Frontiers of Sulfur Metabolism in Plant Growth, Development, and Stress Response

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07 April 2015
ATP-sulfurylase, sulfur-compounds, and plant stress tolerance
Naser A. Anjum
6 more and 
Sarvajeet S. Gill
Schematic representation of pathway of sulfate assimilation, reaction catalyzed by ATP-sulfurylase (ATP-S), and its regulation by major factors. Role of ATP-S in plant stress tolerance through sulfur/cysteine rich and sulfated compounds is outlined. Positive and negative regulation of ATP-S is indicated by arrows and blunt ends, respectively, [1Kawashima et al. (2011); 2Yatusevich et al. (2010); 3Hopkins et al. (2004); 4Schiavon et al. (2007); 5van de Mortel et al. (2008); 6Guo et al. (2009); 7Gill et al. (2012); 8Bashir et al. (2013); 9Asgher et al. (2014); 10Leao et al. (2014); 11Phartiyal et al. (2006); 12Ruiz and Blumwald (2002); 13Nazar et al. (2011); 14Passera et al. (1989); 15Huseby et al. (2013); 16Rotte and Leustek (2000); 17Takahashi et al. (1997); 18Liang et al. (2010); 19Lappartient and Touraine (1997); 20Lappartient and Touraine (1996); 21Vauclare et al. (2002)]. (APS, adenosine 5′-phosphosulfate; Cys, cysteine; AsA, ascorbate; GSH, reduced glutathione; PCs, phytochelatins; MTs, metallothioneins; ROS, reactive oxygen species).

Sulfur (S) stands fourth in the list of major plant nutrients after N, P, and K. Sulfate (SO42-), a form of soil-S taken up by plant roots is metabolically inert. As the first committed step of S-assimilation, ATP-sulfurylase (ATP-S) catalyzes SO42--activation and yields activated high-energy compound adenosine-5′-phosphosulfate that is reduced to sulfide (S2-) and incorporated into cysteine (Cys). In turn, Cys acts as a precursor or donor of reduced S for a range of S-compounds such as methionine (Met), glutathione (GSH), homo-GSH (h-GSH), and phytochelatins (PCs). Among S-compounds, GSH, h-GSH, and PCs are known for their involvement in plant tolerance to varied abiotic stresses, Cys is a major component of GSH, h-GSH, and PCs; whereas, several key stress-metabolites such as ethylene, are controlled by Met through its first metabolite S-adenosylmethionine. With the major aim of briefly highlighting S-compound-mediated role of ATP-S in plant stress tolerance, this paper: (a) overviews ATP-S structure/chemistry and occurrence, (b) appraises recent literature available on ATP-S roles and regulations, and underlying mechanisms in plant abiotic and biotic stress tolerance, (c) summarizes ATP-S-intrinsic regulation by major S-compounds, and (d) highlights major open-questions in the present context. Future research in the current direction can be devised based on the discussion outcomes.

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Sulfur is an essential macronutrient for plant growth and development. Reaching a thorough understanding of the molecular basis for changes in plant metabolism depending on the sulfur-nutritional status at the systems level will advance our basic knowledge and help target future crop improvement. Although the transcriptional responses induced by sulfate starvation have been studied in the past, knowledge of the regulation of sulfur metabolism is still fragmentary. This work focuses on the discovery of candidates for regulatory genes such as transcription factors (TFs) using ‘omics technologies. For this purpose a short term sulfate-starvation/re-supply approach was used. ATH1 microarray studies and metabolite determinations yielded 21 TFs which responded more than 2-fold at the transcriptional level to sulfate starvation. Categorization by response behaviors under sulfate-starvation/re-supply and other nutrient starvations such as nitrate and phosphate allowed determination of whether the TF genes are specific for or common between distinct mineral nutrient depletions. Extending this co-behavior analysis to the whole transcriptome data set enabled prediction of putative downstream genes. Additionally, combinations of transcriptome and metabolome data allowed identification of relationships between TFs and downstream responses, namely, expression changes in biosynthetic genes and subsequent metabolic responses. Effect chains on glucosinolate and polyamine biosynthesis are discussed in detail. The knowledge gained from this study provides a blueprint for an integrated analysis of transcriptomics and metabolomics and application for the identification of uncharacterized genes.

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Review
29 October 2014
Summary of regulatory mechanisms of response to sulfate starvation and resupply. Green text and arrows depict activation, while red text, arrows and lines represent repression. The interrupted lines symbolise the putative mechanisms of sensing of internal sulfate and transceptor function of SULTR1;2. CK abbreviates cytokinins, X represents xylem, and P phloem.
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Images show the typical distribution of glutathione. (A) Monochlorobimane staining in guard cells of the upper epidermis of tobacco cells in the light microscope. Fluorescence was observed in cytosol and nuclei (N) but not in vacuoles (V) and cell walls (arrowhead). Additionally, no fluorescence could be observed in chloroplasts (arrows in A and B) which can be best identified when comparing the autofluorescence of chloroplast (B) with monochlorobimane staining (A). Transmission electron micrographs show the subcellular distribution of glutathione (C,D) in mesophyll cells of leaves from Arabidopsis Col-0 plants. Glutathione-specific labeling could be observed in different concentrations in mitochondria (M), chloroplasts (C), peroxisomes (Px) but not in vacuoles (V) and cell walls (CW). Glutathione-specific labeling was observed in the stroma as well as inside the thylakoid lumen (arrowheads) when plants were exposed to high light intensities of 700 µmol m-2 s-1. Bars in (A,B) = 10 µm, (C,D) = 0.5 µm.
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Conserved regions I to IV of plant SOTs. Regions are shown as boxes, with size and position in the protein relative to the average size of SOTs from Arabidopsis thaliana. Functional amino acids were obtained from structural analyses of mouse SOTs as described above. The PAPS binding regions (5′P-motif, 3′P-motif, GxxGxxK), the proton acceptor histidine (His), and a poly-glutamic acid (Poly-Glu), that can be found in many plant SOTs, are labeled. The position of the Pfam domain PF00685 is shaded in gray.
Review
16 October 2014

All members of the sulfotransferase (SOT, EC 2.8.2.-) protein family transfer a sulfuryl group from the donor 3′-phosphoadenosine 5′-phosphosulfate (PAPS) to an appropriate hydroxyl group of several classes of substrates. The primary structure of these enzymes is characterized by a histidine residue in the active site, defined PAPS binding sites and a longer SOT domain. Proteins with this SOT domain occur in all organisms from all three domains, usually as a multi-protein family. Arabidopsis thaliana SOTs, the best characterized SOT multi-protein family, contains 21 members. The substrates for several plant enzymes have already been identified, such as glucosinolates, brassinosteroids, jasmonates, flavonoids, and salicylic acid. Much information has been gathered on desulfo-glucosinolate (dsGl) SOTs in A. thaliana. The three cytosolic dsGl SOTs show slightly different expression patterns. The recombinant proteins reveal differences in their affinity to indolic and aliphatic dsGls. Also the respective recombinant dsGl SOTs from different A. thaliana ecotypes differ in their kinetic properties. However, determinants of substrate specificity and the exact reaction mechanism still need to be clarified. Probably, the three-dimensional structures of more plant proteins need to be solved to analyze the mode of action and the responsible amino acids for substrate binding. In addition to A. thaliana, more plant species from several families need to be investigated to fully elucidate the diversity of sulfated molecules and the way of biosynthesis catalyzed by SOT enzymes.

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