Identification of ENO2-targeted downstream genes by ChIP-seq in Arabidopsis
Ontology highlight
ABSTRACT:
ORGANISM(S): Arabidopsis thaliana
PROVIDER: GSE151305 | GEO | 2022/06/04
REPOSITORIES: GEO
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Project description:Hydrogen cyanide (HCN) is co-produced with ethylene in plant cell and enzymatically detoxified mainly by the mitochondrial ß-cyanoalanine synthase (CAS-C1). Permanent or transient depletion of CAS-C1 activity in Arabidopsis results in physiological alteration in the plant that point to the function of HCN as a gasotransmitter molecule. Label-free quantitative proteomic analysis of enriched mitochondrial samples isolated from wild type and cas-c1 mutant reveled significant changes in protein content, identifying 451 proteins that are absent or less abundant in cas-c1 and 353 proteins only present or more abundant in the mutant background. Gene onthology classification of these proteins highlights proteomic changes that explains the root hairless phenotype and the altered inmune response observed in cas-c1 mutant. The mechanism of action of cyanide as signaling molecule has been addressed by two proteomic approaches focused on identifying S-cyanylation of cysteine as a posttranslational modification of proteins. Both the 2-imino-thiazolidine chemical method and direct untargeted analysis of proteins by LC-MS/MS identified a set of 163 proteins susceptibles to be S-cyanylated that include sedoheptulose 1, 7 biphosphatase (SBPase), the peptidyl-prolyl cis-trans isomerase (CYP20-3) and enolase 2 (ENO2). In vitro analysis of these proteins identified this modification in SBPase Cys74, CYP20-3 Cys259 and ENO2 Cys346 residues that affected the enzymatic activity of the enzymes. GO classification and protein-protein interaction cluster analysis revealed the function of S-cyanylation in the regulation of primary metabolic pathways as glycolysis, and the Calvin and S-adenosylmethionine cicles.
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Project description:In natural habitats, plants are often exposed to multiple stresses. Most studies, however, for plant abiotic stress responses analyzed those to individual stress but not combined stresses. In this report, we performed comparison analyses of gene expression to individual stresses, salt, osmotic and heat, and to a combination of these three stresses, which mimics arid conditions. We show here that the combined stress treatment induces unique gene expression pattern but not a simple reflection of the additive effects of individual stresses. First, the number of genes induced by combined stresses (150 mM NaCl, 200 mM mannitol and 35°C heat) was much smaller when compared to the sum of those induced by individual stress treatments, while the number of genes downregulated by multiple stresses was larger. A large number of genes induced by mannitol were not induced by multiple stresses, while those induced by salts were less affected in combined stress treatments. In addition, 125 genes, including13 for transcription factors, were found to be induced specifically by combined stress treatments. We report here that the plant response to a multi-stress environment represents an output of complex interactions between different stress aspects and signaling events of the various components of this environmental situation, and the determinative factor in this response is to avoid/minimize the antagonistic effects and to magnify the synergistic features of the imposed environmental challenges. Based on our results, we propose that genes that are highly induced by multiple treatments may be candidate for engineering stress tolerant crop plants. Wild-type plants of Arabidopsis thaliana were treated with three abiotic stresses, high salinity (150mM and 300mM), high osmotic pressure (200mM and 400mM mannitol) and heat (35°C), individually or simultaneously. Plants were treated with salt or mannitol for 16 hrs (first 1 hr and last 7 hrs are in light and other time in dark) or with heat for 4 hrs (in light) before sampling. There are three biologcal replicates.
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