Publications (18)108.73 Total impact
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Article: Regulation of flowering by trehalose-6-phosphate signaling in Arabidopsis thaliana.
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ABSTRACT: The timing of the induction of flowering determines to a large extent the reproductive success of plants. Plants integrate diverse environmental and endogenous signals to ensure the timely transition from vegetative growth to flowering. Carbohydrates are thought to play a crucial role in the regulation of flowering, and trehalose-6-phosphate (T6P) has been suggested to function as a proxy for carbohydrate status in plants. The loss of TREHALOSE-6-PHOSPHATE SYNTHASE 1 (TPS1) causes Arabidopsis thaliana to flower extremely late, even under otherwise inductive environmental conditions. This suggests that TPS1 is required for the timely initiation of flowering. We show that the T6P pathway affects flowering both in the leaves and at the shoot meristem, and integrate TPS1 into the existing genetic framework of flowering-time control.Science 02/2013; 339(6120):704-7. · 31.20 Impact Factor -
Article: RobiNA: a user-friendly, integrated software solution for RNA-Seq-based transcriptomics.
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ABSTRACT: Recent rapid advances in next generation RNA sequencing (RNA-Seq)-based provide researchers with unprecedentedly large data sets and open new perspectives in transcriptomics. Furthermore, RNA-Seq-based transcript profiling can be applied to non-model and newly discovered organisms because it does not require a predefined measuring platform (like e.g. microarrays). However, these novel technologies pose new challenges: the raw data need to be rigorously quality checked and filtered prior to analysis, and proper statistical methods have to be applied to extract biologically relevant information. Given the sheer volume of data, this is no trivial task and requires a combination of considerable technical resources along with bioinformatics expertise. To aid the individual researcher, we have developed RobiNA as an integrated solution that consolidates all steps of RNA-Seq-based differential gene-expression analysis in one user-friendly cross-platform application featuring a rich graphical user interface. RobiNA accepts raw FastQ files, SAM/BAM alignment files and counts tables as input. It supports quality checking, flexible filtering and statistical analysis of differential gene expression based on state-of-the art biostatistical methods developed in the R/Bioconductor projects. In-line help and a step-by-step manual guide users through the analysis. Installer packages for Mac OS X, Windows and Linux are available under the LGPL licence from http://mapman.gabipd.org/web/guest/robin.Nucleic Acids Research 06/2012; 40(Web Server issue):W622-7. · 8.03 Impact Factor -
Article: Mutagenesis of cysteine 81 prevents dimerization of the APS1 subunit of ADP‐glucose pyrophosphorylase and alters diurnal starch turnover in Arabidopsis thaliana leaves
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ABSTRACT: Many plants, including Arabidopsis thaliana, retain a substantial portion of their photosynthate in leaves in the form of starch, which is remobilized to support metabolism and growth at night. ADP-glucose pyrophosphorylase (AGPase) catalyses the first committed step in the pathway of starch synthesis, the production of ADP-glucose. The enzyme is redox-activated in the light and in response to sucrose accumulation, via reversible breakage of an intermolecular cysteine bridge between the two small (APS1) subunits. The biological function of this regulatory mechanism was investigated by complementing an aps1 null mutant (adg1) with a series of constructs containing a full-length APS1 gene encoding either the wild-type APS1 protein or mutated forms in which one of the five cysteine residues was replaced by serine. Substitution of Cys81 by serine prevented APS1 dimerization, whereas mutation of the other cysteines had no effect. Thus, Cys81 is both necessary and sufficient for dimerization of APS1. Compared to control plants, the adg1/APS1C81S lines had higher levels of ADP-glucose and maltose, and either increased rates of starch synthesis or a starch-excess phenotype, depending on the daylength. APS1 protein levels were five- to tenfold lower in adg1/APS1C81S lines than in control plants. These results show that redox modulation of AGPase contributes to the diurnal regulation of starch turnover, with inappropriate regulation of the enzyme having an unexpected impact on starch breakdown, and that Cys81 may play an important role in the regulation of AGPase turnover.The Plant Journal 12/2011; 70(2):231 - 242. · 6.16 Impact Factor -
Article: Use of TILLING and robotised enzyme assays to generate an allelic series of Arabidopsis thaliana mutants with altered ADP-glucose pyrophosphorylase activity.
