Project description:The transcriptomic changes induced in the human liver cell line HepG2 by 100µM menadione, 200µM TBH or 50µM H2O2 after treatment for 0.5, 1, 2, 4, 6, 8 and 24h.
Project description:We studied transcriptional responses of the fission yeast Schizosaccharomyces pombe to different intensities of H2O2 stress as well as two other oxidants: menadione sodium bisulfite and tert-butyl hydroperoxide. DNA microarrays were used to study the changes in expression profiles and the transcriptional regulation. We compare and contrast global expression and regulation of different intensities of H2O2 stress and different oxidants. The transcriptional response to low doses of H2O2 is very similar to menadione sodium bisulfite and the genes were controlled primarily by the transcription factor Pap1. The transcriptional response to medium and high doses of H2O2 is very similar to tert-butyl hydroperoxide and the genes were mostly regulated by the stress-activated MAP kinase Sty1p and the transcription factor Atf1p. We also compare global regulation of oxidative stress genes in fission and budding yeasts and discuss evolutionary implications.
Project description:The transcriptomic changes induced in the human liver cell line HepG2 by 100M-BM-5M menadione, 200M-BM-5M TBH or 50M-BM-5M H2O2 after treatment for 0.5, 1, 2, 4, 6, 8 and 24h. The study investigated differential gene expression in HepG2 cell line mRNA following 0.5, 1, 2, 4, 6, 8 and 24h hours of exposure to 100M-BM-5M menadione, 200M-BM-5M TBH or 50M-BM-5M H2O2 and medium without compound. Three biological replicates per compound/solvent. In total 126 arrays.
Project description:The early signaling events involved in oxidant recognition and triggering of oxidant-specific defense mechanisms to counteract oxidative stress still remains largely elusive. Our discovery driven comparative proteomics analysis revealed unique early signaling response of yeast Saccharomyces cerevisiae on proteome level to oxidants with different mechanism of action as early as 3 min after treatment with four oxidants, namely H2O2, cumene hydroperoxide (CHP), menadione and diamide, when protein abundances were compared using label-free quantification relying on a high-resolution mass analyzer (Orbitrap). We identified significant regulation of 196 proteins in response to H2O2, 569 proteins in response to CHP, 369 proteins in response to menadione and 207 proteins in response to diamide. Only 17 proteins were common across all treatments, but several more proteins were shared between two or three oxidants. Pathway analyses revealed that each oxidant triggered unique signaling mechanism associated with cell survival and repair. Signaling pathways mostly regulated by oxidants were Ran, mTOR, Rho, and eIEF2. Furthermore, each oxidant regulated these pathways in a unique way indicating specificity of response to oxidants having different mode of action. We hypothesize that interplay of these signaling pathway may be important in recognizing different oxidants to trigger different downstream MAPK signaling cascades and induce specific responses.
Project description:Background: During gut colonization, the enteric pathogen C. jejuni has to surmount the toxic effects of reactive oxygen species produced by its own metabolism, by the host immune system and by the intestinal microflora. Elucidation of C. jejuni oxidative stress defense mechanisms is critical for understanding Campylobacter pathophysiology. Results: The mechanisms of oxidative stress defenses in Campylobacter jejuni were characterized by transcriptional profiling, genes mutagenesis, and phenotypic analysis. The transcriptome changes, in response to H2O2, cumene hydroperoxide, or menadione exposure, were found to be oxidant specific and revealed the differential expression of genes belonging to a variety of biological pathways, from the classical oxidative stress defense systems, to the heat shock response, DNA repair and metabolism, fatty acid and capsule biosynthesis, and multidrug efflux pumps. To define the peroxide sensing regulator PerR, an isogenic mutant was constructed and its transcriptome profile compared to the wild-type strain. Sixty-six genes were found to belong to the PerR regulon. PerR appear to regulate gene expression both dependently and independently of the presence of iron and/or H2O2. The perR mutant was affected in its motility and attenuated in the chick colonization model. Mutagenic and phenotypic studies of the superoxide disumutase SodB, the alkyl-hydroxyperoxidase