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Arabidopsis AtbHLH112 protein positively regulates tolerance to abiotic stress and binds to the E-box and a novel motif GCG-box

ABSTRACT: Plant basic helix-loop-helix (bHLH) transcription factors are involved in physiological and developmental processes, and also play essential roles in abiotic stresses. However, their exact roles in abiotic stress are still need to be elucidated, and most of bHLHs have not been functionally characterized. In the present study, we characterized the functional role of AtbHLH112 in response to abiotic stresses. AtbHLH112 is a nuclear-localized protein, and its nuclear-localization is induced by salt, drought and ABA. Besides binding to E-box motif, AtbHLH112 is found to bind to a novel motif with the sequence “GG[GT]CC[GT][GA][TA]C” (GCG-box), and the binding affinity is induced by salt and ABA. Gain- and loss-of-function analyses showed that the transcript level of AtbHLH112 is positively correlated with salt and drought tolerance. AtbHLH112 mediates stress tolerance by upregulating the expression of P5CS genes and decreasing the expression of P5CDH and PRODH genes to increase proline levels, and via enhancing the expression of POD and SOD genes to improve ROS scavenging ability. All data together suggested that AtbHLH112 regulates the expression of genes through binding to GCG-box and E-box to mediate the physiological stress responses, including proline biosynthesis and ROS scavenging pathways to enhance stress tolerance.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE60260 | GEO | 2014/08/09

SECONDARY ACCESSION(S): PRJNA257827

REPOSITORIES: GEO

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Arabidopsis AtbHLH112 protein positively regulates tolerance to abiotic stress and binds to the E-box and a novel motif GCG-box

Project description:Plant basic helix-loop-helix (bHLH) transcription factors are involved in physiological and developmental processes, and also play essential roles in abiotic stresses. However, their exact roles in abiotic stress are still need to be elucidated, and most of bHLHs have not been functionally characterized. In the present study, we characterized the functional role of AtbHLH112 in response to abiotic stresses. AtbHLH112 is a nuclear-localized protein, and its nuclear-localization is induced by salt, drought and ABA. Besides binding to E-box motif, AtbHLH112 is found to bind to a novel motif with the sequence M-bM-^@M-^\GG[GT]CC[GT][GA][TA]CM-bM-^@M-^] (GCG-box), and the binding affinity is induced by salt and ABA. Gain- and loss-of-function analyses showed that the transcript level of AtbHLH112 is positively correlated with salt and drought tolerance. AtbHLH112 mediates stress tolerance by upregulating the expression of P5CS genes and decreasing the expression of P5CDH and PRODH genes to increase proline levels, and via enhancing the expression of POD and SOD genes to improve ROS scavenging ability. All data together suggested that AtbHLH112 regulates the expression of genes through binding to GCG-box and E-box to mediate the physiological stress responses, including proline biosynthesis and ROS scavenging pathways to enhance stress tolerance. Differentially expression genes of AtbHLH112-overexpression plants, mutant (SALK_033618C) plants and wild type of Columbia Arabidopsis thaliana were measured under salt stressed and normal condition for 3 hours, respectively. Three independent experiments were performed at each treatment using different plants for each experiment.

2014-08-09 | E-GEOD-60260 | biostudies-arrayexpress

Arabidopsis AtbHLH112 protein binds to the G-box and a novel motif GCG-box to regulate gene expression in response to abiotic stress

Project description:Plant basic helix-loop-helix (bHLH) proteins play essential roles in physiological and developmental processes and are also involved in abiotic stresses. However, their exact roles in abiotic stress are still not fully understood, and most of them have not been functionally characterised. In the present study, we characterised the functional role of AtbHLH112 in response to abiotic stress. A WRKY gene, AtWRKY66, can regulate the expression of the AtbHLH112 via binding to W-box motifs present in its promoter. AtbHLH112 is a nuclear-localised protein, and its nuclear localisation is increased upon exposure to NaCl, mannitol and ABA. In addition to binding to the G-box motif, AtbHLH112 is found to bind to a novel motif M-bM-^@M-^\GGGCCGGTCM-bM-^@M-^] (named the GCG-box) to regulate gene expression. Gain- and loss-of-function analyses showed that the transcript level of AtbHLH112 is positively correlated with tolerance to salt and drought. AtbHLH112 can confer stress tolerance via enhanced expression of POD and SOD genes to improve ROS scavenging ability and via upregulated expression of P5CS genes and decreased expression of P5CDH and PRODH genes to improve proline levels. Our data suggested that AtbHLH112 regulates the expression of genes via binding to the G-box and the GCG-box to improve stress-related pathways, such as ROS scavenging and proline biosynthesis. Differentially expression genes of AtbHLH112-overexpression plants and mutant (SALK_033618C) plants of Arabidopsis thaliana were measured under salt stressed and normal condition for 3 hours, respectively. Three independent experiments were performed at each treatment using different plants for each experiment.