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ABSTRACT: ADP-glucose pyrophosphorylase (AGPase) catalyses the synthesis of ADP-glucose, and is a highly regulated enzyme in the pathway of starch synthesis. In Arabidopsis thaliana, the enzyme is a heterotetramer, containing two small subunits encoded by the APS1 gene and two large subunits encoded by the APL1-4 genes. TILLING (Targeting Induced Local Lesions IN Genomes) of a chemically mutagenised population of A. thaliana plants identified 33 novel mutations in the APS1 gene, including 21 missense mutations in the protein coding region. High throughput measurements using a robotised cycling assay showed that maximal AGPase activity in the aps1 mutants varied from <15 to 117% of wild type (WT), and that the kinetic properties of the enzyme were altered in several lines, indicating a role for the substituted amino acid residues in catalysis or substrate binding. These results validate the concept of using such a platform for efficient high-throughput screening of very large populations of mutants, natural accessions or introgression lines. AGPase was estimated to have a flux control coefficient of 0.20, indicating that the enzyme exerted only modest control over the rate of starch synthesis in plants grown under short day conditions (8 h light/16 h dark) with an irradiance of 150 μmol quanta m(-2)s(-1). Redox activation of the enzyme, via reduction of the intermolecular disulphide bridge between the two small subunits, was increased in several lines. This was sometimes, but not always, associated with a decrease in the abundance of the APS1 protein. In conclusion, the TILLING technique was used to generate an allelic series of aps1 mutants in A. thaliana that revealed new insights into the multi-layered regulation of AGPase. These mutants offer some advantages over the available loss-of-function mutants, e.g. adg1, for investigating the effects of subtle changes in the enzyme's activity on the rate of starch synthesis.Journal of plant physiology 02/2011; 168(12):1395-405. · 2.50 Impact Factor -
Article: AtTPS1-mediated trehalose 6-phosphate synthesis is essential for embryogenic and vegetative growth and responsiveness to ABA in germinating seeds and stomatal guard cells.
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ABSTRACT: Trehalose and associated metabolites are part of the sugar signalling system in plants and have profound effects on development. Disruption of the TREHALOSE 6-PHOSPHATE SYNTHASE (TPS1) gene in Arabidopsis results in delayed embryo growth, altered cell wall morphology and carbon metabolism and abortion at the torpedo stage. Here we investigate the role of the TPS1 gene in post-embryonic development using two approaches. In the first we use the seed-specific ABI3 promoter to drive the TPS1 cDNA during embryo development, resulting in rescue of the embryo-lethal tps1 phenotype. Lack of expression from the ABI3::TPS1 transgene in post-germinative tps1 seedlings results in severe growth arrest, accumulation of soluble sugars and starch and leads to an increase in expression of genes related to ABA signalling. In the second approach we use TILLING (targeted induced local lesions in genomes) to generate three weaker, non-embryo-lethal, alleles (tps1-11, tps1-12 and tps1-13) and use these to demonstrate that the TPS1 protein plays a key role in modulating trehalose 6-phosphate (T6P) levels in vegetative tissues of Arabidopsis. All three weaker alleles give a consistent phenotype of slow growth and delayed flowering. Germination of tps1-11, tps1-12 and tps1-13 is hypersensitive to ABA with the degree of hypersensitivity correlating with the decrease in T6P levels in the different alleles. Stomatal pore aperture is regulated by ABA, and this was found to be affected in tps1-12. Our results show that the TPS1 gene product plays an essential role in regulating the growth of vegetative as well as embryogenic tissue in a mechanism involving ABA and sugar metabolism.The Plant Journal 10/2010; 64(1):1-13. · 6.16 Impact Factor -
Article: Sucrose non-fermenting kinase 1 (SnRK1) coordinates metabolic and hormonal signals during pea cotyledon growth and differentiation.
