An isoleucine residue acts as a thermal and regulatory switch in wheat Rubisco activase

Gustaf E. Degen; Dawn Worrall; Elizabete Carmo‐Silva

doi:10.1111/tpj.14766

The relative abundance of wheat Rubisco activase isoforms is post-transcriptionally regulated

Photosynthesis Research ◽

10.1007/s11120-021-00830-6 ◽

2021 ◽

Author(s):

Juan Alejandro Perdomo ◽

Peter Buchner ◽

Elizabete Carmo-Silva

Keyword(s):

Relative Abundance ◽

Light Availability ◽

Transcript Level ◽

Rubisco Activase ◽

Small Subunit ◽

Diel Cycle ◽

Diurnal Rhythms ◽

Crop Species ◽

Β Protein ◽

Wheat Leaves

AbstractDiurnal rhythms and light availability affect transcription–translation feedback loops that regulate the synthesis of photosynthetic proteins. The CO2-fixing enzyme Rubisco is the most abundant protein in the leaves of major crop species and its activity depends on interaction with the molecular chaperone Rubisco activase (Rca). In Triticum aestivum L. (wheat), three Rca isoforms are present that differ in their regulatory properties. Here, we tested the hypothesis that the relative abundance of the redox-sensitive and redox-insensitive Rca isoforms could be differentially regulated throughout light–dark diel cycle in wheat. While TaRca1-β expression was consistently negligible throughout the day, transcript levels of both TaRca2-β and TaRca2-α were higher and increased at the start of the day, with peak levels occurring at the middle of the photoperiod. Abundance of TaRca-β protein was maximal 1.5 h after the peak in TaRca2-β expression, but the abundance of TaRca-α remained constant during the entire photoperiod. The redox-sensitive TaRca-α isoform was less abundant, representing 85% of the redox-insensitive TaRca-β at the transcript level and 12.5% at the protein level. Expression of Rubisco large and small subunit genes did not show a consistent pattern throughout the diel cycle, but the abundance of Rubisco decreased by up to 20% during the dark period in fully expanded wheat leaves. These results, combined with a lack of correlation between transcript and protein abundance for both Rca isoforms and Rubisco throughout the entire diel cycle, suggest that the abundance of these photosynthetic enzymes is post-transcriptionally regulated.

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Ammonium application mitigates the effects of elevated carbon dioxide on the carbon/nitrogen balance of Phoebe bournei seedlings

Tree Physiology ◽

10.1093/treephys/tpab026 ◽

2021 ◽

Author(s):

Xiao Wang ◽

Xiaoli Wei ◽

Gaoyin Wu ◽

Shengqun Chen

Keyword(s):

Carbon Dioxide ◽

Enzyme Activities ◽

Atmospheric Carbon Dioxide ◽

Global Climate ◽

Co2 Concentration ◽

Rubisco Activase ◽

Plant Responses ◽

N Metabolism ◽

C And N ◽

Phoebe Bournei

Abstract The study of plant responses to increases in atmospheric carbon dioxide (CO2) concentration is crucial to understand and to predict the effect of future global climate change on plant adaptation and evolution. Increasing amount of nitrogen (N) can promote the positive effect of CO2, while how N forms would modify the degree of CO2 effect is rarely studied. The aim of this study was to determine whether the amount and form of nitrogen (N) could mitigate the effects of elevated CO2 (eCO2) on enzyme activities related to carbon (C) and N metabolism, the C/N ratio, and growth of Phoebe bournei (Hemsl.) Y.C. Yang. One-year-old P. bournei seedlings were grown in an open-top air chamber under either an ambient CO2 (aCO2) (350 ± 70 μmol•mol−1) or an eCO2 (700 ± 10 μmol•mol−1) concentration and cultivated in soil treated with either moderate (0.8 g per seedling) or high applications (1.2 g per seedling) of nitrate or ammonium. In seedlings treated with a moderate level of nitrate, the activities of key enzymes involved in C and N metabolism (i.e., Rubisco, Rubisco activase and glutamine synthetase) were lower under eCO2 than under aCO2. By contrast, key enzyme activities (except GS) in seedlings treated with high nitrate or ammonium were not significantly different between aCO2 and eCO2 or higher under eCO2 than under aCO2. The C/N ratio of seedlings treated with moderate or high nitrate under eCO2was significantly changed compared with the seedlings grown under aCO2, whereas the C/N ratio of seedlings treated with ammonium was not significantly different between aCO2 and eCO2. Therefore, under eCO2, application of ammonium can be beneficial C and N metabolism and mitigate effects on the C/N ratio.

