Hydrogen Peroxide Regulation of Root Growth and Stem Cell Differentiation by XAANTAL1 in Arabidopsis thaliana
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ABSTRACT:
ORGANISM(S): Arabidopsis thaliana
PROVIDER: GSE262756 | GEO | 2024/07/20
REPOSITORIES: GEO
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Project description:Excessive levels of reactive oxygen species (ROS) cause cellular stress through damage to all classes of macromolecules and result in cell death. However, ROS can also act as signaling molecules in various biological processes. In plants, ROS signaling has been documented in environmental stress perception, plant development and cell death amongst others. Knowledge on the regulatory events governing ROS signal transduction is however still scratching the surface. To further elucidate the transcriptional response and regulation upon ROS accumulation we supplemented Arabidopsis seedlings with a 10mM hydrogen peroxide (H2O2) solution to trigger oxidative stress. After growth of 7 days, hydrogen peroxide (H2O2) was added to a final concentration of 10mM. Control plants were treated with the same volume of H2O. Seedlings were grown for 24h under the same controlled conditions. Design: 3 replicates x 2 conditions (7+1 day H2O or 7+1 day H2O2)
2016-06-10 | E-GEOD-80200 | biostudies-arrayexpress
Project description:Aim: To determine the effect of an AtrbohC mutation on the gene expression pattern in primary root tissue, to identify candidate genes acting downstream of AtrbohC, particularly any encoding antioxidant-related proteins, signal transduction components or proteins known to be required for normal root-hair development. Background: Root-hairs are a model system for investigating plant cell polarity. The root-hair mutant rhd2 (Schiefelbein and Somerville, 1990. Plant Cell, 2:235) has short hairs that burst at their tips, (Jones and Smirnoff, unpublished). RHD2 has been cloned and is identical to AtrbohC (L. Dolan, pers. comm.), which encodes a homologue of the superoxide-generating neutrophil respiratory burst oxidase catalytic subunit gp91phox (Torres et al., 1998. Plant J., 14:365). Superoxide rapidly dismutates to hydrogen peroxide (H2O2), suggesting that the rhd2 phenotype may result from reduced H2O2 levels in root-hair cells. Low doses of exogenous antioxidants phenocopy the rhd2 root-hair phenotype in wild-type plants (Jones and Smirnoff, unpublished) further supporting a role for H2O2 in root-hair growth. Fluorescent dyes that detect H2O2 show distinct localisation patterns in growing root-hair cells, (Jones and Smirnoff, unpublished). H2O2 may be an important second messenger in plant cell signalling with proposed roles in the development of cotton fibres (Potikha et al., 1999. Plant Physiol., 119: 849) and in ABA-induced stomatal closure (Zhang et al., 2001. Plant Physiol., 126: 1438). In cultured Arabidopsis cells H2O2 induces gene expression, including that of a gp91phox homologue, (Desikan et al., 1998. J. Exp. Bot., 49: 1767; Desikan, et al., 2000. Free Rad. Biol. Med., 28: 773; Baxter-Burrell et al., 2002. Science, 296: 2026) and activates a MAP kinase cascade (Desikan et al., 1999. J. Exp Bot., 50: 1863). cDNA microarray technology has been used previously to examine the effects of H2O2 on gene expression during oxidative stress (Desikan et al., 2001. Plant Physiol., 127: 159). We wish to investigate the effects of H2O2 on gene expression during root development using the rhd2 mutant. We are currently determining the expression pattern of RHD2. By extracting RNA from the small region of the primary root (for wild-type and rhd2 plants grown in sterile conditions) where root hairs are growing we hope to enrich for root-hair RNAs. This may reveal candidate genes that could be examined more closely at the single-cell level. This approach will provide new insights into the role of H2O2 in root-hair development.
