Ethylene modulates the dynamics of translation via GCN2 and EIN2 in Arabidopsis under submergence
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PROVIDER: PRJNA628644 | ENA |
REPOSITORIES: ENA
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Project description:General translational repression is predicted as a key process to reduce energy consumption under hypoxia. We have previously showed that mRNA loading onto polysome is reduced in Arabidopsis under submergence. Here, we showed that plant stress activated GCN2 (general control nonderepressible 2) can phosphorylate eIF2a (Eukaryotic Initiation Factor 2a) in Arabidopsis under submergence, and this process is reversible after desubmergence. Compared to the wild-type, the reduction in polysome loading during submergence was less severe in the gcn2 mutant. Transgenic lines overexpressing GCN2 had more ATP and conferred better tolerance under submergence, suggesting that GCN2 might modulate the dynamics of translation to adjust the energy homeostasis under hypoxia. Interestingly, GCN2-eIF2a signaling was activated by ethylene under submergence. However, GCN2 activity was not affected in ein2-5 and eil1ein3 under submergence, suggesting that GCN2 activity was regulated by noncanonical ethylene signaling. In addition, the polysome loading was retained in both ein2-5 and etr1-1 under submergence, implying that ethylene modulated the dynamic translation under submergence via EIN2 and GCN2. Notably, our NGS analysis also demonstrated that EIN2 and GCN2 regulated the translation of 23 core hypoxia genes as well as 53% translational repressed genes under submergence. On the other hand, EIN2 and GCN2 also affected the expression of genes involved in hypoxic response, ethylene response, biotic stress and negative regulation of cytokinin signaling. Taken together, these demonstrated that entrapped ethylene triggers GCN2 and EIN2 to ensure the translation of stress required proteins under submergence and also provide a step stone for future investigation how eukaryotic cells modulate the translation to response for the changeable environments.
2022-06-08 | GSE149414 | GEO
Project description:A complex interplay between ethylene, ETP1/ETP2 F-box proteins, and degradation of EIN2 is essential for triggering ethylene responses in plants. The gaseous plant hormone ethylene can trigger myriad physiological and morphological responses in plants. While many ethylene signaling pathway components have been identified and characterized, little is known about the function of the integral membrane protein EIN2, a central regulator of all ethylene responses. Here, we demonstrate that Arabidopsis thaliana EIN2 is a protein with a short half-life that undergoes rapid proteasome-mediated protein turnover. Moreover, EIN2 protein accumulation is positively regulated by ethylene. We identified two F-box proteins, EIN2 TARGETING PROTEIN and 2 (ETP1 and ETP2), that interact with the EIN2 carboxyl-terminal domain (CEND), which is highly conserved and sufficient to activate most ethylene responses. Overexpression of ETP1 or ETP2 disrupts EIN2 protein accumulation, and these plants manifest a strong ethylene insensitive phenotype. Furthermore, knocking down the levels of both ETP1 and ETP2 mRNAs using an artificial microRNA (amiRNA) leads to accumulation of EIN2 protein, resulting in plants that display constitutive ethylene response phenotypes. Finally, ethylene down-regulates ETP1 and ETP2 proteins, impairing their ability to interact with EIN2. Thus, these studies reveal that a complex interplay between ethylene, the regulation of ETP1/ETP2 F-box proteins, and subsequent targeting and degradation of EIN2 is essential for triggering ethylene responses in plants. Keywords: ethylene treatment, genetic modification
2009-12-01 | GSE14247 | GEO
Project description:A complex interplay between ethylene, ETP1/ETP2 F-box proteins, and degradation of EIN2 is essential for triggering ethylene responses in plants. The gaseous plant hormone ethylene can trigger myriad physiological and morphological responses in plants. While many ethylene signaling pathway components have been identified and characterized, little is known about the function of the integral membrane protein EIN2, a central regulator of all ethylene responses. Here, we demonstrate that Arabidopsis thaliana EIN2 is a protein with a short half-life that undergoes rapid proteasome-mediated protein turnover. Moreover, EIN2 protein accumulation is positively regulated by ethylene. We identified two F-box proteins, EIN2 TARGETING PROTEIN and 2 (ETP1 and ETP2), that interact with the EIN2 carboxyl-terminal domain (CEND), which is highly conserved and sufficient to activate most ethylene responses. Overexpression of ETP1 or ETP2 disrupts EIN2 protein accumulation, and these plants manifest a strong ethylene insensitive phenotype. Furthermore, knocking down the levels of both ETP1 and ETP2 mRNAs using an artificial microRNA (amiRNA) leads to accumulation of EIN2 protein, resulting in plants that display constitutive ethylene response phenotypes. Finally, ethylene down-regulates ETP1 and ETP2 proteins, impairing their ability to interact with EIN2. Thus, these studies reveal that a complex interplay between ethylene, the regulation of ETP1/ETP2 F-box proteins, and subsequent targeting and degradation of EIN2 is essential for triggering ethylene responses in plants. Experiment Overall Design: Six samples were analyzed. There were three treatments with two biological replicates each. The treatments are as follows: 8 week old Col-0 plants (air control), 8 week old Col-0 plants treated for 24 hours with ethylene gas (10 ppm), and artificial microRNA knockdown mutants, amiR-ETP1/2.
