NatB impacts growth and the response to osmotic stress in Arabidopsis
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PROVIDER: PRJNA773398 | ENA |
REPOSITORIES: ENA
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Project description:N-terminal acetylation (NTA) is one of the most abundant protein modifications in eukaryotes and is catalyzed in humans by seven Nα-acetyltransferases (NatA-F and NatH). Remarkably, the characterization of the plant Nat machinery and its biological relevance is still in its infancy, although NTA has gained recognition as key regulator of crucial processes like protein turnover, protein-protein interaction and protein targeting. In this study we combined in vitro assays, reverse genetics, quantitative N-terminomics, transcriptomics and physiological assays to characterize the Arabidopsis NatB complex. We show that the plant NatB catalytic (NAA20) and auxiliary subunit (NAA25) form a stable heterodimeric complex that accepts canonical NatB-type substrates in vitro. In planta, NatB complex formation was essential for enzymatic activity. Depletion of NatB subunits to 30% of wild-type level in three Arabidopsis T-DNA insertion mutants (naa20-1, naa20-2, naa25-1) decreased growth to 50% of wild-type level. A complementation approach revealed functional conservation between plant and human catalytic NatB subunits, while yeast NAA20 failed to complement naa20-1. Quantitative N-terminomics of approximately 2000 peptides identified 29 bona fide substrates of the plant NatB. In vivo, NatB preferentially acetylated N-termini starting with the initiator methionine followed by acidic amino acids and contributed 20% of the acetylation marks in the detected plant proteome. The global transcriptome and proteome analyses of NatB-depleted mutants suggested a function of NatB in multiple stress responses. In agreement, we revealed the specific impact of NatB on the resistance of plants to osmotic or high-salt stress. Remarkably, depletion of NatA did not affect these resistances.
2020-04-14 | GSE132978 | GEO
Project description:Downregulation of N-terminal initiator methionine imprinting by NatB impacts growth and the response to osmotic stress in Arabidopsis
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Project description:Arabidopsis Col-0 seeds were germinated and grown for two weeks on Arabidopsis thaliana salt media (ATS, control) or ATS media supplemented 50, 75, 100 or 125 mM NaCl that imposes both an ionic and osmotic stress; or ATS media supplemented with iso-osmolar concentrations of sorbitol (100, 150, 200 or 250 mM) that imposes only an osmotic stress. The aim of the study was to identify genes involved in plant growth and adaptation to ionic stress compared to genes involved in growth and adaptation to osmotic stress conditions. To do this we identified lists of genes that are differentially expressed in plants grown in NaCl (A) and lists of genes differentially expressed in plants grown in sorbitol (B). We then compared these lists to find ionic/salt-specific genes that are only expressed in plants grown in NaCl and not in plants grown in sorbitol; and osmotic genes that are expressed both in plants grown in NaCl and in plants grown in sorbitol. Associated publication: Cackett et al. (2022) Salt-specific gene expression reveals elevated auxin levels in Arabidopsis thaliana plants grown under saline conditions, DOI: 10.3389/fpls.2022.804716
2022-01-16 | GSE193762 | GEO
Project description:Genome-wide transcriptome analysis of Arabidopsis thaliana was performed to understand the role of auxin in the response of leaf growth to osmotic stress. We studied transcriptional changes in proliferating leaves of the seedlings grown in vitro on control medium, medium supplemented with 25mM mannitol, 0.1μM NAA and 0.1μM NAA + 25mM mannitol.
2021-05-09 | GSE60870 | GEO
Project description:Arabidopsis germination and two week growth on ionic vs osmotic stress
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Project description:determine the wide transcriptionnal response of Mtb to osmotic stress and hyperosmotic stress
2019-04-01 | GSE90839 | GEO
Project description:Drought is an important environmental factor affecting plant growth and biomass production. Despite this importance, little is known on the molecular mechanisms regulating plant growth under water limiting conditions. The main goal of this work was to investigate, using a combination of growth and molecular profiling techniques, how Arabidopsis thaliana leaves adapt their growth to prolonged mild osmotic stress. Fully proliferating, expanding and mature leaves were harvested from plants grown on plates without (control) or with 25mM mannitol (osmotic stress) and compared to seedlings at stage 1.03.
2010-04-11 | GSE16474 | GEO
Project description:Drought is an important environmental factor affecting plant growth and biomass production. Despite this importance little is known on the molecular mechanisms regulating plant growth under water limiting conditions. The main goal of this work was to investigate, using a combination of growth and molecular profiling techniques, how stress arrests CELl proliferation in Arabidopsis thaliana leaves upon osmotic stress imposition.
2011-07-01 | GSE22107 | GEO
Project description:Elucidating how plants sense and respond to water loss is important for identifying genetic and chemical interventions that may help sustain crop yields in water-limiting environments. Currently, the molecular mechanisms involved in the initial perception and response to dehydration are not well understood. Modern mass spectrometric methods for quantifying changes in the phosphoproteome provide an opportunity to identify key phosphorylation events involved in this process. Here, we have used both untargeted and targeted isotope-assisted mass spectrometric methods of phosphopeptide quantitation to characterize proteins in Arabidopsis (Arabidopsis thaliana) whose degree of phosphorylation is rapidly altered by hyperosmotic treatment. Thus, protein phosphorylation events responsive to 5 min of 0.3 m mannitol treatment were first identified using 15N metabolic labeling and untargeted mass spectrometry with a high-resolution ion-trap instrument. The results from these discovery experiments were then validated using targeted Selected Reaction Monitoring mass spectrometry with a triple quadrupole. Targeted Selected Reaction Monitoring experiments were conducted with plants treated under nine different environmental perturbations to determine whether the phosphorylation changes were specific for osmosignaling or involved cross talk with other signaling pathways. The results indicate that regulatory proteins such as members of the mitogen-activated protein kinase family are specifically phosphorylated in response to osmotic stress. Proteins involved in 5′ messenger RNA decapping and phosphatidylinositol 3,5-bisphosphate synthesis were also identified as targets of dehydration-induced phosphoregulation. The results of these experiments demonstrate the utility of targeted phosphoproteomic analysis in understanding protein regulation networks and provide new insight into cellular processes involved in the osmotic stress response.
2019-12-19 | PXD001057 | Pride
Project description:The euryhaline marine yeast Debaromyces hansenii is a model system for the study of genes related to osmotic stress. To study the transcriptional response of this organism to osmotic stress we have used the two color microarray based gene expression analysis of 6211 genes which constitutes the whole genome. Analysis was done at three time points after induction with salt (0.5h, 3h and 6h.) The mRNA level of 64 genes significantly increased at least 3-fold after induction with 2M NaCl whereas that of 45 genes was 3-fold diminished. The induced as well as the repressed genes were grouped into functional categories to identify biochemical processes possibly affected by osmotic shock. 53% of the induced genes encode for ribosomal proteins, 12.5% for mitochondrial and redox, 6% for aminoacid, cell wall and carbohydrate, and 1.5%for heat shock, protein transport and glycerol proteins. The function of 31% of repressed genes is currently unknown. 15% of the repressed genes are involved in carbohydrate metabolism and protective function, 6% are associated with cell wall, redox and signal transduction, and 4% in vacuolar and lipid functions. Surprisingly, the activity of NAD+ glycerol 3-phosphate dehydrogenase gene (GPD1) involved in the glycerol biosynthesis in response to osmotic stress did not show induction. Keywords: time course, stress response, mRNA expression
2008-03-15 | GSE9911 | GEO