Project description:BY4741 (S288c haploid) cells have different gene expression in response to 0.4mM H2O2 when pretreated with 0.7M NaCl compared to cells that are not pretreated with NaCl (mock cells). Cells lacking Nup42p do not exhibit a different gene expression in response to H2O2 between naive cells and cells pretreated with 0.7M NaCl
Project description:To investigate the effects of DSF on Arabidopsis in Xcc infecction, we used 2 μM DSF pretreated the roos of Arabidopsis for 48h and then inoculated with Xcc, We then performed gene expression profiling analysis using data obtained from RNA-seq of pretreated/unpretreated at two time points (inoculated with Xcc or not)
Project description:BY4741 (S288c haploid) cells have different gene expression in response to 0.4mM H2O2 when pretreated with 0.7M NaCl compared to cells that are not pretreated with NaCl (mock cells). BY4741 cells were grown in log phase for 3 doublings (t0) and then the genomic expression of mock cells to 0.4mM H2O2 was assessed every 10 mins for 40 mins. To study the effects of NaCl treatement on H2O2 response, BY4741 cells were grown in log phase for 3 doublings (t0) and then the genomic expression of cells was measured in response to 0.7M NaCl over the course of 60 min every 15 mins. The cells were then removed from stress and grown in stress free media for four hours (T240). Then these cells were exposed to 0.4mM H2O2 and the genomic expression was measured every 10 minutes during the H2O2 timecourse of 40 minutes. To see if the handling of cells had an affect on gene expression, BY4741 cells were grown for exponentially for 3 doublings (t0), received a mock YPD treatment for 60 mins, grown for 4 hours (T240) and then collected to asses genomic expression. Duplicates were done for at 30 and 45 minutes for the NaCl timecourse, triplicate were done for 0, 10 and 20 minutes in H2O2 timecourse and duplicates were done for 30 and 40 minutes in the H2O2 timecourse.
Project description:Green plants are more robust to hydrogen peroxide (H2O2) stress and contain high endogeneous H2O2 levels which is generated during photorespiration and photosynthesis. Therefore, exgeneous H2O2 application mostly impose oxidative stress. To reduce endogenous H2O2 background, we adopted a strategy which is to grow Arabidopsis seedlings in the dark to eliminate light-induced H2O2 production, thus to reduce the endogenous H2O2 level. Exogenous H2O2 was then applied to induce transcriptome changes. Global gene expression is studied and compared between samples collected under 7d dark, 7d H2O2 treatment under dark and 7d light conditions.
2012-09-05 | GSE40574 | GEO
Project description:RNA-seq of the cold-pretreated Larimichthys crocea
Project description:In this project, we profiled the dynamics of proteome during Arabidopsis early embryogenesis using nanoproteomics. In addition, we identified some proteins which may be important during this process. Combining with RNA-seq, we unveiled the relationship between RNA and proteins during Arabidopsis early embryogenesis.
Project description:Green plants are more robust to hydrogen peroxide (H2O2) stress and contain high endogeneous H2O2 levels which is generated during photorespiration and photosynthesis. Therefore, exgeneous H2O2 application mostly impose oxidative stress. To reduce endogenous H2O2 background, we adopted a strategy which is to grow Arabidopsis seedlings in the dark to eliminate light-induced H2O2 production, thus to reduce the endogenous H2O2 level. Exogenous H2O2 was then applied to induce transcriptome changes. Global gene expression is studied and compared between samples collected under 7d dark, 7d H2O2 treatment under dark and 7d light conditions. We cultured seedlings in the dark to reduce endogenous H2O2. Three conditions were used for transcriptome profiling: dark grown (dark); dark grown with exogenous H2O2 treatment (H2O2); and light grown (light). Three types of conditions were used for Arabidopsis seedling culture: dark, dark with 5 mM H2O2 treatment and light. Each condition was performed with two biological replicates. The seedlings were harvested at 7 days old.
Project description:Hydrogen peroxide (H2O2) is an important messenger molecule for diverse cellular processes. H2O2 oxidizes proteinaceous cysteinyl thiols to sulfenic acid, also known as S-sulfenylation, thereby affecting the protein conformation and functionality. Although many proteins have been identified as S-sulfenylation targets in plants, site-specific mapping and quantification remain largely unexplored. By means of peptide-centric chemoproteomics, 1,537 S-sulfenylated sites were mapped on more than 1,000 proteins in Arabidopsis thaliana cells. The H2O2 sensitivity was quantified of more than 70% of these endogenous oxidation events toward exogenous H2O2 stimulation. Proteins involved in RNA and metabolic processing were identified as hotspots for S-sulfenylation. Moreover, S-sulfenylation frequently occurred on cysteines located in catalytic sites of enzymes or on cysteines involved in metal binding, hinting at direct mode-of-actions for redox regulation. Comparison of human and Arabidopsis S-sulfenylation datasets provided 155 conserved S-sulfenylated cysteines, including Cys181 of the Arabidopsis MITOGEN-ACTIVATED PROTEIN KINASE4 (AtMAPK4) that corresponds to Cys161 in the human MAPK1, which is speculated to be a redox-regulatory site. Replacement of the noncatalytic Cys181 of the recombinant AtMAPK4 by the redox-insensitive serine decreased the protein kinase activity, emphasizing the importance of this noncatalytic cysteine. Altogether, we quantitatively mapped the S-sulfenylated cysteines in Arabidopsis plants under oxidative stress and delivered an unprecedented inventory for unraveling the precise role of these oxidized cysteines in plant redox signaling.
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. To unravel the molecular mechanisms that regulate the response towards an impact of increased H2O2 levels in plant cells, we focused on the photorespiration-dependent peroxisomal H2O2 production in Arabidopsis thaliana mutants lacking CATALASE2 (CAT2) activity (cat2-2).
Project description:The plant hormone abscisic acid (ABA) modulates a number of plant developmental processes and responses to stress. In planta, ABA has been shown to induce reactive oxygen species (ROS) production through the action of plasma membrane-associated NADPH oxidases. Although quantitative proteomics studies have been performed to identify ABA- or hydrogen peroxide (H2O2)-dependent proteins, little is known about the ABA- and H2O2-dependent microsomal proteome changes. Here, we examined the effect of 50 µM of either H2O2 or ABA on the Arabidopsis microsomal proteome using tandem mass spectrometry and identified 86 specifically H2O2-dependent and 52 specifically ABA-dependent proteins that are differentially expressed. We observed differential accumulation of proteins involved in the TCA cycle notably in response to H2O2. Of these, aconitase 3 responded to both H2O2 and ABA. Additionally, over 30 proteins linked to RNA biology responded significantly to both treatments. Gene ontology categories such as ‘response to stress’ and ‘transport’ were enriched, suggesting that H2O2 or ABA directly and/or indirectly cause complex and partly overlapping cellular responses.