcription rates for whole transcriptome after a shift from 25°C to 37°C. Using an established mathematical algorithm, theoretical decay also been calculated experimental data. verified predictions selected genes their at different times during using regulatable tetO promoter. This study indicates that shock is not only due changes rates, but stabilities. stability affected 62% of it...
Glutaredoxins (GRXs) catalyze the reduction of protein disulfide bonds using glutathione as a reductant. Certain GRXs are able to transfer iron-sulfur clusters other proteins. To investigate function Arabidopsis (Arabidopsis thaliana) GRXS17, we applied strategy combining biochemical, genetic, and physiological approaches. GRXS17 was localized in nucleus cytosol, its expression elevated shoot meristems reproductive tissues. Recombinant bound Fe2S2 clusters, property likely contributing...
Grx5 is a yeast mitochondrial protein involved in iron-sulfur biogenesis that belongs to recently described family of monothiolic glutaredoxin-like proteins. No member this has been biochemically characterized previously. contains conserved cysteine residue (Cys-60) and non-conserved one (Cys-117). In work, we have purified wild type mutant C60S C117S proteins their biochemical properties. A redox potential –175 mV was calculated for Grx5. The pK values obtained by titration with...
Purified walls from Saccharomyces cerevisiae were treated chemically to release intrinsic mannoproteins. Boiling in 2% SDS gave the best results, although treatment 6 M-urea at room temperature also released significant amounts of mannoprotein radioactivity. Triton X-100, sodium deoxycholate and EDTA poor solubilizers. Electrophoretic patterns SDS- or urea-released mannoproteins SDS-acrylamide gels indicated a great heterogeneity molecular species, with more than 60 bands. Zymolyase,...
Glutaredoxins are thiol oxidoreductases that regulate protein redox state. In Saccharomyces cerevisiae, Grx1 and Grx2 cytosolic dithiol glutaredoxins, whereas Grx3, Grx4, Grx5 monothiol glutaredoxins. locates at the mitochondrial matrix is needed for iron/sulfur cluster biogenesis. Its absence causes phenotypes such as inactivation of enzymes sensitivity to oxidative stress. Whereas contains a single glutaredoxin domain, in Grx3 Grx4 thioredoxin-like domain fused domain. Here we have shown...
The Saccharomyces cerevisiae genome encodes three proteins that display similarities with human GSTOs (Omega class glutathione S-transferases) hGSTO1-1 and hGSTO2-2. yeast have been named Gto1, Gto2 Gto3, their purified recombinant forms are active as thiol transferases (glutaredoxins) against HED (β-hydroxyethyl disulphide), dehydroascorbate reductases dimethylarsinic acid reductases, while they not the standard GST substrate CDNB (1-chloro-2,4-dinitrobenzene). Their glutaredoxin activity...
The oxidative stress response in Saccharomyces cerevisiae has been analyzed by parallel determination of mRNA levels and transcription rates for the entire genome. A mathematical algorithm adapted a dynamic situation such as to stress, calculate theoretical decay from experimental data. Yeast genes have grouped into 25 clusters according level rate kinetics, average calculated each cluster. In most genes, changes one or both experimentally determined parameters occur during response. 24% are...