Characterization of the NADP-malic enzymes in the woody plant Populus trichocarpa.
Identifieur interne : 002763 ( Main/Exploration ); précédent : 002762; suivant : 002764Characterization of the NADP-malic enzymes in the woody plant Populus trichocarpa.
Auteurs : Qiguo Yu [République populaire de Chine] ; Jinwen Liu ; Zhifeng Wang ; Jiefei Nai ; Mengyan Lü ; Xiying Zhou ; Yuxiang ChengSource :
- Molecular biology reports [ 1573-4978 ] ; 2013.
Descripteurs français
- KwdFr :
- Analyse de séquence d'ADN (MeSH), Cinétique (MeSH), Concentration en ions d'hydrogène (MeSH), Escherichia coli (MeSH), Expression des gènes (MeSH), Malate dehydrogenase (NADP+) (biosynthèse), Malate dehydrogenase (NADP+) (composition chimique), Malate dehydrogenase (NADP+) (génétique), Malates (composition chimique), Phylogenèse (MeSH), Populus (enzymologie), Populus (génétique), Protéines recombinantes (biosynthèse), Protéines recombinantes (composition chimique), Protéines recombinantes (génétique), Protéines végétales (biosynthèse), Protéines végétales (composition chimique), Protéines végétales (génétique), Régulation de l'expression des gènes au cours du développement (MeSH), Régulation de l'expression des gènes végétaux (MeSH), Similitude de séquences d'acides aminés (MeSH), Stress physiologique (MeSH), Tolérance au sel (MeSH), Évolution moléculaire (MeSH).
- MESH :
- biosynthèse : Malate dehydrogenase (NADP+), Protéines recombinantes, Protéines végétales.
- composition chimique : Malate dehydrogenase (NADP+), Malates, Protéines recombinantes, Protéines végétales.
- enzymologie : Populus.
- génétique : Malate dehydrogenase (NADP+), Populus, Protéines recombinantes, Protéines végétales.
- Analyse de séquence d'ADN, Cinétique, Concentration en ions d'hydrogène, Escherichia coli, Expression des gènes, Phylogenèse, Régulation de l'expression des gènes au cours du développement, Régulation de l'expression des gènes végétaux, Similitude de séquences d'acides aminés, Stress physiologique, Tolérance au sel, Évolution moléculaire.
English descriptors
- KwdEn :
- Escherichia coli (MeSH), Evolution, Molecular (MeSH), Gene Expression (MeSH), Gene Expression Regulation, Developmental (MeSH), Gene Expression Regulation, Plant (MeSH), Hydrogen-Ion Concentration (MeSH), Kinetics (MeSH), Malate Dehydrogenase (NADP+) (biosynthesis), Malate Dehydrogenase (NADP+) (chemistry), Malate Dehydrogenase (NADP+) (genetics), Malates (chemistry), Phylogeny (MeSH), Plant Proteins (biosynthesis), Plant Proteins (chemistry), Plant Proteins (genetics), Populus (enzymology), Populus (genetics), Recombinant Proteins (biosynthesis), Recombinant Proteins (chemistry), Recombinant Proteins (genetics), Salt Tolerance (MeSH), Sequence Analysis, DNA (MeSH), Sequence Homology, Amino Acid (MeSH), Stress, Physiological (MeSH).
- MESH :
- chemical , biosynthesis : Malate Dehydrogenase (NADP+), Plant Proteins, Recombinant Proteins.
- chemical , chemistry : Malate Dehydrogenase (NADP+), Malates, Plant Proteins, Recombinant Proteins.
- chemical , genetics : Malate Dehydrogenase (NADP+), Plant Proteins, Recombinant Proteins.
- enzymology : Populus.
- genetics : Populus.
- Escherichia coli, Evolution, Molecular, Gene Expression, Gene Expression Regulation, Developmental, Gene Expression Regulation, Plant, Hydrogen-Ion Concentration, Kinetics, Phylogeny, Salt Tolerance, Sequence Analysis, DNA, Sequence Homology, Amino Acid, Stress, Physiological.