2022-11-21 | E-NASC-54 | biostudies-arrayexpress
Project description:Aim: To determine the effect of an AtrbohC mutation on the gene expression pattern in primary root tissue, to identify candidate genes acting downstream of AtrbohC, particularly any encoding antioxidant-related proteins, signal transduction components or proteins known to be required for normal root-hair development. Background: Root-hairs are a model system for investigating plant cell polarity. The root-hair mutant rhd2 (Schiefelbein and Somerville, 1990. Plant Cell, 2:235) has short hairs that burst at their tips, (Jones and Smirnoff, unpublished). RHD2 has been cloned and is identical to AtrbohC (L. Dolan, pers. comm.), which encodes a homologue of the superoxide-generating neutrophil respiratory burst oxidase catalytic subunit gp91phox (Torres et al., 1998. Plant J., 14:365). Superoxide rapidly dismutates to hydrogen peroxide (H2O2), suggesting that the rhd2 phenotype may result from reduced H2O2 levels in root-hair cells. Low doses of exogenous antioxidants phenocopy the rhd2 root-hair phenotype in wild-type plants (Jones and Smirnoff, unpublished) further supporting a role for H2O2 in root-hair growth. Fluorescent dyes that detect H2O2 show distinct localisation patterns in growing root-hair cells, (Jones and Smirnoff, unpublished). H2O2 may be an important second messenger in plant cell signalling with proposed roles in the development of cotton fibres (Potikha et al., 1999. Plant Physiol., 119: 849) and in ABA-induced stomatal closure (Zhang et al., 2001. Plant Physiol., 126: 1438). In cultured Arabidopsis cells H2O2 induces gene expression, including that of a gp91phox homologue, (Desikan et al., 1998. J. Exp. Bot., 49: 1767; Desikan, et al., 2000. Free Rad. Biol. Med., 28: 773; Baxter-Burrell et al., 2002. Science, 296: 2026) and activates a MAP kinase cascade (Desikan et al., 1999. J. Exp Bot., 50: 1863). cDNA microarray technology has been used previously to examine the effects of H2O2 on gene expression during oxidative stress (Desikan et al., 2001. Plant Physiol., 127: 159). We wish to investigate the effects of H2O2 on gene expression during root development using the rhd2 mutant. We are currently determining the expression pattern of RHD2. By extracting RNA from the small region of the primary root (for wild-type and rhd2 plants grown in sterile conditions) where root hairs are growing we hope to enrich for root-hair RNAs. This may reveal candidate genes that could be examined more closely at the single-cell level. This approach will provide new insights into the role of H2O2 in root-hair development. Keywords: strain_or_line_design
2007-01-22 | GSE6165 | GEO
Project description:Thiol peroxidase (TpxD) is one of the key enzymes used to cope with oxidative stress conditions in S. pneumoniae. Previously, we demonstrated that TpxD expression is modulated in response to H2O2 and is involved in the regulation of the psa operon. In the current study, we focus on TpxD function as a sensor and global regulator in the bacterial response to H2O2. By using microarray assays, we show that TpxD expression is necessary for the activation of bacterial global gene response to H2O2. The mechanism underlying TpxD regulatory function was further elucidated by replacing the catalytic cysteine (Cys58) with alanine (Ala58) in TpxD. This point mutation prevented tpxD up-regulation by H2O2, as well as the effect of H2O2 on additional genes that were affected by H2O2 in the WT but not in ∆tpxD, signifying that this cysteine is crucial for TpxD signaling activity. We then tried to identify a candidate transcription factor which regulates tpxD expression under H2O2 stress. Bioinformatics analysis identified a putative CodY 15 bp binding site in the proximal upstream region of tpxD coding sequence. Genetic engineering techniques and EMSA assays confirmed the presence of an active CodY box. Indeed, no up-regulation of TpxD was noticed in ∆codY challenged with H2O2. These data identify CodY as the transcription factor modulating tpxD expression under H2O2 stress. We propose a model of gene regulation under oxidative stress conditions which places TpxD as an intermediary between H2O2 and the global bacterial response to oxidative stress. This SuperSeries is composed of the SubSeries listed below.