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ABSTRACT: Seed development passes through developmental phases such as cell division, differentiation and maturation: each have specific metabolic demands. The ubiquitous sucrose non-fermenting-like kinase (SnRK1) coordinates and adjusts physiological and metabolic demands with growth. In protoplast assays sucrose deprivation and hormone supplementation, such as with auxin and abscisic acid (ABA), stimulate SnRK1-promoter activity. This indicates regulation by nutrients: hormonal crosstalk under conditions of nutrient demand and cell proliferation. SnRK1-repressed pea (Pisum sativum) embryos show lower cytokinin levels and deregulation of cotyledonary establishment and growth, together with downregulated gene expression related to cell proliferation, meristem maintenance and differentiation, leaf formation, and polarity. This suggests that at early stages of seed development SnRK1 regulates coordinated cotyledon emergence and growth via cytokinin-mediated auxin transport and/or distribution. Decreased ABA levels and reduced gene expression, involved in ABA-mediated seed maturation and response to sugars, indicate that SnRK1 is required for ABA synthesis and/or signal transduction at an early stage. Metabolic profiling of SnRK1-repressed embryos revealed lower levels of most organic and amino acids. In contrast, levels of sugars and glycolytic intermediates were higher or unchanged, indicating decreased carbon partitioning into subsequent pathways such as the tricarbonic acid cycle and amino acid biosynthesis. It is hypothesized that SnRK1 mediates the responses to sugar signals required for early cotyledon establishment and patterning. As a result, later maturation and storage activity are strongly impaired. Changes observed in SnRK1-repressed pea seeds provide a framework for how SnRK1 communicates nutrient and hormonal signals from auxins, cytokinins and ABA to control metabolism and development.The Plant Journal 10/2009; 61(2):324-38. · 6.16 Impact Factor -
Article: Adjustment of growth, starch turnover, protein content and central metabolism to a decrease of the carbon supply when Arabidopsis is grown in very short photoperiods.
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ABSTRACT: Arabidopsis was grown in a 12, 8, 4 or 3 h photoperiod to investigate how metabolism and growth adjust to a decreased carbon supply. There was a progressive increase in the rate of starch synthesis, decrease in the rate of starch degradation, decrease of malate and fumarate, decrease of the protein content and decrease of the relative growth rate. Carbohydrate and amino acids levels at the end of the night did not change. Activities of enzymes involved in photosynthesis, starch and sucrose synthesis and inorganic nitrogen assimilation remained high, whereas five of eight enzymes from glycolysis and organic acid metabolism showed a significant decrease of activity on a protein basis. Glutamate dehydrogenase activity increased. In a 2 h photoperiod, the total protein content and most enzyme activities decreased strongly, starch synthesis was inhibited, and sugars and amino acids levels rose at the end of the night and growth was completely inhibited. The rate of starch degradation correlated with the protein content and the relative growth rate across all the photoperiod treatments. It is discussed how a close coordination of starch turnover, the protein content and growth allows Arabidopsis to avoid carbon starvation, even in very short photoperiods.Plant Cell and Environment 03/2009; 32(7):859-74. · 5.22 Impact Factor -
Chapter: Sucrose Metabolism
12/2008; , ISBN: 9780470015902 -
Article: Decreased expression of cytosolic pyruvate kinase in potato tubers leads to a decline in pyruvate resulting in an in vivo repression of the alternative oxidase.
