Proteome Analysis of the ROF-FKBP Mutants Reveals Functional Relations among Heat Stress Responses, Plant Development, and Protein Quality Control during Heat Acclimation in Arabidopsis thaliana
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SUBMITTER: Lefa P
PROVIDER: S-EPMC10795062 | biostudies-literature | 2023 Dec
REPOSITORIES: biostudies-literature
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Project description:In the present study, a differential screening following heat stress acclimation was performed in A. thaliana WT and ROF-FKBP mutated plants, using mass spectrometry, and the results were used to understand and analyze the effect of the ROF PPIases during thermotolerance acquisition in plants. Our data highlight the central role of these two PPIases in heat stress and point to their direct or indirect effect on other proteins participating in cellular functions such as protein folding and quality control, cell division, photosynthesis, and other metabolic and signaling processes. 1) Specifically, the heat stress response, protein folding and protein ER processing pathways are enhanced following a 37 oC acclimation period independently to the mutation state. 2) However, at 37 oC and in the double mutated rof1–/2 –plants, a higher accumulation of proteins belonging to the above pathways is observed compared to all other conditions (WT, mutants, control and heat stressed). 3) Furthermore, the proteasomal pathway including the common member of both the protasomal and lysosomal degradation pathway CDC48, is over-represented in the extracts of both untreated and heat stressed rof1–/2 – mutants compared to the other extracts. 4) In contrast, in the single rof1 – mutation the heat acclimation (including ROF2), protein folding and proteasomal pathways are significantly suppressed compared to the WT, at 37 oC. 5) Protein accumulation related to the heat stress and protein quality control pathways follow the rof1–/2 – > rof2 – > rof1 – pattern suggesting a differential but also synergistic role of the two proteins. 6) Protein complexes of other biochemical pathways such as the Lhc of the photosynthetic pathway and the phosphoinositide binding proteins involved in membrane-trafficking events during cell plate formation and cytokinesis (Patellin-1, 2 and 3) are negatively regulated by the rof1–/2 – mutants. Our results suggest that ROF1 and ROF2 FKBPs regulate stress response, developmental and metabolic pathways via a complex feedback mechanism that ensures the continuation of protein quality control at their absence.
2024-01-26 | PXD044216 | Pride
Project description:Plants are subjected to strong fluctuations in light intensity in their natural growth environment, caused both by unpredictable changes due to weather conditions and movement of clouds and upper canopy leaves and predictable changes during day-night cycle. The mechanisms of long-term acclimation to fluctuating light (FL) are still not well understood. Here, we used quantitative mass spectrometry to investigate long-term acclimation of low light-grown Arabidopsis thaliana to a FL condition that induces mild photooxidative stress. On the third day of exposure to FL, young and mature leaves were harvested in the morning and at the end of day for proteome analysis using a stable isotope labeling approach. We identified 2,313 proteins, out of which 559 proteins exhibited significant changes in abundance in at least one of the four experimental groups (morning-young, morning-mature, end-of-day-young, end-of-day-mature). A core set of 49 proteins showed significant responses to FL in three or four experimental groups, which included enhanced accumulation of proteins involved in photoprotection, cyclic electron flow around photosystem I, photorespiration, and glycolysis, while specific glutathione transferases and proteins involved in translation and chlorophyll biosynthesis were reduced in abundance. In addition, we observed pathway- and protein-specific changes predominantly at the end of day, whereas few changes were observed exclusively in the morning. Comparison of the proteome data with the matching transcript data revealed gene- and protein-specific responses, with several chloroplast-localized proteins decreasing in abundance despite increased gene expression under FL. Together, our data shows moderate but widespread alterations of protein abundance during acclimation to FL and suggests an important role of post-transcriptional regulation of protein abundance.
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Project description:Plants can obtain superinduction of defense against unpredictable challenges based on prior acclimation, but the mechanisms involved in the acclimation memory are little known. The objective of this study was to characterize mechanisms of heat acclimation memory in Rhododendron hainanense, a thermotolerant wild species of azalea. Pretreatment of a 2-d recovery (25/18°C, day/night) after heat acclimation (37°C, 1 h) (AR-pt) did not weaken but enhanced acquired thermotolerance in R. hainanense with less damaged phenotype, net photosynthetic rate, and membrane stability than non-acclimation pretreated (NA-pt) plants. Combined transcriptome and proteome analysis revealed that a lot of heat-responsive genes still maintained high protein abundance rather than transcript level after the 2-d recovery. Photosynthesis-related genes were highly enriched and most decreased under heat stress (HS: 42°C, 1 h) with a less degree in AR-pt plants compared to NA-pt. Sustainably accumulated chloroplast-localized heat shock proteins (HSPs), Rubisco activase 1 (RCA1), beta-subunit of chaperonin-60 (CPN60β), and plastid transcriptionally active chromosome 5 (pTAC5) in the recovery period probably provided equipped protection of AR-pt plants against the subsequent HS, with less damaged photochemical efficiency and chloroplast structure. In addition, significant higher levels of RCA1 transcripts in AR-pt compared to NA-pt plants in early stage of HS showed a more important role of RCA1 than other chaperonins in heat acclimation memory. The novel heat-induced RCA1, rather than constitutively expressed RCA2 and RCA3, showed excellent thermostability after long-term HS (LHS: 42/35°C, 7 d) and maintained balanced Rubisco activation state in photosynthetic acclimation. This study provides new insights into plant heat acclimation memory and indicates candidate genes for genetic modification and molecular breeding in thermotolerance improvement.