2017-07-27 | GSE65157 | GEO
Project description:Aim: To determine the effect of an AtrbohC mutation on the gene expression pattern in primary root tissue, to identify candidate genes acting downstream of AtrbohC, particularly any encoding antioxidant-related proteins, signal transduction components or proteins known to be required for normal root-hair development. Background: Root-hairs are a model system for investigating plant cell polarity. The root-hair mutant rhd2 (Schiefelbein and Somerville, 1990. Plant Cell, 2:235) has short hairs that burst at their tips, (Jones and Smirnoff, unpublished). RHD2 has been cloned and is identical to AtrbohC (L. Dolan, pers. comm.), which encodes a homologue of the superoxide-generating neutrophil respiratory burst oxidase catalytic subunit gp91phox (Torres et al., 1998. Plant J., 14:365). Superoxide rapidly dismutates to hydrogen peroxide (H2O2), suggesting that the rhd2 phenotype may result from reduced H2O2 levels in root-hair cells. Low doses of exogenous antioxidants phenocopy the rhd2 root-hair phenotype in wild-type plants (Jones and Smirnoff, unpublished) further supporting a role for H2O2 in root-hair growth. Fluorescent dyes that detect H2O2 show distinct localisation patterns in growing root-hair cells, (Jones and Smirnoff, unpublished). H2O2 may be an important second messenger in plant cell signalling with proposed roles in the development of cotton fibres (Potikha et al., 1999. Plant Physiol., 119: 849) and in ABA-induced stomatal closure (Zhang et al., 2001. Plant Physiol., 126: 1438). In cultured Arabidopsis cells H2O2 induces gene expression, including that of a gp91phox homologue, (Desikan et al., 1998. J. Exp. Bot., 49: 1767; Desikan, et al., 2000. Free Rad. Biol. Med., 28: 773; Baxter-Burrell et al., 2002. Science, 296: 2026) and activates a MAP kinase cascade (Desikan et al., 1999. J. Exp Bot., 50: 1863). cDNA microarray technology has been used previously to examine the effects of H2O2 on gene expression during oxidative stress (Desikan et al., 2001. Plant Physiol., 127: 159). We wish to investigate the effects of H2O2 on gene expression during root development using the rhd2 mutant. We are currently determining the expression pattern of RHD2. By extracting RNA from the small region of the primary root (for wild-type and rhd2 plants grown in sterile conditions) where root hairs are growing we hope to enrich for root-hair RNAs. This may reveal candidate genes that could be examined more closely at the single-cell level. This approach will provide new insights into the role of H2O2 in root-hair development. Experiment Overall Design: Number of plants pooled:100
2008-06-14 | E-GEOD-6165 | biostudies-arrayexpress
Project description:Hemoglobin (Hb) is released from red blood cells (RBC) during intravascular hemolysis. Cell free Hb has been implicated to play a pathogenic role in hemolytic diseases such as sickle cell anemia or malaria. Hydrogen peroxide (H2O2) is the most prevalent reactive oxygen species which is produced in excess during inflammation or tissue injury and has been included as a potential mediator of oxidative stress related cell and tissue damage. The biologic significance of Hbs peroxidase activity as an anti-oxidant is controversial as the radicals and higher oxidation iron species which are potentially released during these reactions could be a source of exaggerated oxidative damage. Using a cell culture model of low-flux continuous H2O2 generation by glucose-oxidase (GOx) Hb revealed the potential to protect vascular smooth muscle cells and macrophages from H2O2 mediated glutathione depletion and cell death. As revealed by gene array analysis, Hb effectively abolished H2O2 induced oxidative stress response in these cells. Through electron microscopy and quantitative mass spectrometry extensive oxidation of specific Hb amino acid residues, polymerization and precipitation was defined. This may be a consequence of potential protective mechanism within Hb. The net biologic effect of Hb, when present in oxidative rich environments, is likely a result of the balance of H2O2 consumption (protection) on one hand and the production of free radicals (damage) on the other hand. Further the extensive structural changes occurring during the reaction of ferrous Hb with H2O2 could 'absorb' a significant amount of oxidative impact and thereby further protect the environment from heme derived free radical damage. The intrinsic peroxidase activity of hemoglobin seems to be a constitutive 'enzymatic' function of Hb which adds a novel facet to the multivalent role played by the abundant oxygen carrier protein.