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ABSTRACT: The aim of this work was to investigate the effect of decreased cytosolic pyruvate kinase (PKc) on potato (Solanum tuberosum) tuber metabolism. Transgenic potato plants with strongly reduced levels of PKc were generated by RNA interference gene silencing under the control of a tuber-specific promoter. Metabolite profiling showed that decreased PKc activity led to a decrease in the levels of pyruvate and some other organic acids involved in the tricarboxylic acid cycle. Flux analysis showed that this was accompanied by changes in carbon partitioning, with carbon flux being diverted from glycolysis toward starch synthesis. However, this metabolic shift was relatively small and hence did not result in enhanced starch levels in the tubers. Although total respiration rates and the ATP to ADP ratio were largely unchanged, transgenic tubers showed a strong decrease in the levels of alternative oxidase (AOX) protein and a corresponding decrease in the capacity of the alternative pathway of respiration. External feeding of pyruvate to tuber tissue or isolated mitochondria resulted in activation of the AOX pathway, both in the wild type and the PKc transgenic lines, providing direct evidence for the regulation of AOX by changes in pyruvate levels. Overall, these results provide evidence for a crucial role of PKc in the regulation of pyruvate levels as well as the level of the AOX in heterotrophic plant tissue, and furthermore reveal that these parameters are interlinked in vivo.Plant physiology 11/2008; 148(3):1640-54. · 6.53 Impact Factor -
Article: Compartmentation in plant metabolism.
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ABSTRACT: Cell fractionation and immunohistochemical studies in the last 40 years have revealed the extensive compartmentation of plant metabolism. In recent years, new protein mass spectrometry and fluorescent-protein tagging technologies have accelerated the flow of information, especially for Arabidopsis thaliana, but the intracellular locations of the majority of proteins in the plant proteome are still not known. Prediction programs that search for targeting information within protein sequences can be applied to whole proteomes, but predictions from different programs often do not agree with each other or, indeed, with experimentally determined results. The compartmentation of most pathways of primary metabolism is generally covered in plant physiology textbooks, so the focus here is mainly on newly discovered metabolic pathways in plants or pathways that have recently been revised. Ultimately, all of the pathways of plant metabolism are interconnected, and a major challenge facing plant biochemists is to understand the regulation and control of metabolic networks. One of the best-characterized networks links sucrose synthesis in the cytosol with photosynthetic CO(2) fixation and starch synthesis in the chloroplasts. One of the key features of this network is how the transport of pathway intermediates and signal metabolites across the chloroplast envelope conveys information between the two compartments, influencing the regulation of several enzymes to co-ordinate fluxes through the different pathways. It is widely accepted that chloroplasts and mitochondria originated from prokaryotic endosymbionts, and that new transporters and regulatory networks evolved to integrate metabolism in these organelles with the rest of the cell. Curiously, the present-day locations of many metabolic pathways within the cell often do not reflect their evolutionary origin, and there is evidence of extensive shuffling of enzymes and whole pathways between compartments during the evolution of plants.Journal of Experimental Botany 02/2007; 58(1):35-47. · 5.36 Impact Factor -
Article: Sugar-induced increases in trehalose 6-phosphate are correlated with redox activation of ADPglucose pyrophosphorylase and higher rates of starch synthesis in Arabidopsis thaliana.
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ABSTRACT: Tre6P (trehalose 6-phosphate) is implicated in sugar-signalling pathways in plants, but its exact functions in vivo are uncertain. One of the main obstacles to discovering these functions is the difficulty of measuring the amount of Tre6P in plant tissues. We have developed a highly specific assay, using liquid chromatography coupled to MS-Q3 (triple quadrupole MS), to measure Tre6P in the femto-picomole range. The Tre6P content of sucrose-starved Arabidopsis thaliana seedlings in axenic culture increased from 18 to 482 pmol x g(-1) FW (fresh weight) after adding sucrose. Leaves from soil-grown plants contained 67 pmol x g(-1) FW at the end of the night, which rose to 108 pmol x g(-1)FW after 4 h of illumination. Even greater changes in Tre6P content were seen after a 6 h extension of the dark period, and in the starchless mutant, pgm. The intracellular concentration of Tre6P in wild-type leaves was estimated to range from 1 to 15 microM. It has recently been reported that the addition of Tre6P to isolated chloroplasts leads to redox activation of AGPase (ADPglucose pyrophosphorylase) [Kolbe, Tiessen, Schluepmann, Paul, Ulrich and Geigenberger (2005) Proc. Natl. Acad. Sci. U.S.A. 102, 11118-11123]. Using the new assay for Tre6P, we found that rising sugar levels in plants are accompanied by increases in the level of Tre6P, redox activation of AGPase and the stimulation of starch synthesis in vivo. These results indicate that Tre6P acts as a signalling metabolite of sugar status in plants, and support the proposal that Tre6P mediates sucrose-induced changes in the rate of starch synthesis.Biochemical Journal 08/2006; 397(1):139-48. · 4.90 Impact Factor -
Article: Evolution and function of the sucrose-phosphate synthase gene families in wheat and other grasses.
