Characterization of two soluble inorganic pyrophosphatases from Arabidopsis thaliana

Sancha, Ernesto Navarro-De la; Coello-Coutiño, Martha P.; Valencia-Turcotte, Lilián Gabriela; Hernández-Domínguez, Eric E.; Trejo-Yepes, Gisela; Rodríguez‐Sotres, Rogelio

doi:10.1016/j.plantsci.2006.12.011

Cited by 24 publications

(25 citation statements)

References 35 publications

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“…60% and still remained at a very high level of nearly 5000 μmol Pi mg −1 min −1 , which is probably sufficient to inhibit pollen growth. MdPPa has a lower sPPase activity than p26.1; this may be attributed to different characteristics in different plant species (Gavalas & Manetas, ; Hara et al ., ; du Jardin et al ., ; Visser et al ., ; Navarro‐De la Sancha et al ., ). In this study, we found that the in vitro addition of S ‐RNases to the PPase assay shows a decrease in PPase activity to a maximum of 50%.…”

Section: Discussionmentioning

confidence: 97%

Apple S‐RNase triggers inhibition of tRNA aminoacylation by interacting with a soluble inorganic pyrophosphatase in growing self‐pollen tubes in vitro

Meng

et al. 2018

New Phytologist

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Apple exhibits S-RNase-based self-incompatibility (SI), in which S-RNase plays a central role in rejecting self-pollen. It has been proposed that the arrest of pollen growth in SI of Solanaceae plants is a consequence of the degradation of pollen rRNA by S-RNase; however, the underlying mechanism in Rosaceae is still unclear. Here, we used S -RNase as a bait to screen an apple pollen cDNA library and characterized an apple soluble inorganic pyrophosphatase (MdPPa) that physically interacted with S-RNases. When treated with self S-RNases, apple pollen tubes showed a marked growth inhibition, as well as a decrease in endogenous soluble pyrophosphatase activity and elevated levels of inorganic pyrophosphate (PPi). In addition, S-RNase was found to bind to two variable regions of MdPPa, resulting in a noncompetitive inhibition of its activity. Silencing of MdPPa expression led to a reduction in pollen tube growth. Interestingly, tRNA aminoacylation was inhibited in self S-RNase-treated or MdPPa-silenced pollen tubes, resulting in the accumulation of uncharged tRNA. Furthermore, we provide evidence showing that this disturbance of tRNA aminoacylation is independent of RNase activity. We propose an alternative mechanism differing from RNA degradation to explain the cytotoxicity of the S-RNase apple SI process.

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Section: Discussionmentioning

confidence: 97%

Apple S‐RNase triggers inhibition of tRNA aminoacylation by interacting with a soluble inorganic pyrophosphatase in growing self‐pollen tubes in vitro

Meng

et al. 2018

New Phytologist

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“…Cytosolic localization of the gene product has been confirmed by microscopic visualization of At1g01050-GFP fusion proteins in transgenic plants (Koroleva et al, 2005). Heterologous expression of PPA1 in Escherichia coli and in-depth characterization of enzyme activity demonstrate that this enzyme is a monomeric, Mg 2+ -dependent phosphatase that is strictly specific to the pyrophosphate substrate (Navarro-De la Sancha et al, 2007).…”

Section: Identification and Characterization Of A Dominant Low-seed-omentioning

confidence: 96%

“…While the characterization of the lo15571 event alone, in particular the observation of increased At1g01050 protein levels in developing siliques, already strongly supports a connection between altered PPiase expression and the low-seed-oil phenotype, it does not clarify to which extent the seed phenotype is caused by alterations in carbohydrate metabolism in nonseed tissues, including source and other sink organs of the plant. This is due to the fact that At1g01050 is expressed during most stages of plant development (Navarro-De la Sancha et al, 2007) and that we do not know if and how patterns of At1g01050 expression are altered at the whole plant level due to the enhancer tag insertion. However, analysis of the GY1::At1g01050 events clarifies this and shows that increased At1g01050 protein abundance during the maturation phase of developing seed, most likely embryo tissues, leads to reduced conversion of sugars to seed storage lipid.…”

Section: Gene Validation: Characterization Of Transgenic Events With mentioning

confidence: 99%

Oil and Protein Accumulation in Developing Seeds Is Influenced by the Expression of a Cytosolic Pyrophosphatase in Arabidopsis

et al. 2012

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This study describes a dominant low-seed-oil mutant (lo15571) of Arabidopsis (Arabidopsis thaliana) generated by enhancer tagging. Compositional analysis of developing siliques and mature seeds indicated reduced conversion of photoassimilates to oil. Immunoblot analysis revealed increased levels of At1g01050 protein in developing siliques of lo15571. At1g01050 encodes a soluble, cytosolic pyrophosphatase and is one of five closely related genes that share predicted cytosolic localization and at least 70% amino acid sequence identity. Expression of At1g01050 using a seed-preferred promoter recreated most features of the lo15571 seed phenotype, including low seed oil content and increased levels of transient starch and soluble sugars in developing siliques. Seed-preferred RNA interference-mediated silencing of At1g01050 and At3g53620, a second cytosolic pyrophosphatase gene that shows expression during seed filling, led to a heritable oil increase of 1% to 4%, mostly at the expense of seed storage protein. These results are consistent with a scenario in which the rate of mobilization of sucrose, for precursor supply of seed storage lipid biosynthesis by cytosolic glycolysis, is strongly influenced by the expression of endogenous pyrophosphatase enzymes. This emphasizes the central role of pyrophosphate-dependent reactions supporting cytosolic glycolysis during seed maturation when ATP supply is low, presumably due to hypoxic conditions. This route is the major route providing precursors for seed oil biosynthesis. ATP-dependent reactions at the entry point of glycolysis in the cytosol or plastid cannot fully compensate for the loss of oil content observed in transgenic events with increased expression of cytosolic pyrophosphatase enzyme in the cytosol. These findings shed new light on the dynamic properties of cytosolic pyrophosphate pools in developing seed and their influence on carbon partitioning during seed filling. Finally, our work uniquely demonstrates that genes encoding cytosolic pyrophosphatase enzymes provide novel targets to improve seed composition for plant biotechnology applications.