2013-11-14 | E-GEOD-52326 | biostudies-arrayexpress

Arabidopsis AtbHLH112 protein binds to the G-box and a novel motif GCG-box to regulate gene expression in response to abiotic stress

Project description:Plant basic helix-loop-helix (bHLH) proteins play essential roles in physiological and developmental processes and are also involved in abiotic stresses. However, their exact roles in abiotic stress are still not fully understood, and most of them have not been functionally characterised. In the present study, we characterised the functional role of AtbHLH112 in response to abiotic stress. A WRKY gene, AtWRKY66, can regulate the expression of the AtbHLH112 via binding to W-box motifs present in its promoter. AtbHLH112 is a nuclear-localised protein, and its nuclear localisation is increased upon exposure to NaCl, mannitol and ABA. In addition to binding to the G-box motif, AtbHLH112 is found to bind to a novel motif “GGGCCGGTC” (named the GCG-box) to regulate gene expression. Gain- and loss-of-function analyses showed that the transcript level of AtbHLH112 is positively correlated with tolerance to salt and drought. AtbHLH112 can confer stress tolerance via enhanced expression of POD and SOD genes to improve ROS scavenging ability and via upregulated expression of P5CS genes and decreased expression of P5CDH and PRODH genes to improve proline levels. Our data suggested that AtbHLH112 regulates the expression of genes via binding to the G-box and the GCG-box to improve stress-related pathways, such as ROS scavenging and proline biosynthesis.

2013-11-14 | GSE52326 | GEO

ChIP-Sequencing of AL5 over-expressing Arabidopsis and its knock out mutant

Project description:Plant homeodomain (PHD) finger proteins affect growth and development by regulating transcription and reading epigenetic modifications of histones, but their functions in abiotic stress responses remain largely unclear. Here we characterize seven Arabidopsis thaliana Alfin1-like PHD finger proteins (ALs) in the response to abiotic stresses. ALs localize to the nucleus and repress transcription. Except AL6, all the ALs bind to G-box element. Changes of the amino acids at positions 34 and 35 in AL6 cause the loss of G-box binding ability. Expression of the ALs responded differently to osmotic stress, salt, cold and abscisic acid treatments. AL5 was induced by multiple stresses, and AL5-overexpressing plants showed higher tolerance to salt, drought and freezing stress than Col-0. Also, al5 mutants showed reduced stress tolerance. ChIP-Seq assay helps find the direct targets of AL5.

2014-04-12 | GSE56706 | GEO

ChIP-Sequencing of AL5 over-expressing Arabidopsis and its knock out mutant

2014-04-12 | E-GEOD-56706 | biostudies-arrayexpress

The C-terminal domains SnRK2-box and ABA-box have a role in sugarcane SnRK2s auto-activation and activity

Project description:Resistance to drought stress is fundamental to plant survival and development. Abscisic acid (ABA) is one of the major hormones involved in different types of abiotic and biotic stress responses. ABA intracellular signaling has been extensively explored in Arabidopsis thaliana and occurs via a phosphorylation cascade mediated by three related protein kinases, denominated SnRK2s (SNF1-related protein kinases). However, the role of ABA signaling and the biochemistry of SnRK2 in crop plants remains underexplored. Considering the importance of the ABA hormone in abiotic stress tolerance, here we investigated the regulatory mechanism of sugarcane SnRK2s - known as SAPKs (Stress/ABA-activated Protein Kinases). The crystal structure of ScSAPK10 revealed the characteristic SnRK2 family architecture, in which the regulatory SnRK2-box interacts with the kinase domain αC helix. To study sugarcane SnRK2 regulation, we produced a series of mutants for the protein regulatory domains SnRK2-box and ABA-box. Surprisingly, mutations in the SnRK2-box did not drastically affect sugarcane SnRK2 activity, in contrast to previous observations for the homologous proteins in Arabidopsis. Also, we found that the ABA-box might have a role in SnRK2 activation in the absence of PP2C phosphatase. Taken together, our results demonstrate that both C-terminal domains of sugarcane SnRK2 proteins play a role in protein activation and activity.