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Proteome Analysis for Understanding Abiotic Stress (Salinity and Drought) Tolerance in Date Palm (Phoenix dactyliferaL.)

International Journal of Genomics ◽

10.1155/2015/407165 ◽

2015 ◽

Vol 2015 ◽

pp. 1-11 ◽

Cited By ~ 14

Author(s):

Haddad A. El Rabey ◽

Abdulrahman L. Al-Malki ◽

Khalid O. Abulnaja ◽

Wolfgang Rohde

Keyword(s):

Drought Stress ◽

Chlorophyll A ◽

Date Palm ◽

Light Harvesting ◽

Binding Protein ◽

Proteome Analysis ◽

Rubisco Activase ◽

Stress Conditions ◽

Spectrometric Analysis ◽

Salt And Drought Stress

This study was carried out to study the proteome of date palm under salinity and drought stress conditions to possibly identify proteins involved in stress tolerance. For this purpose, three-month-old seedlings of date palm cultivar “Sagie” were subjected to drought (27.5 g/L polyethylene glycol 6000) and salinity stress conditions (16 g/L NaCl) for one month. DIGE analysis of protein extracts identified 47 differentially expressed proteins in leaves of salt- and drought-treated palm seedlings. Mass spectrometric analysis identified 12 proteins; three out of them were significantly changed under both salt and drought stress, while the other nine were significantly changed only in salt-stressed plants. The levels of ATP synthase alpha and beta subunits, an unknown protein and some of RubisCO fragments were significantly changed under both salt and drought stress conditions. Changes in abundance of superoxide dismutase, chlorophyll A-B binding protein, light-harvesting complex1 protein Lhca1, RubisCO activase, phosphoglycerate kinase, chloroplast light-harvesting chlorophyll a/b-binding protein, phosphoribulokinase, transketolase, RubisCO, and some of RubisCO fragments were significant only for salt stress.

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Proteome Analysis of Date Palm (Phoenix dactylifera L.) under Severe Drought and Salt Stress

International Journal of Genomics ◽

10.1155/2016/7840759 ◽

2016 ◽

Vol 2016 ◽

pp. 1-8 ◽

Cited By ~ 6

Author(s):

Haddad A. El Rabey ◽

Abdulrahman L. Al-Malki ◽

Khalid O. Abulnaja

Keyword(s):

Salt Stress ◽

Drought Stress ◽

Drought Tolerance ◽

Date Palm ◽

Proteome Analysis ◽

Rubisco Activase ◽

2D Electrophoresis ◽

Severe Drought ◽

Protein Abundance ◽

Low Protein

Date palm cultivars differently tolerate salinity and drought stress. This study was carried out to study the response of date palm to severe salinity and drought based on leaf proteome analysis. Eighteen-month-old date palm plants were subjected to severe salt (48 g/L NaCl) and drought (82.5 g/L PEG or no irrigation) conditions for one month. Using a protein 2D electrophoresis method, 55 protein spots were analyzed using mass spectrometry. ATP synthase CF1 alpha chains were significantly upregulated under all three stress conditions. Changes in the abundance of RubisCO activase and one of the RubisCO fragments were significant in the same spots only for salt stress and drought stress with no irrigation, and oxygen-evolving enhancer protein 2 was changed in different spots. Transketolase was significantly changed only in drought stress with PEG. The expression of salt and drought stress genes of the chosen protein spots was either overexpressed or downexpressed as revealed by the high or low protein abundance, respectively. In addition, all drought tolerance genes due to no irrigation were downregulated. In conclusion, the proteome analysis of date palm under salinity and drought conditions indicated that both salinity and drought tolerance genes were differentially expressed resulting in high or low protein abundance of the chosen protein spots as a result of exposure to drought and salinity stress condition.