| S-EPMC7466565 | biostudies-literature
Project description:A mutant of Arabidopsis thaliana (L.) Heyn. (a small plant in the crucifer family) that lacks glycine decarboxylase activity owing to a recessive nuclear mutation has been isolated on the basis of a growth requirement for high concentrations of atmospheric CO2. Mitochondria isolated from leaves of the mutant did not exhibit glycine-dependent O2 consumption, did not release 14CO2 from [14C]glycine, and did not catalyse the glycine-bicarbonate exchange reaction that is considered to be the first partial reaction associated with glycine cleavage. Photosynthesis in the mutant was decreased after illumination under atmospheric conditions that promote partitioning of carbon into intermediates of the photorespiratory pathway, but was not impaired under non-photorespiratory conditions. Thus glycine decarboxylase activity is not required for any essential function unrelated to photorespiration. The photosynthetic response of the mutant in photorespiratory conditions is probably caused by an increased rate of glyoxylate oxidation, which results from the sequestering of all readily transferable amino groups in a metabolically inactive glycine pool, and by a depletion of intermediates from the photosynthesis cycle. The rate of release of 14CO2 from exogenously applied [14C]glycollate was 14-fold lower in the mutant than in the wild type, suggesting that glycine decarboxylation is the only significant source of photorespiratory CO2.
| S-EPMC1158121 | biostudies-other
Project description:BackgroundGlobal warming is a major challenge for plant survival and growth. Understanding the molecular mechanisms by which higher plants sense and adapt to upsurges in the ambient temperature is essential for developing strategies to enhance plant tolerance to heat stress. Here, we designed a heat-responsive Arabidopsis thaliana reporter line that allows an in-depth investigation of the mechanisms underlying the accumulation of protective heat-shock proteins (HSPs) in response to high temperature.MethodsA transgenic Arabidopsis thaliana reporter line named "Heat-Inducible Bioluminescence And Toxicity" (HIBAT) was designed to express from a conditional heat-inducible promoter, a fusion gene encoding for nanoluciferase and D-amino acid oxidase, whose expression is toxic in the presence of D-valine. HIBAT seedlings were exposed to different heat treatments in presence or absence of D-valine and analyzed for survival rate, bioluminescence and HSP gene expression.ResultsWhereas at 22 °C, HIBAT seedlings grew unaffected by D-valine, and all survived iterative heat treatments without D-valine, 98% died following heat treatments on D-valine. The HSP17.3B promoter was highly specific to heat as it remained unresponsive to various plant hormones, Flagellin, H2O2, osmotic stress and high salt. RNAseq analysis of heat-treated HIBAT seedlings showed a strong correlation with expression profiles of two wild type lines, confirming that HIBAT does not significantly differ from its Col-0 parent. Using HIBAT, a forward genetic screen revealed candidate loss-of-function mutants, apparently defective either at accumulating HSPs at high temperature or at repressing HSP accumulation at non-heat-shock temperatures.ConclusionHIBAT is a valuable candidate tool to identify Arabidopsis mutants defective in the response to high temperature stress. It opens new avenues for future research on the regulation of HSP expression and for understanding the mechanisms of plant acquired thermotolerance.
| S-EPMC10251684 | biostudies-literature
Project description:Leaves of Arabidopsis thaliana transferred from low to high light increase their capacity for photosynthesis, a process of dynamic acclimation. A mutant, gpt2, lacking a chloroplast glucose-6-phosphate/phosphate translocator, is deficient in its ability to acclimate to increased light. Here, we have used a label-free proteomics approach, to perform relative quantitation of 1993 proteins from Arabidopsis wild type and gpt2 leaves exposed to increased light. Data are available via ProteomeXchange with identifier PXD006598. Acclimation to light is shown to involve increases in electron transport and carbon metabolism but no change in the abundance of photosynthetic reaction centers. The gpt2 mutant shows a similar increase in total protein content to wild type but differences in the extent of change of certain proteins, including in the relative abundance of the cytochrome b6f complex and plastocyanin, the thylakoid ATPase and selected Benson-Calvin cycle enzymes. Changes in leaf metabolite content as plants acclimate can be explained by changes in the abundance of enzymes involved in metabolism, which were reduced in gpt2 in some cases. Plants of gpt2 invest more in stress-related proteins, suggesting that their reduced ability to acclimate photosynthetic capacity results in increased stress.