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“…In addition to orthologs of sPPase, plants also contain vacuolar H + -pumping inorganic pyrophosphatase (H + -PPase). Arabidopsis thaliana contains six paralogs of sPPase, PPa1 to PPa6 (Schulze et al, 2004;Navarro-De la Sancha et al, 2007). sPPase isozymes are classified into two families: Mg 2+dependent enzymes in Family I and Mn 2+ -dependent enzymes in Family II.…”

Section: Introductionmentioning

confidence: 99%

“…The subcellular localization of plant sPPases has been examined using several methods: immunogold labeling for potato (Solanum tuberosum) enzymes (Rojas-Beltrán et al, 1999), analysis of GFP fusion proteins for Arabidopsis enzymes (Koroleva et al, 2005;Öztürk et al, 2014;Gutiérrez-Luna et al, 2016), and an in vitro import assay for pea (Pisum sativum) PPa6 (to chloroplasts; Schulze et al, 2004). PPi hydrolysis activity has been detected for several sPPase isozymes in plants (Schulze et al, 2004;Gómez-García et al, 2006;Navarro-De la Sancha et al, 2007). Nonetheless, the physiological roles of these cytosolic sPPases are not fully understood.…”

Section: Introductionmentioning

confidence: 99%

Vacuolar H⁺-Pyrophosphatase and Cytosolic Soluble Pyrophosphatases Cooperatively Regulate Pyrophosphate Levels in Arabidopsis thaliana

et al. 2018

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Inorganic pyrophosphate (PPi) is a phosphate donor and energy source. Many metabolic reactions that generate PPi are suppressed by high levels of PPi. Here, we investigated how proper levels of cytosolic PPi are maintained, focusing on soluble pyrophosphatases (AtPPa1 to AtPPa5; hereafter PPa1 to PPa5) and vacuolar H-pyrophosphatase (H-PPase, AtVHP1/FUGU5) in In planta, five PPa isozymes tagged with GFP were detected in the cytosol and nuclei. Immunochemical analyses revealed a high abundance of PPa1 and the absence of PPa3 in vegetative tissue. In addition, the heterologous expression of each PPa restored growth in a soluble PPase-defective yeast strain. Although the quadruple knockout mutant plant showed no obvious phenotypes, H-PPase and PPa1 double mutants () exhibited significant phenotypes, including dwarfism, high PPi concentrations, ectopic starch accumulation, decreased cellulose and callose levels, and structural cell wall defects. Altered cell arrangements and weakened cell walls in the root tip were particularly evident in and were more severe than in Our results indicate that H-PPase is essential for maintaining adequate PPi levels and that the cytosolic PPa isozymes, particularly PPa1, prevent increases in PPi concentrations to toxic levels. We discuss phenotypes in relation to metabolic reactions and PPi homeostasis.

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Characterization of two soluble inorganic pyrophosphatases from Arabidopsis thaliana

Cited by 24 publications

References 35 publications

Apple S‐RNase triggers inhibition of tRNA aminoacylation by interacting with a soluble inorganic pyrophosphatase in growing self‐pollen tubes in vitro

Apple S‐RNase triggers inhibition of tRNA aminoacylation by interacting with a soluble inorganic pyrophosphatase in growing self‐pollen tubes in vitro

Oil and Protein Accumulation in Developing Seeds Is Influenced by the Expression of a Cytosolic Pyrophosphatase in Arabidopsis

Vacuolar H⁺-Pyrophosphatase and Cytosolic Soluble Pyrophosphatases Cooperatively Regulate Pyrophosphate Levels in Arabidopsis thaliana

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Characterization of two soluble inorganic pyrophosphatases from Arabidopsis thaliana

Cited by 24 publications

References 35 publications

Apple S‐RNase triggers inhibition of tRNA aminoacylation by interacting with a soluble inorganic pyrophosphatase in growing self‐pollen tubes in vitro

Apple S‐RNase triggers inhibition of tRNA aminoacylation by interacting with a soluble inorganic pyrophosphatase in growing self‐pollen tubes in vitro

Oil and Protein Accumulation in Developing Seeds Is Influenced by the Expression of a Cytosolic Pyrophosphatase in Arabidopsis

Vacuolar H+-Pyrophosphatase and Cytosolic Soluble Pyrophosphatases Cooperatively Regulate Pyrophosphate Levels in Arabidopsis thaliana

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Vacuolar H⁺-Pyrophosphatase and Cytosolic Soluble Pyrophosphatases Cooperatively Regulate Pyrophosphate Levels in Arabidopsis thaliana