Plastidic P2 glucose-6P dehydrogenase from poplar is modulated by thioredoxin m-type: Distinct roles of cysteine residues in redox regulation and NADPH inhibition

Cardi, Manuela; Zaffagnini, Mirko; Lillo, Alessia De; Castiglia, Daniela; Chibani, Kamel; Gualberto, José M.; Rouhier, Nicolas; Jacquot, Jean‐Pierre; Esposito, Sergio

doi:10.1016/j.plantsci.2016.08.003

Cited by 24 publications

(28 citation statements)

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“…It has been reported that the plant G6PDHs perform significant functions in maintaining the cellular redox balance (Krüger et al, 2011;Cardi et al, 2016). To validate whether overexpression of cytosolic GmG6PDH2 gene could affect the redox fluctuation, the redox states of AsA and GSH were monitored in soybean transgenic hair roots.…”

Section: Gmg6pdh2 Is Essential For Asa and Gsh Biosynthesis Under Salmentioning

confidence: 99%

“…Cytosolic (Cy) and plastidic (P) isoforms have been certified for plant G6PDHs based on their subcellular localization (Cardi et al, 2013;Castiglia et al, 2015). In addition, the P-G6PDHs are divided into two types, P1-G6PDH and P2-G6PDH, which can be distinguished by diverse gene expression profiles as well as specific biochemical characteristics, indicating the different functions of each isoform in plant metabolism (Cardi et al, 2016). In Arabidopsis, there are six G6PDHs targeted to different subcellular compartments: two cytosolic NADP + -dependent isoforms encoded by AtG6PDH5 and AtG6PDH6 genes respectively, and four plastidic NADP +dependent isoforms encoded by AtG6PDH1, AtG6PDH2, AtG6PDH3, and AtG6PDH4 genes respectively (Wakao and Benning, 2005).…”

Section: Introductionmentioning

confidence: 99%

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Genome-Wide Analysis of the Glucose-6-Phosphate Dehydrogenase Family in Soybean and Functional Identification of GmG6PDH2 Involvement in Salt Stress

et al. 2020

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Glucose-6-phosphate dehydrogenase (G6PDH) is known as a critical enzyme responsible for nicotinamide adenine dinucleotide phosphate (NADPH) generation in the pentose phosphate pathway (PPP), and has an essential function in modulating redox homeostasis and stress responsiveness. In the present work, we characterized the nine members of the G6PDH gene family in soybean. Phylogenic analysis and transit peptide prediction showed that these soybean G6PDHs are divided into plastidic (P) and cytosolic (Cy) isoforms. The subcellular locations of five GmG6PDHs were further verified by confocal microscopy in Arabidopsis mesophyll protoplasts. The respective GmG6PDH genes had distinct expression patterns in various soybean tissues and at different times during seed development. Among them, the Cy-G6PDHs were strongly expressed in roots, developing seeds and nodules, while the transcripts of P-G6PDHs were mainly detected in green tissues. In addition, the activities and transcripts of GmG6PDHs were dramatically stimulated by different stress treatments, including salt, osmotic and alkali. Notably, the expression levels of a cytosolic isoform (GmG6PDH2) were extraordinarily high under salt stress and correlated well with the G6PDH enzyme activities, possibly implying a crucial factor for soybean responses to salinity. Enzymatic assay of recombinant GmG6PDH2 proteins expressed in Escherichia coli showed that the enzyme encoded by GmG6PDH2 had functional NADP +-dependent G6PDH activity. Further analysis indicated overexpression of GmG6PDH2 gene could significantly enhance the resistance of transgenic soybean to salt stress by coordinating with the redox states of ascorbic acid and glutathione pool to suppress reactive oxygen species generation. Together, these results indicate that GmG6PDH2 might be the major isoform for NADPH production in PPP, which is involved in the modulation of cellular AsA-GSH cycle to prevent the oxidative damage induced by high salinity.

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Section: Gmg6pdh2 Is Essential For Asa and Gsh Biosynthesis Under Salmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Genome-Wide Analysis of the Glucose-6-Phosphate Dehydrogenase Family in Soybean and Functional Identification of GmG6PDH2 Involvement in Salt Stress

et al. 2020

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“…Proteins were separated using SDS Gel and then transferred on a nitrocellulose membrane as previously described [17,37]. Filters were incubated with the specific antibodies and reacting polypeptides were revealed by enhanced chemiluminescence using the ECL Prime Kit (Ge Healthcare, Chicago, IL, USA) or (WesternBright Quantum kit-Advansta, San Josè, CA, USA).…”