2009-12-05 | E-GEOD-14247 | biostudies-arrayexpress
Project description:Seed size is related to plant evolution and crop yield and is affected by genetic mutations, imprinting, and genome dosage. Imprinting is a widespread epigenetic phenomenon in mammals and flowering plants. ETHYLENE INSENSITIVE2 (EIN2) encodes a membrane protein that links the ethylene perception to transcriptional regulation. Interestingly, during seed development EIN2 is maternally-expressed in Arabidopsis and maize, but the role of EIN2 in seed development is unknown. Here we show that EIN2 is expressed specifically in the endosperm, and the maternal-specific EIN2 expression affects temporal regulation of endosperm cellularization. As a result, seed size increases in the genetic cross using the ein2 mutant as the maternal parent or in the ein2 mutant. The maternal-specific expression of EIN2 in the endosperm is controlled by DNA methylation but not by H3K27me3 or by ethylene and several ethylene pathway genes tested. RNA-seq analysis in the endosperm isolated by laser-capture microdissection show upregulation of many endosperm-expressed genes such as AGAMOUS-LIKEs (AGLs) in the ein2 mutant or when the maternal EIN2 allele is not expressed. EIN2 does not interact with DNA and may act through ETHYLENE INSENSITIVE3 (EIN3), a DNA binding protein present in sporophytic tissues, to activate target genes like AGLs, which in turn mediate temporal regulation of endosperm cellularization and seed size. These results provide mechanistic insights into endosperm and maternal-specific expression of EIN2 on endosperm cellularization and seed development, which could help improve seed production in plants and crops.
2022-10-12 | GSE147587 | GEO
Project description:We report that ethylene regulate the ENAP1 binding. We perform a dual cross-linking procedure using the additional cross-linker ethylene glycol bis-succinimidylsuccinate (EGS; Thermo Scientific) along with formaldehyde according to (Pedmale et al. 2016) for Chip-sequencing (ChIPseq) of ENAP1, as well as ENAP1 ChIP- EIN2 reChIPseq (ChIP-reChIPseq), which were used chromatins isolated from 3-day old etiolated Col-0 seedlings treated with ethylene or air gas. Results show that ENAP1 is involved in the EIN2 dependent ethylene response.
2017-09-07 | GSE97288 | GEO
Project description:Ethylene gas is essential for many developmental processes and stress responses in plants. ETHYLENE INSENSITIVE2 (EIN2), an NRAMP-homologous integral membrane protein, plays an essential role in ethylene signaling but its function remains enigmatic. Here we report that phosphorylation-regulated proteolytic processing of EIN2 triggers its endoplasmic reticulum (ER)-nucleus translocation, which is essential for hormone signaling and response in Arabidopsis. Without ethylene, or in hormone receptors mutants, ER-tethered EIN2 shows CTR1 kinase-dependent phosphorylation. Ethylene exposure triggers dephosphorylation and proteolytic cleavage, resulting in rapid nuclear translocation of a carboxyl-terminal EIN2 fragment (C’). Plants containing mutations that mimic EIN2 dephosphorylation, or inactivate CTR1, show constitutive cleavage and nuclear localization of EIN2-C’, and EIN3/EIL1-dependent activation of ethylene responses. These findings uncover a mechanism of subcellular communication whereby ethylene gas stimulates rapid phosphorylation-dependent cleavage and nuclear movement of the EIN2-C’ peptide, thus linking hormone perception and signaling components located in the ER with nuclear-localized transcriptional regulators.