2019-11-13 | PXD014298 | Pride

Arabidopsis thaliana

Project description:Arabidopsis AtbHLH112 protein positively regulates tolerance to abiotic stress and binds to the E-box and a novel motif GCG-box

| PRJNA257827 | ENA

Transcriptomic and alternative splicing analyses reveal mechanisms of the difference in salt tolerance between barley and rice

Project description:Both barley (Hordeum vulgare) and rice (Oryza sativa) belong to Poaceae family, but differ greatly in salt tolerance. In order to understand molecular mechanisms in the difference of salt tolerance between the two species, the responses of transcriptomic profiles to salt stress were compared between rice (cultivar Nipponbare) and barley (accession XZ26) to reveal how alternative splicing (AS) coordinates with transcriptional regulation in adaptation to salt stress. Physiological study showed that XZ26 had higher salt tolerance than Nipponbare, as reflected by less growth inhibition, lower shoot Na+ concentration and higher K+/Na+ ratio when exposed to salt stress. Transcriptomic analysis showed that XZ26 had higher ROS scavenging ability, less degradation of protein kinases and enhanced anti-oxidation. Moreover, AS genes related to ion transporter genes and transcription factors could enhance and amplify K+/Na+ homeostasis and signal transduction cascades. We proposed that higher salt tolerance of barley accession XZ26 is attributed to its superior K+/Na+ homeostasis, tissue detoxication and less energy consumption. The present results provide insights at transcriptomic levels into reasons why barley has higher salt tolerance than rice.

2019-06-05 | GSE132183 | GEO

Comprehensive dissection of primary metabolites in response to diverse abiotic stress in barley at seedling stage

Project description:Plants will meet various abiotic stresses during their growth and development. One of the important strategies for plants to deal with the stress is involved in metabolic regulation, causing the dramatic changes of metabolite profiles. Metabolomic studies have been intensively conducted to reveal the responses of plants to abiotic stress, but most of them were limited to one or at most two abiotic stresses in a single experiment. In this study, we compared the metabolite profiles of barley seedlings exposed to seven abiotic stresses simultaneously, including drought, salt stress, aluminum (Al), cadmium (Cd), deficiency of nitrogen (N), phosphorus (P) and potassium (K). The results showed that metabolite profiles of barley under these stresses could be classified into three types: osmotic stresses (drought and salt); metal stresses (Al and Cd) and nutrient deficiencies (N, P and K deficiencies). Compared with the control, some metabolites (including polyamines, raffinose and piperonic acid) in plants exposed to all abiotic stresses changed significantly, while some other metabolites showed the specific change only under a certain abiotic stress, such as proline being largely increased by osmotic stress (drought and salinity), the P-containing metabolites being largely decreased under P deficiency, some amino acids (lysine, tyrosine, threonine, ornithine， glutamine and so on) showing the dramatic reduction in the plants exposed to N deficiencies, respectively. The current meta-analysis obtained a comprehensive view on the metabolic responses to various abiotic stress, and improved the understanding of the mechanisms for tolerance of barley to abiotic stress.

2021-01-07 | ST001648 | MetabolomicsWorkbench

Maize root LC-MSMS, part 2 - Comparative proteomics of contrasting maize genotypes provides insights into salt-stress tolerance mechanisms

Project description:Salt stress is one major abiotic stress limiting maize grain yield throughout the world.To better understand maize salt tolerance molecular mechanism, comparative proteomic analysis was conducted on seedling roots of salt-tolerant genotype Jing724 and salt-sensitive genotype D9H under NaCl. Jing724 exhibited significantly higher germination rate and growth parameters (weight/length) than did D9H under salt treatment.We identified 565 differentially regulated proteins (DRP) using iTRAQ and 89 were specific to Jing724 while 424 were specific to D9H. In salt stressed Jing724, pentose phosphate pathway, glutathione metabolism and nitrogen metabolism were enriched. By pathway enrichment and protein-protein interaction analyses, key DRPs such as glucose-6-phosphate 1-dehydrogenase, NADPH producing dehydrogenase, glutamate synthase and glutamine synthasewere identified.Additionally, salt responsive proteins of Jing724 were indicated to facilitate energy management, maintenance of redox homeostasis, reducing ammonia toxicity, osmotic homeostasis regulation, stress defense, stress adaptation, biotic cross-tolerance and gene transcription regulation. Quantification of multiple metabolic or enzymatic changes including SOD activity, MDA content, relative electrolyte leakage and Proline content were consistent with their predicted changes based on functions of DRPs. DRP analysis was correlated with the mRNA transcripts abundancevariation of eight representative DRPs. These results contribute to elucidating molecular networks of salt tolerance.

2017-12-05 | PXD007802 | Pride

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