Abstract
Plant NADP-malic enzyme (NADP-ME, EC 1.1.1.40) participates in a large number of metabolic pathways, but little is known about the NADP-ME family in woody plants or trees. Here, we characterized the tree Populus trichocarpa NADP-ME (PtNADP-ME) family and the properties of the family members. Five NADP-ME genes (PtNADP-ME1-PtNADP-ME5) were found in the genome of Populus. Semi-quantitative RT-PCR analysis show that the transcription levels of PtNADP-ME1 in lignified stems and roots are clearly higher than in other tissues, and PtNADP-ME2, PtNADP-ME3, PtNADP-ME4 and PtNADP-ME5 are broadly expressed in various tissues. PtNADP-ME gene expression was found to respond to salt and osmotic stresses, and NaCl salts upregulated the transcripts of putative plastidic ones (PtNADP-ME4 and PtNADP-ME5) significantly. Further, the NADP-ME activities of Populus seedlings increased at least two-fold under NaCl, mannitol and PEG treatments. Also, the expression of PtNADP-ME2 and PtNADP-ME3 increased during the course of leaf wounding. Each recombinant PtNADP-ME proteins were expressed and purified from Escherichia coli, respectively. Coomassie brilliant blue and NADP-ME activity staining on native polyacrylamide gels showed different oligomeric states of the recombinant PtNADP-MEs in vitro. Noticeably, the cytosolic PtNADP-ME2 aggregates as octamers and hexadecamers while the plastidic PtNADP-ME4 resembles hexamers and octamers. The four PtNADP-ME proteins except for PtNADP-ME1 have high activities on native polyacrylamide gels including different forms for PtNADP-ME2 (octamers and hexadecamers) or for PtNADP-ME4 (hexamers and octamers). High concentrations of NADP substrate decreased the activities of all PtNADP-MEs slightly, while the malate had no effect on them. The kinetic parameters (V (max), K (m), K (cat), and K (cat)/K (m)) of each isoforms were summarized. Our data show the different effects of metabolites (influx into tricarboxylic acid cycle or Calvin cycle) on the activity of the individual PtNADP-ME in vitro. According to phylogenetic analysis, five PtNADP-MEs are clustered into cytosolic dicot, plastidic dicot, and monocot and dicot cytosolic groups in a phylogenetic tree. These results suggest that woody Populus NADP-ME family have diverse properties, and possible roles are discussed.
DOI: 10.1007/s11033-012-2182-y
PubMed: 23096088
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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<term>Gene Expression Regulation, Developmental (MeSH)</term>
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<term>Salt Tolerance</term>
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<term>Concentration en ions d'hydrogène</term>
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<term>Régulation de l'expression des gènes au cours du développement</term>
<term>Régulation de l'expression des gènes végétaux</term>
<term>Similitude de séquences d'acides aminés</term>
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<front><div type="abstract" xml:lang="en">Plant NADP-malic enzyme (NADP-ME, EC 1.1.1.40) participates in a large number of metabolic pathways, but little is known about the NADP-ME family in woody plants or trees. Here, we characterized the tree Populus trichocarpa NADP-ME (PtNADP-ME) family and the properties of the family members. Five NADP-ME genes (PtNADP-ME1-PtNADP-ME5) were found in the genome of Populus. Semi-quantitative RT-PCR analysis show that the transcription levels of PtNADP-ME1 in lignified stems and roots are clearly higher than in other tissues, and PtNADP-ME2, PtNADP-ME3, PtNADP-ME4 and PtNADP-ME5 are broadly expressed in various tissues. PtNADP-ME gene expression was found to respond to salt and osmotic stresses, and NaCl salts upregulated the transcripts of putative plastidic ones (PtNADP-ME4 and PtNADP-ME5) significantly. Further, the NADP-ME activities of Populus seedlings increased at least two-fold under NaCl, mannitol and PEG treatments. Also, the expression of PtNADP-ME2 and PtNADP-ME3 increased during the course of leaf wounding. Each recombinant PtNADP-ME proteins were expressed and purified from Escherichia coli, respectively. Coomassie brilliant blue and NADP-ME activity staining on native polyacrylamide gels showed different oligomeric states of the recombinant PtNADP-MEs in vitro. Noticeably, the