| S-EPMC5517461 | biostudies-literature
Project description:In plants, environmental perturbations often result in oxidative reactions in the apoplastic space, which are counteracted for by enzymatic and non-enzymatic antioxidative systems, including ascorbate and glutathione. However, the occurrence of the latter and its exact role in the extracellular space are not well documented. In Arabidopsis thaliana, the gamma-glutamyl transferase isoform GGT1 bound to the cell wall takes part in the so-called gamma-glutamyl cycle for extracellular glutathione degradation and recovery, and may be implicated in redox sensing and balance. In this work, oxidative conditions were imposed with UV-B radiation and studied in redox altered ggt1 mutants. Elevated UV-B has detrimental effects on plant metabolism, plasma membranes representing a major target for ROS generated by this harmful radiation. The response of ggt1 knockout Arabidopsis leaves to UV-B radiation was assessed by investigating changes in apoplastic protein composition. We then compared the expression changes resulting from the mutation and from the UV-B treatment. Rearrangements occurring in apoplastic protein composition suggest the involvement of hydrogen peroxide, which may ultimately act as a signal. Other important changes related to hormonal effects, cell wall remodeling, and redox activities are also reported. We argue that oxidative stress conditions imposed by UV-B and by disruption of the gamma-glutamyl cycle result in similar stress-induced responses, to some degree at least. Data shown here are associated with the article from Trentin et al. (2015) [1]; protein data have been deposited to the PRIDE database (Vizcaíno et al., 2014) [2] with identifier PXD001807.
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Project description:Phosphite (H₂PO⁻₃) induces a range of physiological and developmental responses in plants by disturbing the homeostasis of the macronutrient phosphate. Because of its close structural resemblance to phosphate, phosphite impairs the sensing, membrane transport, and subcellular compartmentation of phosphate. In addition, phosphite induces plant defence responses by an as yet unknown mode of action. In this study, the acclimation of Arabidopsis thaliana plants to a sustained phosphite supply in the growth medium was investigated and compared with plants growing under varying phosphate supplies. Unlike phosphate, phosphite did not suppress the formation of lateral roots in several Arabidopsis accessions. In addition, the expression of well-documented phosphate-starvation-induced genes, such as miRNA399d and At4, was not repressed by phosphite accumulation, whilst the induction of PHT1;1 and PAP1 was accentuated. Thus, a mimicking of phosphate by phosphite was not observed for these classical phosphate-starvation responses. Metabolomic analysis of phosphite-treated plants showed changes in several metabolite pools, most prominently those of aspartate, asparagine, glutamate, and serine. These alterations in amino acid pools provide novel insights for the understanding of phosphite-induced pathogen resistance.
| S-EPMC3617837 | biostudies-literature
Project description:Cold acclimation (CA) leads to increased plant freezing tolerance during exposure to low, non-freezing temperatures as a result of many physiological, biochemical and molecular changes that have been extensively investigated. In addition, many plant species, such as Arabidopsis thaliana, respond to a subsequent exposure to mild, non-damaging freezing temperatures with an additional increase in freezing tolerance referred to as sub-zero acclimation (SZA). There is comparatively little information available about the molecular basis of SZA. However, previous transcriptomic studies indicated that cell wall modification may play an important role during SZA. Here we show that CA and SZA are accompanied by extensive changes in cell wall amount, composition and structure. While CA leads to a significant increase in cell wall amount, the relative proportions of pectin, hemicellulose and cellulose remained unaltered during both CA and SZA. However, both treatments resulted in more subtle changes in structure as determined by infrared spectroscopy and monosaccharide composition as determined by gas chromatography-mass spectrometry. These differences could be related through a proteomic approach to the accumulation of cell wall modifying enzymes such as pectin methylesterases, pectin methylesterase inhibitors and xyloglucan endotransglucosylases/hydrolases in the extracellular matrix.
| S-EPMC6381082 | biostudies-literature
Project description:BackgroundCold stress causes dynamic changes in gene expression that are partially caused by small non-coding RNAs since they regulate protein coding transcripts and act in epigenetic gene silencing pathways. Thus, a detailed analysis of transcriptional changes of small RNAs (sRNAs) belonging to all known sRNA classes such as microRNAs (miRNA) and small interfering RNA (siRNAs) in response to cold contributes to an understanding of cold-related transcriptome changes.ResultWe subjected A. thaliana plants to cold acclimation conditions (4 °C) and analyzed the sRNA transcriptomes after 3 h, 6 h and 2 d. We found 93 cold responsive differentially expressed miRNAs and only 14 of these were previously shown to be cold responsive. We performed miRNA target prediction for all differentially expressed miRNAs and a GO analysis revealed the overrepresentation of miRNA-targeted transcripts that code for proteins acting in transcriptional regulation. We also identified a large number of differentially expressed cis- and trans-nat-siRNAs, as well as sRNAs that are derived from long non-coding RNAs. By combining the results of sRNA and mRNA profiling with miRNA target predictions and publicly available information on transcription factors, we reconstructed a cold-specific, miRNA and transcription factor dependent gene regulatory network. We verified the validity of links in the network by testing its ability to predict target gene expression under cold acclimation.ConclusionIn A. thaliana, miRNAs and sRNAs derived from cis- and trans-NAT gene pairs and sRNAs derived from lncRNAs play an important role in regulating gene expression in cold acclimation conditions. This study provides a fundamental database to deepen our knowledge and understanding of regulatory networks in cold acclimation.
| S-EPMC7325139 | biostudies-literature