2012-10-22 | E-MTAB-4360 | biostudies-arrayexpress
Project description:The Arabidopsis mutant cir1 displayed constitutive expression of defence genes and enhanced resistance to the virulent pathogens Pseudomonas syringae pv tomato (Pst) and Peronospora parasitica Noco2. In order to dissect the downstream signalling components constitutively activated in cir1, cir1 plants were crossed to the ethylene-insensitive mutant ein2. Resistance to Pst was abolished in cir1:ein2 plantswhile resistance to P. parasitica Noco2 was maintained. Thus CIR1-mediated resistance to Pst appears to be dependent on ethylene signalling through EIN2but resistance to P. parasitica Noco2 is EIN2-independent. The aim of the proposed experiment is to identify EIN2 dependent and independent global gene expression in cir1 plants. It is likely that expression of a sub-set of EIN2-dependent genes in cir1 are required for resistance to Pst whereas EIN2-independent genes may be required for P.parasitica Noco2 resistance. Identification and analysis of genes in these sub-sets may reveal common regulatory mechanisms and may extend the understanding of resistance to Pst and P. parasitica Noco2 in Arabidopsis. This information should be of interest to the Arabidopsis disease resistance community. Total RNA will be extracted from leaf tissue harvested from five-week old soil-grown plants.
2003-10-30 | E-NASC-7 | biostudies-arrayexpress
Project description:Understanding how developmental and environmental signals are integrated to produce specific responses is one of the main challenges of modern biology. Hormones and, most importantly, interactions between different hormones serve as crucial regulators of plant growth and development, playing central roles in the coordination of internal developmental processes with the environment. Herein, a combination of physiological, genetic, cellular, and whole-genome expression profiling approaches has been employed to investigate the mechanisms of interaction between two key plant hormones, ethylene and auxin. Quantification of the morphological effects of ethylene and auxin in a variety of mutant backgrounds indicates that auxin biosynthesis, transport, signaling and response are required for the ethylene-induced growth inhibition in roots but not in hypocotyls. Analysis of the activation of early auxin and ethylene responses at cellular level, as well as of global changes in gene expression in the wild type versus auxin and ethylene mutants, suggests a simple mechanistic model for the interaction between these two hormones in roots. This model not only implies existence of several levels of interaction but also provides a likely explanation for the strong ethylene response defects observed in auxin mutants. Experiment Overall Design: The effect of ethylene on gene expression in Col-0 (wild-type) and aux1-7 (an auxin resistant mutant) seedlings' roots was investigated. Hydrocarbon-free air was used as control for the ethylene treatment. Two biological replicates were examined. Experiment Overall Design: Similarly, the effect of auxin (IAA) on gene expression in Col (wild-type) and ein2-5 (an ethylene insensitive mutant) seedlings' roots was studied. Two biological replicates were employed.
2008-06-15 | E-GEOD-7432 | biostudies-arrayexpress
Project description:We report that ethylene reduce the ENAP1 binding. We perform Chip-sequencing of ENAP1 using chromatins isolated from 3-day old etiolated Col-0/ein2/ein3eil1 seedlings treated with ethylene or air gas. Results show that ethylene reduce ENAP1 binding in TSS region of EIN3 targeted genes that are regulated at the transcriptional level by ethylene.
2017-09-07 | GSE81200 | GEO
Project description:We reported that ethylene specifically elevated acetylation of histone H3 at K14 and the non-canonical acetylation of histone H3 at K23,but not H3K9ac.Thus, we further performed Chip-sequencing of H3K9Ac, using chromatins isolated from 3-day old etiolated ein2-5 seedlings treated with ethylene or air gas.
2017-08-11 | GSE93875 | GEO