Section: Proteins Determination and Immunoblottingmentioning

confidence: 99%

Physiological and Molecular Osmotic Stress Responses in Three Durum Wheat (Triticum Turgidum ssp Durum) Genotypes

et al. 2019

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This study aims to investigate the activities and expression of enzymes of primary metabolism and relate these data with the growth performance of three different durum wheat genotypes (Maali; YT13; and ON66) under osmotic stress. Growth traits-including plant height, dry weight (DW) and relative water content (RWC)-were measured to classify genotypes depending on their tolerance to stress. Several enzymes were investigated: Ascorbate peroxidase (APX), Glutamine Synthetase (GS), Glutamine dehydrogenase (GDH), Glutamate synthase (GOGAT), Glucose 6-phosphate dehydrogenase (G6PDH), and Phosphoenolpyruvate Carboxylase (PEPC). The expression of the cytosolic and plastidic glutamine synthetase (TaGS1 and TaGS2), high affinity nitrate transporters (TaNRT2.3) and Glutamate dehydrogenase (TaGDH) were also detected by qRT-PCR. The results indicated different growth performances among genotypes, indicating Maali and YT13 as tolerant genotypes and ON66 as a drought-susceptible variety. Data showed a decrease in PEPC and increase in APX activities under osmotic stress; a slight decrease in GS activity was observed, together with an increase in G6PDH in all genotypes; GS and NRT2 expressions changed in a similar pattern in the different genotypes. Interestingly, Maali and YT13 showed higher transcript abundance for GDH under stress compared to ON66, suggesting the implication of GDH in protective phenomena upon osmotic stress.Agronomy 2019, 9, 550 2 of 15 the photosynthetic activity by its direct effect on CO 2 uptake, resulting in a significant reduction in biomass and shoot growth [2][3][4]. At the cellular level, water scarcity triggers a set of modifications in biological processes caused by modifications in gene expression [5]. An integrated comprehension of morphological and physiological responses to drought requires a deep knowledge of both biochemical and molecular diversity among different genotypes. Indeed, distinct behaviors were observed in susceptible and tolerant genotypes resulting in changes in biomass production, plant height, and greenness [3,6]; these variations are consequences of many visible responses, such as osmolytes syntheses, enzymatic activities, antioxidant machinery, and specific gene expression [3,[7][8][9].In response to water limitation, plants tend to reduce damages by maintaining an optimum water status; to this aim, a set of compatible solutes are synthesized, namely water-soluble carbohydrates (WSC). These components help cells to maintain their turgor. In this context, it has been shown that tolerant genotypes accumulated more WSC than drought-susceptible ones [10].Water scarcity affects different enzymatic activities; among them, the nitrogen metabolism machinery plays a pivotal role in nitrogen utilization [3,8,11]. The activity of the enzymes involved in the GS-GOGAT cycle, a key pathway in nitrogen remobilization, was shown to be widely affected by water stress. Glutamine synthetase (GS), with its two isoforms (cytosolic GS1, and plastidic GS2), is a potential indicator of plant nutri...

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“…G6PDHs from living organisms have been characterized from a number of sources, and kinetic properties of plant plastidic isoforms have also been described previously 12 , 14 , 18 – 21 .…”

Section: Introductionmentioning

confidence: 99%

“… 26 demonstrated that, in chloroplastic P1-G6PDH, the formation of the regulatory disulfide bridge influences Arg 131 position and NADP + correct positioning. The comparison with human G6PDH suggests that this loop “guards” the access to NADP + cofactor molecule to the active site, and possibly would be able to regulate G6PDH activity 15 , 21 . Interestingly, Pt P2-G6PDH mutants in the regulatory cysteines (C175-C183 21 ) lack of redox regulation and show a low activity.…”

Section: Introductionmentioning

confidence: 99%

Mechanism(s) of action of heavy metals to investigate the regulation of plastidic glucose-6-phosphate dehydrogenase

Lillo

Cardi

Landi

et al. 2018

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The regulation of recombinant plastidic glucose-6P dehydrogenase from Populus trichocarpa (PtP2-G6PDH - EC 1.1.1.49) was investigated by exposing wild type and mutagenized isoforms to heavy metals. Nickel and Cadmium caused a marked decrease in PtP2-G6PDH WT activity, suggesting their poisoning effect on plant enzymes; Lead (Pb++) was substantially ineffective. Copper (Cu++) and Zinc (Zn++) exposition resulted in strongest decrease in enzyme activity, thus suggesting a physiological competition with Magnesium, a well-known activator of G6PDH activity. Kinetic analyses confirmed a competitive inhibition by Copper, and a mixed inhibition by (Cd++). Mutagenized enzymes were differently affected by HMs: the reduction of disulfide (C175–C183) exposed the NADP+ binding sites to metals; C145 participates to NADP+ cofactor binding; C194 and C242 are proposed to play a role in the regulation of NADP+/NADPH binding. Copper (and possibly Zinc) is able to occupy competitively Magnesium (Mg++) sites and/or bind to NADP+, resulting in a reduced access of NADP+ sites on the enzyme. Hence, heavy metals could be used to describe specific roles of cysteine residues present in the primary protein sequence; these results are discussed to define the biochemical mechanism(s) of inhibition of plant plastidic G6PDH.

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Plastidic P2 glucose-6P dehydrogenase from poplar is modulated by thioredoxin m-type: Distinct roles of cysteine residues in redox regulation and NADPH inhibition

Cited by 24 publications

References 39 publications

Genome-Wide Analysis of the Glucose-6-Phosphate Dehydrogenase Family in Soybean and Functional Identification of GmG6PDH2 Involvement in Salt Stress

Genome-Wide Analysis of the Glucose-6-Phosphate Dehydrogenase Family in Soybean and Functional Identification of GmG6PDH2 Involvement in Salt Stress

Physiological and Molecular Osmotic Stress Responses in Three Durum Wheat (Triticum Turgidum ssp Durum) Genotypes

Mechanism(s) of action of heavy metals to investigate the regulation of plastidic glucose-6-phosphate dehydrogenase

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