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Incorporation of a charged amino acid into the membrane-spanning domain blocks cell surface transport but not membrane anchoring of a viral glycoprotein

Molecular and Cellular Biology ◽

10.1128/mcb.5.6.1442-1448.1985 ◽

1985 ◽

Vol 5 (6) ◽

pp. 1442-1448

Author(s):

G A Adams ◽

J K Rose

Keyword(s):

Amino Acid ◽

Cell Surface ◽

G Protein ◽

Abnormal Behavior ◽

Animal Cells ◽

Viral Glycoprotein ◽

Protein Levels ◽

Membrane Spanning ◽

Membrane Anchoring ◽

Isoleucine Residue

The membrane-spanning domain of the vesicular stomatitis virus glycoprotein (G protein) consists of a continuous stretch of 20 uncharged and mostly hydrophobic amino acids. We examined the effects of two mutations which change the amino acid sequence in this domain. These mutations were generated by oligonucleotide-directed mutagenesis of a cDNA clone encoding the G protein, and the altered G proteins were then expressed in animal cells. Replacement of an isoleucine residue in the center of this domain with a strongly polar but uncharged amino acid (glutamine) had no effect on membrane anchoring or transport of the protein to the cell surface. Replacement of this same isoleucine residue with a charged amino acid (arginine) generated a G protein that still spanned intracellular membranes but was not transported efficiently to the cell surface. The protein accumulated in the Golgi region in about 50% of the cells, and about 20% of the cells had detectable protein levels in a punctate pattern on the cell surface. In the remaining cells the protein accumulated in a vesicular pattern throughout the cytoplasm. Models which might explain the abnormal behavior of this protein are discussed.

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Determining the Rate-Limiting Step for Light-Responsive Redox Regulation in Chloroplasts

Antioxidants ◽

10.3390/antiox7110153 ◽

2018 ◽

Vol 7 (11) ◽

pp. 153 ◽

Cited By ~ 6

Author(s):

Keisuke Yoshida ◽

Toru Hisabori

Keyword(s):

Redox Regulation ◽

Reducing Power ◽

Rubisco Activase ◽

Power Transfer ◽

Target Proteins ◽

Rate Limiting Step ◽

Limiting Step ◽

Rate Limiting

Thiol-based redox regulation ensures light-responsive control of chloroplast functions. Light-derived signal is transferred in the form of reducing power from the photosynthetic electron transport chain to several redox-sensitive target proteins. Two types of protein, ferredoxin-thioredoxin reductase (FTR) and thioredoxin (Trx), are well recognized as the mediators of reducing power. However, it remains unclear which step in a series of redox-relay reactions is the critical bottleneck for determining the rate of target protein reduction. To address this, the redox behaviors of FTR, Trx, and target proteins were extensively characterized in vitro and in vivo. The FTR/Trx redox cascade was reconstituted in vitro using recombinant proteins from Arabidopsis. On the basis of this assay, we found that the FTR catalytic subunit and f-type Trx are rapidly reduced after the drive of reducing power transfer, irrespective of the presence or absence of their downstream target proteins. By contrast, three target proteins, fructose 1,6-bisphosphatase (FBPase), sedoheptulose 1,7-bisphosphatase (SBPase), and Rubisco activase (RCA) showed different reduction patterns; in particular, SBPase was reduced at a low rate. The in vivo study using Arabidopsis plants showed that the Trx family is commonly and rapidly reduced upon high light irradiation, whereas FBPase, SBPase, and RCA are differentially and slowly reduced. Both of these biochemical and physiological findings suggest that reducing power transfer from Trx to its target proteins is a rate-limiting step for chloroplast redox regulation, conferring distinct light-responsive redox behaviors on each of the targets.

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