2009-09-15 | E-MEXP-2263 | biostudies-arrayexpress
Project description:PtomitAPX, which is a newly discovered ascorbate peroxidase (APX) specifically localized in the mitochondria of Populus tomentosa. Our antisense and overexpression studies are based on PtomitAPX. [Abstract] Plant mitochondria are major organelles of reactive oxygen species (ROS) production, are also targeted by ROS, which reduces mitochondrial efficiency and increases ROS production in a self-destructive cycle. Ascorbate peroxidase have pivotal roles in ROS-scavenging because even very low concentrations are sufficient for H2O2 decomposition We identified two ascorbate peroxidases, PtomitAPX and PtosAPX, are targeted to mitochondria in Populus tomentosa Carr. PtosAPX which is the homologous gene of arabidopsis thaliana sAPX, is also dual targeted to both chloroplast and mitochondria but is closer to chloroplastic isoform by ELISA and qPCR analysis. And PtomitAPX which is specifically targeted to mitochondria, is primary APX in mitochondria. The expression of PtomitAPX was significantly different from that of PtosAPX. PtomitAPX plays an important role in controlling the ROS balance and maintaining mitochondrial structure and function, thus regulating cell PCD and plant development in P opulus tomentosa Carr. The structure and function of mitochondria, particularly the efficiency of oxidative phosphorylation (OXPHOS), were impaired, cell PCD were increased and plant growth was retarded and partially restored by exogenous ASA in antisense PtomitAPX transgenic Populus,. The PtomitAPX expression level showed a strong relationship with ROS balance, mitochondrial structure, and plant growth. Our findings provide valuable insight into the unexplored mechanism of independent ROS balance by the ascorbate-glutathione cycle in mitochondria. Moreover, the data enhance understanding of the central role of mitochondria during plant growth, particularly in non-photosynthetic tissues of woody trees. Methods: Total RNA was isolated from wild type (WT), antisense (anti-3) and overexpression (OX) cells using TRIzol reagent according to the manufacturer’s protocol (Invitrogen). The samples were sequenced using an Illumina Genome Analyzer (HiSeq™ 2000; Illumina, San Diego, CA). The raw reads were data-filtered to obtain high-quality clean reads. The clean reads were mapped to the P. trichocarpa reference genome and reference genes using SOAPaligner/SOAP2. No more than two mismatches were allowed in the alignment. Gene expression levels were calculated as reads per kilobase per million reads (RPKM). Differential expression analysis among the WT, anti-3 and OX was performed using the DEGseq R package based on normalized read counts. A corrected P value of < 0.001 and |log2Ratio| > 1.5 were set as the threshold for significantly differential expression. Results: Compared with the WT, 3571 genes with significantly different expression levels (fold change > 1.5 or < −1.5, and corrected P < 0.001) were detected in anti-3 and 95.58% (3413) of them reverted or partial reverted to WT in anti-3 treated with 1 mM AsA. RNA-seq analysis of WT, overexpression, and overexpression cells treated with 10 mM H2O2, were also employed. Compared with the WT, 5542 genes with significantly different expression levels(fold change > 1.5 or < −1.5, and corrected P < 0.001) were detected in OX and 66.47% (3684) of them were reverted or partial reverted to WT in OX treated with 10 mM H2O2.
2018-10-23 | GSE121562 | GEO
Project description:Elevated reactive oxidative species (ROS) may be crucial for cellular transformation and tumor progression. Defining the response of stem cells to ROS is critical to understanding the unique sensitivity of stem cells to ROS-mediated transformation and tumor initiation. We examined both the immediate transcriptional and phospho-proteomic responses of murine ES cells induced by a single brief dose of hydrogen peroxide (H2O2) to create a more complete understanding of cell response to ROS. Experiment Overall Design: Murine ES cells were briefly exposed to low and high dose of H2O2 then allowed to recover for 1 hour in maintenance medium before RNA extraction and MG_U74Av2 microarray hybridization.
2009-11-10 | E-GEOD-18708 | 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:Myosins are evolutionarily conserved motor proteins that interact with actin filaments to regulate organelle transport, cytoplasmic streaming and cell growth. Plant specific Class XI myosin proteins direct cell division and root organogenesis. However, the roles of Class VIII myosin proteins in plant growth and development are less understood. Here, we investigated the function of an auxin-regulated Class VIII myosin, Arabidopsis thaliana Myosin 1 (ATM1), using genetics and live cell microscopy. ATM1 is a plasmodesmata-localized protein that is enriched in actively dividing cells in the root apical meristem (RAM). Loss of ATM1 function results in impaired primary root growth due to decreased RAM size and reduced cell proliferation in a sugar dependent manner. Examination of stem cell marker line expression in atm1-1 indicated impaired division of the lateral root cap and columella cells and a diminished auxin response. Transcriptome analysis of atm1-1 linked these growth defects to dysregulation of cell cycle and auxin pathway genes. Complementation of ATM1 loss-of- function mutant restored root growth and cell cycle progression in the root meristem. Collectively, these results provide novel evidence that ATM1 functions to influence cell proliferation and differentiation in primary roots in response to auxin and sugar cues.
2023-06-08 | GSE200917 | GEO