AhpC, and the catalase KatA, revealed their role in oxidative stress defenses and chick gut colonization. Conclusion: This study reveals the interplay between PerR, the iron metabolism and the oxidative stress defenses and highlights their role in the colonization and/or survival of C. jejuni in the chick cecum. Keywords: Transcriptional response to 3 oxidants (H2O2, menadione and cumene hydroperoxide) and characterization of the perR regulon (comparison of the transciptomes from the wild-type and perR mutant). To investigate the transcriptional responses of C. jejuni to oxidant exposure, hydrogen peroxide (H2O2), cumene hydroperoxide (CHP), or menadione sodium bisulfite (MND) was added to the 50 ml broth at a final concentration of 1 mM. The same amount of water or DMSO was added to the bacterial culture that served as reference samples for the transcriptional profile study in response to H2O2, MND or CHP. Furthermore, to investigate the transcriptional response of C. jejuni to H2O2 exposure in the presence of excess iron, ferrous sulfate was added to the bacterial culture at a final concentration of 40 µM, 15 min prior to H2O2 exposure. Ten minutes after the addition of the oxidant, total RNA was extracted and processed for microarray hybridization. To identify the PerR regulon, the wild-type strain C. jejuni NCTC 11168 and the perR mutant were grown in 500 ml flasks containing 250 ml of MEMα medium. At mid-log phase, 50 ml of the cultures were transferred to 100 ml flasks and ferrous sulfate and/or H2O2 were added . Ten minutes following the addition of H2O2 the cells were collected and the total RNA extracted.
Project description:Oxidative stress caused by Menadione or Hydrogen peroxide in synchronized Saccharomyces cerevisiae cultures. Alpha factor synchronized cultures (0.2-0.4 OD), treated at the beginning of S phase (25 min after release from G1 arrest) with either 2 mM Menadione (MD) or 0.24 mM Hydrogen peroxide (HP), show cell cycle effects. Cells treated with MD arrested at G1. Cells treated with HP delayed at S and then, after removal of HP at 135 minutes , continued the cell cycle, only to arrest at G2/M. Growth was carried out in 30C with shaking (295 rpm). Two time course experiments were performed with each oxidative stress agent, designated as H2O2 and H2O2_II, MD and MD_II. Keywords = oxidative stress Keywords = menadione Keywords = hydrogen peroxide Keywords = time course Keywords = cell cycle Keywords = yeast Keywords: other
Project description:Oxidative stress caused by Menadione or Hydrogen peroxide in synchronized Saccharomyces cerevisiae cultures. Alpha factor synchronized cultures (0.2-0.4 OD), treated at the beginning of S phase (25 min after release from G1 arrest) with either 2 mM Menadione (MD) or 0.24 mM Hydrogen peroxide (HP), show cell cycle effects. Cells treated with MD arrested at G1. Cells treated with HP delayed at S and then, after removal of HP at 135 minutes , continued the cell cycle, only to arrest at G2/M. Growth was carried out in 30C with shaking (295 rpm). Two time course experiments were performed with each oxidative stress agent, designated as H2O2 and H2O2_II, MD and MD_II. Keywords = oxidative stress Keywords = menadione Keywords = hydrogen peroxide Keywords = time course Keywords = cell cycle Keywords = yeast
Project description:We performed a broad-spectrum microarray analysis using mRNA from 3 NL and 5 DS astrocyte cultures. The microarray output was validated for several genes To explore the relation between gene expression and potential oxidative stress in DS samples, we also evaluated the transcription profile in NL astrocytes subjected to mild oxidative stress (50µM H2O2 for 24 hr). Samples of total mRNA from NL cultures non treated (NT), normal cultures treated with H2O2 (T), DS cultures NT and DS cultures T were included in the arrays.
Project description:We have developed QMP-SOs, a new class of superoxide-specific probes capable of proximity labelling and chemoproteomic profiling of proteins associated with superoxide redox biology. By coupling QMP-SOs with quantitative proteomics using tandem mass tag, we have established a chemoproteomics platform, QMP-SO-TMT. This enables us to profile proteins associated with superoxide biology in HepG2 cells treated with menadione, which induces mitochondrial superoxide production.
Project description:Expression Profiles of HepG2 cells treated with following compounds: Azathriopine, Furan, Tetradecanoyl phorbol acetate, Tetrachloroethylene, Diazinon and Dmannitol