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ABSTRACT: Suc-phosphate synthase (SPS) is a key regulatory enzyme in the pathway of Suc biosynthesis and has been linked to quantitative trait loci controlling plant growth and yield. In dicotyledonous plants there are three SPS gene families: A, B, and C. Here we report the finding of five families of SPS genes in wheat (Triticum aestivum) and other monocotyledonous plants from the family Poaceae (grasses). Three of these form separate subfamilies within the previously described A, B, and C gene families, but the other two form a novel and distinctive D family, which on present evidence is only found in the Poaceae. The D-type SPS proteins lack the phosphorylation sites associated with 14-3-3 protein binding and osmotic stress activation, and the linker region between the N-terminal catalytic glucosyltransferase domain and the C-terminal Suc-phosphatase-like domain is 80 to 90 amino acid residues shorter than in the A, B, or C types. The D family appears to have arisen after the divergence of mono- and dicotyledonous plants, with a later duplication event resulting in the two D-type subfamilies. Each of the SPS gene families in wheat showed different, but overlapping, spatial and temporal expression patterns, and in most organs at least two different SPS genes are expressed. Analysis of expressed sequence tags indicated similar expression patterns to wheat for each SPS gene family in barley (Hordeum vulgare) but not in more distantly related grasses. We identified an expressed sequence tag from rice (Oryza sativa) that appears to be derived from an endogenous antisense SPS gene, and this might account for the apparently low level of expression of the related OsSPS11 sense gene, adding to the already extensive list of mechanisms for regulating the activity of SPS in plants.Plant physiology 08/2004; 135(3):1753-64. · 6.53 Impact Factor -
Article: New complexities in the synthesis of sucrose.
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ABSTRACT: Sucrose is universal in plants and fulfils many roles: transport sugar, storage reserve, compatible solute and signal compound. Consequently, sucrose synthesis is highly regulated, with much of the control operating at the first step in the committed pathway, which is catalysed by sucrose-phosphate synthase (SPS). The discovery of at least three SPS gene families in plants has added a further layer of complexity to an already complicated picture involving transcriptional, allosteric and post-translational control of this enzyme's activity. After years of neglect, the gene encoding the last enzyme in the pathway, sucrose-phosphatase (SPP), has finally been cloned, revealing that SPS contains an SPP-like domain at the carboxy-terminus, to which SPP might bind. This has reinvigorated the search for an SPS-SPP complex, and has hinted at further complexities to be unravelled in the control of sucrose synthesis in plants.Current Opinion in Plant Biology 07/2003; 6(3):208-14. · 9.27 Impact Factor -
Article: Expression of a cyanobacterial sucrose-phosphate synthase from Synechocystis sp. PCC 6803 in transgenic plants.
