Transcriptional regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase in the adductor muscle of the oyster Crassostrea gigas during prolonged hypoxia.
International audience The response of Crassostrea gigas to prolonged hypoxia was investigated for the first time by analyzing the metabolic branch point formed by pyruvate kinase (PK) and hosphoenolpyruvate carboxykinase (PEPCK). PK and PEPCK cDNAs were cloned and sequenced. The main functional dom...
Published in: | Journal of Experimental Zoology Part A: Ecological Genetics and Physiology |
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Format: | Article in Journal/Newspaper |
Language: | English |
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2007
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Online Access: | https://hal.science/hal-00180077 https://doi.org/10.1002/jez.390 |
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ftmuseumnhn:oai:HAL:hal-00180077v1 2023-07-30T04:03:02+02:00 Transcriptional regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase in the adductor muscle of the oyster Crassostrea gigas during prolonged hypoxia. Le Moullac, Gilles Bacca, Hélène Huvet, Arnaud Moal, Jeanne Pouvreau, Stéphane van Wormhoudt, Alain Biologie des organismes marins et écosystèmes (BOME) Muséum national d'Histoire naturelle (MNHN)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS) Station de Biologie Marine de Concarneau Direction générale déléguée à la Recherche, à l’Expertise, à la Valorisation et à l’Enseignement-Formation (DGD.REVE) Muséum national d'Histoire naturelle (MNHN)-Muséum national d'Histoire naturelle (MNHN) 2007 https://hal.science/hal-00180077 https://doi.org/10.1002/jez.390 en eng HAL CCSD info:eu-repo/semantics/altIdentifier/doi/10.1002/jez.390 info:eu-repo/semantics/altIdentifier/pmid/17486628 hal-00180077 https://hal.science/hal-00180077 doi:10.1002/jez.390 PUBMED: 17486628 Journal of Experimental Zoology Part A Ecological Genetics and Physiology https://hal.science/hal-00180077 Journal of Experimental Zoology Part A Ecological Genetics and Physiology, 2007, 307 (7), pp.371-382. ⟨10.1002/jez.390⟩ oyster Crassostrea gigas PK PEPCK [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology info:eu-repo/semantics/article Journal articles 2007 ftmuseumnhn https://doi.org/10.1002/jez.390 2023-07-08T20:53:53Z International audience The response of Crassostrea gigas to prolonged hypoxia was investigated for the first time by analyzing the metabolic branch point formed by pyruvate kinase (PK) and hosphoenolpyruvate carboxykinase (PEPCK). PK and PEPCK cDNAs were cloned and sequenced. The main functional domains of the PK sequence, such as the binding sites for ADP/ATP and phosphoenolpyruvate (PEP), were identified whereas the PEPCK sequence showed the specific domain to bind PEP in addition to the kinase-1 and kinase-2 motifs to bind guanosine triphosphate (GTP) and Mg(2+), specific for all PEPCKs. A C-terminal extension was detected for the first time in eukaryota PK. Separation of mitochondrial and cytosolic fraction showed that more than 92% of the PEPCK enzyme activity was cytosolic in gills, digestive gland, mantle and muscle. PK and PEPCK mRNAs and enzyme activities have been measured in muscle during prolonged hypoxia for 20 days. Adaptation of PK in hypoxic muscle at transcriptional level occurred lately by decreasing significantly the PK mRNA level at day 20 while PK enzyme activity was inhibited by the high content of alanine. The PEPCK mRNA ratio in hypoxic muscle significantly increased at day 10 simultaneously to the PEPCK enzyme activity. Succinate accumulation observed at day 10 and day 20 confirmed the anaerobic pathway of muscle metabolism in oyster subjected to hypoxia. Regulation of C. gigas PEPCK in muscle occurred at gene transcription level while PK was first regulated at enzyme level with alanine as allosteric inhibitor, and then at molecular level under a fast effect of hypoxia. Article in Journal/Newspaper Crassostrea gigas Muséum National d'Histoire Naturelle (MNHM): HAL Journal of Experimental Zoology Part A: Ecological Genetics and Physiology 307A 7 371 382 |
institution |
Open Polar |
collection |
Muséum National d'Histoire Naturelle (MNHM): HAL |
op_collection_id |
ftmuseumnhn |
language |
English |
topic |
oyster Crassostrea gigas PK PEPCK [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology |
spellingShingle |
oyster Crassostrea gigas PK PEPCK [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Le Moullac, Gilles Bacca, Hélène Huvet, Arnaud Moal, Jeanne Pouvreau, Stéphane van Wormhoudt, Alain Transcriptional regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase in the adductor muscle of the oyster Crassostrea gigas during prolonged hypoxia. |
