Effect of MnSOD knockout cells on gene profile expression
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE14855 | GEO | 2009/02/18
SECONDARY ACCESSION(S): PRJNA112137
REPOSITORIES: GEO
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Project description:MnSOD is an essential primary antioxidant enzyme that converts superoxide radicals and protons to hydrogen peroxide (H2O2) within the mitochondrial matrix, generated by respiratory chain activity We used microarrays of cells knocked down for MnSOD and a mock transfected cells as their control (siScramble) to reveal changes in gene expression profile WT-AML-12 cells silenced (si) for MnSOD were prepared, briefly: The backbone vector pSUPER.retro.puro was used for silencing. Transcripts of 60-nt long oligos constituting hairpin RNAs were designed to generate a low level of MnSOD silencing: a nine-loop (9 L) hairpin with 20 nt of MnSOD sense (siMnSOD) and a 9 L hairpin with 20 nt of scramble-no gene recognition control sense (Scr). Retroviral particles were produced by triple transfection of HEK 293T cells (ATCC No. CRL-11268T) with the retroviral vector (2 ug), pMD-gag-pol (1 ug) and pVSV-G (1 ug) using Trans-IT (Mirusbio Corporation, Madison, WI). For stable transfection, the various retroviruses were introduced with WT-AML-12 hepatocytes using polybrene (Sigma-Aldrich, St. Louis, MO, USA) in a six-well plate followed by selection with 10 ug puromycin for 2 weeks. To identify sets of genes that are differentially expressed in siMnSOD AML-12 cells compared to Scr-no sense control AML-12 cells, a Gene Chip approach was utilized. Total RNA was extracted (using TRI reagent) and purified using the RNeasy kit (Qiagen, Valencia, CA). Three different batches of frozen cells were taken to microarray analysis. Microarray analysis was performed on an Affymetrix Mouse Gene 1.0 ST chip containing 28,853 genes. The chips were processed and scaled at the Weizmann Institute of Science (Rehovot, Israel) using Affymetrix MAS5.
2009-02-18 | E-GEOD-14855 | biostudies-arrayexpress
Project description:In muscle, reactive oxygen species (ROS) generation increases with strenuous activity, chronic unloading, and inflammatory stimuli; skeletal muscle function is very sensitive to ROS; and there are redox-sensitive signaling pathways. Using myogenic cell cultures, we asked whether hydrogen peroxide (H2O2) induces adaptive changes in skeletal muscle gene expression. H2O2 downregulated or failed to induce antioxidant or apoptotic genes in the myotubes. Instead, H2O2 changed the expression of genes for cytosolic and mitochondrial enzymes, and upregulated inflammatory mediators. Finally, H2O2 had a mostly inhibitory effect on the expression of many transcription factors. The results indicate that mild oxidative stress may induce an adaptive response in skeletal muscle without antioxidant upregulation or apoptosis. Keywords: Gene Expression, C2C12 Myotubes, Oxidative Stress, Adaptation
2005-08-11 | GSE3078 | GEO
Project description:We studied the transcriptomic response of S. pneumoniae to moxifloxacin, a fluoroquinolone that inhibits DNA gyrase. At 10 × MIC, 30 and 140 responsive genes were detected at 15 and 30 minutes, respectively. Several pathways leading to an increase in pyruvate showed up-regulation. These included 2 genes introducing P-sugars into the glycolysis and 3 out of 7 genes of the glycolysis pathway, which converts fructose-6P to pyruvate. In addition, an increase in acetyl-coA would be expected from the down-regulation of the genes coding for the acetyl-coA carboxylase, and in turn, to a further increase in pyruvate by the up-regulation of formate acetyltransferase. Since pyruvate is converted to hydrogen peroxide (H2O2) by pyruvate oxidase (SpxB), its increase would lead to an equivalent increase in the intracellular amount of H2O2, and in turn, in those of hydroxyl radicals resulting from the Fenton reaction, which damage DNA, lipids and proteins. We observed similar increases in the production of H2O2 and hydroxyl radicals under moxifloxacin treatment. These reactive oxygen species contributed to the lethality of the drug, as showed by the attenuation of its lethality in a strain lacking SpxB. These results support the production of redox alterations by fluoroquinolones and are in agreement with our previous findings showing that levofloxacin, an inhibitor of topoisomerase IV, triggers the transcriptional activation of iron transport genes. Both fluoroquinolones stimulate the Fenton reaction in their mechanism of action, by increasing the amounts of any of their two components: iron by levofloxacin or H2O2 by moxifloxacin.