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ABSTRACT: Sucrose-phosphate synthase (SPS) from the cyanobacterium Synechocystis sp. PCC 6803 lacks all of the Ser residues known to be involved in the regulation of higher plant SPS by protein phosphorylation. The Synechocystis SPS is also not allosterically regulated by glucose 6-phosphate or orthophosphate. To investigate the effects of expressing a potentially unregulated SPS in plants, the Synechocystis sps gene was introduced into tobacco, rice and tomato under the control of constitutive promoters. The Synechocystis SPS protein was expressed at a high level in the plants, which should have been sufficient to increase overall SPS activity 2-8-fold in the leaves. However, SPS activities and carbon partitioning in leaves from transgenic and wild-type plants were not significantly different. The maximal light-saturated rates of photosynthesis in leaves from tomato plants expressing the Synechocystis SPS were the same as those from wild-type plants. Tomato plants expressing the maize SPS showed 2-3-fold increases in SPS activity, increased partitioning of photoassimilate to sucrose and up to 58% higher maximal rates of photosynthesis. To investigate the apparent inactivity of the Synechocystis SPS the enzyme was purified from transgenic tobacco and rice plants. Surprisingly, the purified enzyme was found to have full catalytic activity. It is proposed that some other protein in plant cells binds to the Synechocystis SPS resulting in inhibition of the enzyme.Journal of Experimental Botany 02/2003; 54(381):223-37. · 5.36 Impact Factor -
Article: Sucrose-phosphatase gene families in plants.
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ABSTRACT: Sucrose-phosphatase (SPP; EC 3.1.3.24) catalyzes the final step in the pathway of sucrose biosynthesis and higher plants contain multiple isoforms of the enzyme encoded by different genes. The genome of the dicotyledonous plant Arabidopsis thaliana (thale cress) contains four SPP-like genes on chromosomes 1 (AtSPP1), 2 (AtSPP2) and 3 (AtSPP3a and AtSPP3b), all of which are expressed. The genome of the monocotyledonous plant rice (Oryza sativa) also contains four SPP-like genes, which have very similar exon-intron structures to those from A. thaliana. Two cDNA clones that encode catalytically active SPP enzymes have been isolated from maize (Zea mays), showing that this species contains at least two functional SPP genes. Multiple SPP-like cDNA clones have also been identified from wheat (Triticum aestivum), barley (Hordeum vulgare) and tomato (Lycopersicon esculentum). The genomes of two cyanobacteria, Synechocystis sp. PCC 6803 and Anabaena sp. PCC 7120, contain single spp genes. The cyanobacterial SPPs and the N-terminal region of the higher plant enzyme share significant similarity with members of the haloacid dehalogenase (HAD) superfamily of hydrolases/phosphatases. In addition to the HAD phosphatase domain, SPP from higher plants also contains a shorter, C-terminal domain of unknown function. An SPP-like sequence from the bryophyte (moss) Physcomitrella patens also contains this C-terminal domain, indicating that its acquisition was an early event in the evolution of higher plants.Gene 02/2003; 303:187-96. · 2.34 Impact Factor -
Article: Adenosine 5′-triphosphate-mediated activation of sucrose-phosphate synthase in bundle sheath cells of C4 plants
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ABSTRACT: We report the ATP-mediated activation of sucrose-phosphate synthase in bundle sheath cells prepared from C4 species. Sucrose synthesis was followed by measuring the incorporation of [14C]fructose 6-phosphate into sucrose in bundle sheath cells also provided with uridine 5′-diphosphoglucose (UDPGlc). Studies with Panicum miliaceum L. cells showed that activation was largely due to an increase in the affinity for UDPGlc and was therefore only evident at limiting UDPGlc concentrations. The apparent K m UDPGlc for sucrose synthesis by cells pretreated and assayed with ATP was about 0.7 mM compared with 7–8 mM for control cells without ATP. The γ-thio derivative of ATP had a similar effect to ATP. The effect was also evident when ATP was rapidly removed from cells prior to assay. Sucrose-phosphate synthase activity in extracts from cells pretreated with or without ATP showed similar differences in K m UDPGlc. These observations support the view that ATP is inducing a covalent modification of the enzyme. However, several protein kinase inhibitors did not prevent activation. Changes of more than threefold were observed for the K m UDPGlc with sucrose-phosphate synthase extracted from bundle sheath cells rapidly isolated from attached leaves that were subjected to dark/light treatments. The possible relationship between these changes and those induced by ATP with isolated cells is discussed.Planta 04/1997; 202(2):249-256. · 3.00 Impact Factor -