topic_facet |
oyster Crassostrea gigas PK PEPCK [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology |
description |
International audience The response of Crassostrea gigas to prolonged hypoxia was investigated for the first time by analyzing the metabolic branch point formed by pyruvate kinase (PK) and hosphoenolpyruvate carboxykinase (PEPCK). PK and PEPCK cDNAs were cloned and sequenced. The main functional domains of the PK sequence, such as the binding sites for ADP/ATP and phosphoenolpyruvate (PEP), were identified whereas the PEPCK sequence showed the specific domain to bind PEP in addition to the kinase-1 and kinase-2 motifs to bind guanosine triphosphate (GTP) and Mg(2+), specific for all PEPCKs. A C-terminal extension was detected for the first time in eukaryota PK. Separation of mitochondrial and cytosolic fraction showed that more than 92% of the PEPCK enzyme activity was cytosolic in gills, digestive gland, mantle and muscle. PK and PEPCK mRNAs and enzyme activities have been measured in muscle during prolonged hypoxia for 20 days. Adaptation of PK in hypoxic muscle at transcriptional level occurred lately by decreasing significantly the PK mRNA level at day 20 while PK enzyme activity was inhibited by the high content of alanine. The PEPCK mRNA ratio in hypoxic muscle significantly increased at day 10 simultaneously to the PEPCK enzyme activity. Succinate accumulation observed at day 10 and day 20 confirmed the anaerobic pathway of muscle metabolism in oyster subjected to hypoxia. Regulation of C. gigas PEPCK in muscle occurred at gene transcription level while PK was first regulated at enzyme level with alanine as allosteric inhibitor, and then at molecular level under a fast effect of hypoxia. |
author2 |
Biologie des organismes marins et écosystèmes (BOME) Muséum national d'Histoire naturelle (MNHN)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS) Station de Biologie Marine de Concarneau Direction générale déléguée à la Recherche, à l’Expertise, à la Valorisation et à l’Enseignement-Formation (DGD.REVE) Muséum national d'Histoire naturelle (MNHN)-Muséum national d'Histoire naturelle (MNHN) |
format |
Article in Journal/Newspaper |
author |
Le Moullac, Gilles Bacca, Hélène Huvet, Arnaud Moal, Jeanne Pouvreau, Stéphane van Wormhoudt, Alain |
author_facet |
Le Moullac, Gilles Bacca, Hélène Huvet, Arnaud Moal, Jeanne Pouvreau, Stéphane van Wormhoudt, Alain |
author_sort |
Le Moullac, Gilles |
title |
Transcriptional regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase in the adductor muscle of the oyster Crassostrea gigas during prolonged hypoxia. |
title_short |
Transcriptional regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase in the adductor muscle of the oyster Crassostrea gigas during prolonged hypoxia. |
title_full |
Transcriptional regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase in the adductor muscle of the oyster Crassostrea gigas during prolonged hypoxia. |
title_fullStr |
Transcriptional regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase in the adductor muscle of the oyster Crassostrea gigas during prolonged hypoxia. |
title_full_unstemmed |
Transcriptional regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase in the adductor muscle of the oyster Crassostrea gigas during prolonged hypoxia. |
title_sort |
transcriptional regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase in the adductor muscle of the oyster crassostrea gigas during prolonged hypoxia. |
publisher |
HAL CCSD |
publishDate |
2007 |
url |
https://hal.science/hal-00180077 https://doi.org/10.1002/jez.390 |
genre |
Crassostrea gigas |
genre_facet |
Crassostrea gigas |
op_source |
Journal of Experimental Zoology Part A Ecological Genetics and Physiology https://hal.science/hal-00180077 Journal of Experimental Zoology Part A Ecological Genetics and Physiology, 2007, 307 (7), pp.371-382. ⟨10.1002/jez.390⟩ |
op_relation |
info:eu-repo/semantics/altIdentifier/doi/10.1002/jez.390 info:eu-repo/semantics/altIdentifier/pmid/17486628 hal-00180077 https://hal.science/hal-00180077 doi:10.1002/jez.390 PUBMED: 17486628 |
op_doi |
https://doi.org/10.1002/jez.390 |
container_title |
Journal of Experimental Zoology Part A: Ecological Genetics and Physiology |
container_volume |
307A |
container_issue |
7 |
container_start_page |
371 |
op_container_end_page |
382 |
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1772813963737694208 |