2016-02-05 | GSE68947 | GEO
Project description:Superoxide radical anion and other Reactive Oxygen Species are constantly produced during respiration. In mitochondria, the dismutation of the superoxide radical anion is accelerated by the mitochondrial superoxide dismutase 2 (SOD2), an enzyme that has been traditionally associated with antioxidant protection. However, increases in SOD2 expression promote oxidative stress, indicating that there may be a prooxidant role for SOD2. We show that SOD2, which normally binds manganese, can incorporate iron and generate an alternative isoform with peroxidase activity. The switch from manganese to iron allows FeSOD2 to utilize H2O2 to promote oxidative stress. We found that FeSOD2 is formed in cultured cells. FeSOD2 causes mitochondrial dysfunction and higher levels of oxidative stress in cultured cells. We show that formation of FeSOD2 converts an antioxidant defense into a prooxidant peroxidase that leads to cellular changes seen in multiple human diseases.
2017-12-05 | GSE106206 | GEO
Project description:Adaptation to hydrogen peroxide in Saccharomyces cerevisiae is profiled with expression arrays. Adaptation describes the process in which a mild dose of toxin (in this case, hydrogen peroxide) is able to protect against a later acute dose. Here, we study two adaptive protocols (0.1 mM H2O2 and 0.1 + 0.4 mM H2O2) and one acute protocol (0.4 mM H2O2) to identify processes uniquely involved in adaptation. Predictions from these studies are validated in expression profiling of deletion mutants of the transcription factors Yap1, Mga2, and Rox1.
2009-04-23 | GSE12602 | GEO
Project description:Fluorescent proteins are an important tool that has become omnipresent in life sciences research. They are frequently used for localization of proteins and monitoring of cells. Green fluorescent protein (GFP) was the first and has been the most used fluorescent protein. Enhanced GFP (eGFP) was optimized from wild-type GFP for increased fluorescence yield and improved expression in mammalian systems. Fluorescent proteins are expressed colorless and immature and, for eGFP, the conversion to the fluorescent form, mature, is known to produce one equivalent of hydrogen peroxide (H2O2) per molecule of chromophore. Even though it has been proposed that this process is non-catalytic and generates nontoxic levels of H2O2, we sought to investigate the role of fluorescent proteins in generating free radicals and inducing oxidative stress in biological systems.
2017-03-16 | GSE96671 | GEO
Project description:Oxidative Stress Protection and the Repair Response To Hydrogen Peroxide in the Hyperthermophilic Archaeon Pyrococcus furiosus Pyrococcus furiosus is a shallow marine, anaerobic archaeon that grows optimally at 100°C. Addition of H2O2 (0.5 mM) to a growing culture resulted in cessation of growth with a 2 hour lag before normal growth resumed. Whole genome transcriptional profiling revealed that the main response occurs within 30 min of peroxide addition, with the up-regulation of 62 open reading frames (ORFs), 36 of which are part of 10 potential operons. More than half of the up-regulated ORFs are of unknown function while some others encode proteins that are involved potentially in sequestering iron and sulfide, in DNA repair and in generating NADPH. This response is thought to involve primarily damage repair rather than protection, since cultures exposed to sub-toxic levels of H2O2 were not more resistant to the subsequent addition of H2O2 (0.5 – 5.0 mM). Consequently, there is little if any induced protective response to peroxide, rather, the organism maintains a constitutive protective mechanism involving high levels of oxidoreductase-type enzymes such as superoxide reductase, rubrerythrin and alkyl hydroperoxide reductase I. The related hyperthermophiles P. woesei and Thermococcus kodakaraensis were more sensitive to peroxide than P. furiosus, apparently due to the lack of several of its peroxide-responsive ORFs.
2010-02-25 | GSE20470 | GEO
Project description:Investigating the oxidative stress response: Candida glabrata strains were stressed with hydrogen peroxide and menadione (causing oxygen radicals) to induce the oxidative stress regulon, which is thought to be upregulated during the oxidative burst inside of phagocytic cells.
2011-01-01 | E-MEXP-2915 | biostudies-arrayexpress
Project description:Hydrogen peroxide (H2O2) can act as a signaling molecule that influences various aspects of plant growth and development, including stress signaling and cell death. Catalase deficient plants are pioneering systems which accumulate hydrogen peroxide (H2O2) from peroxisomal origin during photorespiratory challenges. Respiratory burst oxidase homologues D and F are known to participate in intracellular oxidative stress response launched in cat2 mutants (Chaouch et al., 2012). We studied the compared the transcriptional response of cat2 rbohD and cat2 rbohF double mutants versus the cat2 background to further adress their role during photorespiratory stress.
2016-06-10 | GSE81516 | GEO
Project description:To identify genes that regulate root development in a hydrogen peroxide devendent manner, we performed a time course microarray analysis of root treated with 1mM H2O2.
2018-07-31 | GSE85015 | GEO