Article: Sugar-induced increases in trehalose 6-phosphate are correlated with redox activation of ADPglucose pyrophosphorylase and higher rates of starch synthesis in Arabidopsis thaliana
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ABSTRACT: Final full-text version of the paper available at: http://www.biochemj.org/ Tre6P (trehalose 6-phosphate) is implicated in sugar-signalling pathways in plants, but its exact functions in vivo are uncertain. One of the main obstacles to discovering these functions is the difficulty of measuring the amount of Tre6P in plant tissues. We have developed a highly specific assay, using liquid chromatography coupled to MS-Q3 (triple quadrupole MS), to measure Tre6P in the femto-picomole range. The Tre6P content of sucrose-starved Arabidopsis thaliana seedlings in axenic culture increased from 18 to 482 pmol·g−1FW (fresh weight) after adding sucrose. Leaves from soil-grown plants contained 67 pmol·g−1FW at the end of the night, which rose to 108 pmol·g−1FW after 4 h of illumination. Even greater changes in Tre6P content were seen after a 6 h extension of the dark period, and in the starchless mutant, pgm. The intracellular concentration of Tre6P in wild-type leaves was estimated to range from 1 to 15 μM. It has recently been reported that the addition of Tre6P to isolated chloroplasts leads to redox activation of AGPase (ADPglucose pyrophosphorylase) [Kolbe, Tiessen, Schluepmann, Paul, Ulrich and Geigenberger (2005) Proc. Natl. Acad. Sci. U.S.A. 102, 11118–11123]. Using the new assay for Tre6P, we found that rising sugar levels in plants are accompanied by increases in the level of Tre6P, redox activation of AGPase and the stimulation of starch synthesis in vivo. These results indicate that Tre6P acts as a signalling metabolite of sugar status in plants, and support the proposal that Tre6P mediates sucrose-induced changes in the rate of starch synthesis. This research was supported by theMax Planck Society and by the BMBF (Federal Ministry of Education and Research)-funded project GABI (Genomanalyse im biologischen System Pflanze) Verbund Arabidopsis III ‘Gauntlets, Carbon and Nutrient Signaling: Test Systems, and Metabolite and Transcript Profiles’ (0312277A). R.M. was supported by a Ramón y Cajal research contract. We thank Melanie Höhne for excellent technical assistance. We also thank Dr Michael Dahl (University of Konstanz, Germany) and Dr Carlos Gancedo (Unidad de Bioquimica y Genetica de Levaduras, Instituto de Investigaciones Biomedicas Alberto Sols, C. S. I. C.- Universidad Autonoma de Madrid, Madrid, Spain) for the gifts of the pSG2 plasmid and Yarrowia lipolytica HXK1 over-expressing yeast strain respectively. Peer reviewed -
Article: Purification and properties of sucrose-phosphate synthase from seeds of Pisum sativum
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ABSTRACT: The aim of this work was to purify and characterize sucrose-phosphate synthase from 38 hr germinated seeds of Pisum sativum. Chromatography on ω-aminohexyl Sepharose 4B and PD 10 Sephadex G-25M followed by fast protein liquid chromatography on a Mono Q anion exchange column and a Superose 6 gel filtration column gave a preparation of specific activity 4.22 μmol min−1 mg−1 protein. SDS PAGE showed four major and five minor bands. Native molecular mass was 456 000. Activity was optimum at pH 7.0 and 5 mM magnesium chloride. Hyperbolic kinetics were found for UDPglucose, apparent Km 2.4 mM and, in the presence of glucose 6-phosphate, for fructose 6-phosphate. No inhibition was detected with either sucrose or sucrose phosphate. Glucose 6-phosphate and fructose-1,6-bisphosphate stimulated activity, but Pi and UDP were inhibitory.Phytochemistry.
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- The Plant Journal (3)
- Journal of Experimental Botany (2)
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- Planta (1)
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Institutions
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2003–2011
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Max-Planck-Institut für molekulare Pflanzenphysiologie
Potsdam, Brandenburg, Germany -
Max-Planck-Gesellschaft
München, Bavaria, Germany
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2006
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Second University of Naples
Caserta, Campania, Italy
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