Transcriptional regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase in the adductor muscle of the oyster Crassostrea gigas during prolonged hypoxia
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 phosphoenolpyruvate carboxykinase (PEPCK). PK and PEPCK cDNAs were cloned and sequenced. The main functional domains of the PK sequenc...
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ftarchimer:oai:archimer.ifremer.fr:2683 2023-05-15T15:58:22+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, Helene Huvet, Arnaud Moal, Jeanne Pouvreau, Stephane Van Wormhoudt, A 2007-06 application/pdf https://archimer.ifremer.fr/doc/2007/publication-2683.pdf https://doi.org/10.1002/jez.390 https://archimer.ifremer.fr/doc/00000/2683/ eng eng Wiley InterScience https://archimer.ifremer.fr/doc/2007/publication-2683.pdf doi:10.1002/jez.390 https://archimer.ifremer.fr/doc/00000/2683/ 2007 Wiley InterScience info:eu-repo/semantics/openAccess restricted use Journal of Experimental Zoology Part A: Ecological Genetics and Physiology (1932-5223) (Wiley InterScience), 2007-06 , Vol. 307A , N. 7 , P. 371-382 succinate alanine carboxykinase phosphoenolpyruvate pyruvate kinase hypoxia Oyster text Publication info:eu-repo/semantics/article 2007 ftarchimer https://doi.org/10.1002/jez.390 2021-09-23T20:14:53Z 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 phosphoenolpyruvate 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 Mg2+, 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 Archimer (Archive Institutionnelle de l'Ifremer - Institut français de recherche pour l'exploitation de la mer) Journal of Experimental Zoology Part A: Ecological Genetics and Physiology 307A 7 371 382 |
institution |
Open Polar |
collection |
Archimer (Archive Institutionnelle de l'Ifremer - Institut français de recherche pour l'exploitation de la mer) |
op_collection_id |
ftarchimer |
language |
English |
topic |
succinate alanine carboxykinase phosphoenolpyruvate pyruvate kinase hypoxia Oyster |
spellingShingle |
succinate alanine carboxykinase phosphoenolpyruvate pyruvate kinase hypoxia Oyster Le Moullac, Gilles Bacca, Helene Huvet, Arnaud Moal, Jeanne Pouvreau, Stephane Van Wormhoudt, A Transcriptional regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase in the adductor muscle of the oyster Crassostrea gigas during prolonged hypoxia |
topic_facet |
succinate alanine carboxykinase phosphoenolpyruvate pyruvate kinase hypoxia Oyster |
description |
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 phosphoenolpyruvate 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 Mg2+, 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. |
format |
Article in Journal/Newspaper |
author |
Le Moullac, Gilles Bacca, Helene Huvet, Arnaud Moal, Jeanne Pouvreau, Stephane Van Wormhoudt, A |
author_facet |
Le Moullac, Gilles Bacca, Helene Huvet, Arnaud Moal, Jeanne Pouvreau, Stephane Van Wormhoudt, A |
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 |
Wiley InterScience |
publishDate |
2007 |
url |
https://archimer.ifremer.fr/doc/2007/publication-2683.pdf https://doi.org/10.1002/jez.390 https://archimer.ifremer.fr/doc/00000/2683/ |
genre |
Crassostrea gigas |
genre_facet |
Crassostrea gigas |
op_source |
Journal of Experimental Zoology Part A: Ecological Genetics and Physiology (1932-5223) (Wiley InterScience), 2007-06 , Vol. 307A , N. 7 , P. 371-382 |
op_relation |
https://archimer.ifremer.fr/doc/2007/publication-2683.pdf doi:10.1002/jez.390 https://archimer.ifremer.fr/doc/00000/2683/ |
op_rights |
2007 Wiley InterScience info:eu-repo/semantics/openAccess restricted use |
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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1766394106855030784 |