cytosolic PtNADP-ME2 aggregates as octamers and hexadecamers while the plastidic PtNADP-ME4 resembles hexamers and octamers. The four PtNADP-ME proteins except for PtNADP-ME1 have high activities on native polyacrylamide gels including different forms for PtNADP-ME2 (octamers and hexadecamers) or for PtNADP-ME4 (hexamers and octamers). High concentrations of NADP substrate decreased the activities of all PtNADP-MEs slightly, while the malate had no effect on them. The kinetic parameters (V (max), K (m), K (cat), and K (cat)/K (m)) of each isoforms were summarized. Our data show the different effects of metabolites (influx into tricarboxylic acid cycle or Calvin cycle) on the activity of the individual PtNADP-ME in vitro. According to phylogenetic analysis, five PtNADP-MEs are clustered into cytosolic dicot, plastidic dicot, and monocot and dicot cytosolic groups in a phylogenetic tree. These results suggest that woody Populus NADP-ME family have diverse properties, and possible roles are discussed.</div>
</front>
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<ArticleTitle>Characterization of the NADP-malic enzymes in the woody plant Populus trichocarpa.</ArticleTitle>
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<Abstract><AbstractText>Plant NADP-malic enzyme (NADP-ME, EC 1.1.1.40) participates in a large number of metabolic pathways, but little is known about the NADP-ME family in woody plants or trees. Here, we characterized the tree Populus trichocarpa NADP-ME (PtNADP-ME) family and the properties of the family members. Five NADP-ME genes (PtNADP-ME1-PtNADP-ME5) were found in the genome of Populus. Semi-quantitative RT-PCR analysis show that the transcription levels of PtNADP-ME1 in lignified stems and roots are clearly higher than in other tissues, and PtNADP-ME2, PtNADP-ME3, PtNADP-ME4 and PtNADP-ME5 are broadly expressed in various tissues. PtNADP-ME gene expression was found to respond to salt and osmotic stresses, and NaCl salts upregulated the transcripts of putative plastidic ones (PtNADP-ME4 and PtNADP-ME5) significantly. Further, the NADP-ME activities of Populus seedlings increased at least two-fold under NaCl, mannitol and PEG treatments. Also, the expression of PtNADP-ME2 and PtNADP-ME3 increased during the course of leaf wounding. Each recombinant PtNADP-ME proteins were expressed and purified from Escherichia coli, respectively. Coomassie brilliant blue and NADP-ME activity staining on native polyacrylamide gels showed different oligomeric states of the recombinant PtNADP-MEs in vitro. Noticeably, the cytosolic PtNADP-ME2 aggregates as octamers and hexadecamers while the plastidic PtNADP-ME4 resembles hexamers and octamers. The four PtNADP-ME proteins except for PtNADP-ME1 have high activities on native polyacrylamide gels including different forms for PtNADP-ME2 (octamers and hexadecamers) or for PtNADP-ME4 (hexamers and octamers). High concentrations of NADP substrate decreased the activities of all PtNADP-MEs slightly, while the malate had no effect on them. The kinetic parameters (V (max), K (m), K (cat), and K (cat)/K (m)) of each isoforms were summarized. Our data show the different effects of metabolites (influx into tricarboxylic acid cycle or Calvin cycle) on the activity of the individual PtNADP-ME in vitro. According to phylogenetic analysis, five PtNADP-MEs are clustered into cytosolic dicot, plastidic dicot, and monocot and dicot cytosolic groups in a phylogenetic tree. These results suggest that woody Populus NADP-ME family have diverse properties, and possible roles are discussed.</AbstractText>
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<ForeName>Xiying</ForeName>
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<affiliations><list><country><li>République populaire de Chine</li>
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<name sortKey="Liu, Jinwen" sort="Liu, Jinwen" uniqKey="Liu J" first="Jinwen" last="Liu">Jinwen Liu</name>
<name sortKey="Lu, Mengyan" sort="Lu, Mengyan" uniqKey="Lu M" first="Mengyan" last="Lü">Mengyan Lü</name>
<name sortKey="Nai, Jiefei" sort="Nai, Jiefei" uniqKey="Nai J" first="Jiefei" last="Nai">Jiefei Nai</name>
<name sortKey="Wang, Zhifeng" sort="Wang, Zhifeng" uniqKey="Wang Z" first="Zhifeng" last="Wang">Zhifeng Wang</name>
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