During the initiation of fermentation overexpression of hexokinase PII in yeast transiently causes a similar deregulation of glycolysis as deletion of Tps1
Autor(a) principal: | |
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Data de Publicação: | 1998 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1002/(SICI)1097-0061(199802)14:3<255 http://hdl.handle.net/11449/65391 |
Resumo: | In the yeast Saccharomyces cerevisiae a novel control exerted by TPS1 (=GGS1=FDP1=BYP1=CIF1=GLC6=TSS1)-encoded trehalose-6-phosphate synthase, is essential for restriction of glucose influx into glycolysis apparently by inhibiting hexokinase activity in vivo. We show that up to 50-fold overexpression of hexokinase does not noticeably affect growth on glucose or fructose in wild-type cells. However, it causes higher levels of glucose-6-phosphate, fructose-6-phosphate and also faster accumulation of fructose-1,6-bisphosphate during the initiation of fermentation. The levels of ATP and Pi correlated inversely with the higher sugar phosphate levels. In the first minutes after glucose addition, the metabolite pattern observed was intermediate between those of the tps1Δ mutant and tile wild-type strain. Apparently, during the start-up of fermentation hexokinase is more rate-limiting in the first section of glycolysis than phosphofructokinase. We have developed a method to measure the free intracellular glucose level which is based on the simultaneous addition of D-glucose and an equal concentration of radiolabelled L-glucose. Since the latter is not transported, the free intracellular glucose level can be calculated as the difference between the total B-glucose measured (intracellular + periplasmic/extracellular) and the total L-glucose measured (periplasmic/extracellular). The intracellular glucose level rose in 5 min after addition of 100 mM-glucose to 0.5-2 mM in the wild-type strain, ± 10 mm in a hxk1Δ hxk2Δ glk1Δ and 2-3 mM in a tps1Δ strain. In the strains overexpressing hexokinase PII the level of free intracellular glucose was not reduced. Overexpression of hexokinase PII never produced a strong effect on the rate of ethanol production and glucose consumption. Our results show that overexpression of hexokinase does not cause the same phenotype as deletion of Tps1. However, it mimics it transiently during the initiation of fermentation. Afterwards, the Tps1-dependent control system is apparently able to restrict Properly up to 50-fold higher hexokinase activity. |
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During the initiation of fermentation overexpression of hexokinase PII in yeast transiently causes a similar deregulation of glycolysis as deletion of Tps1FermentationGlycolysisHexokinase PIITps1hexokinasecontrol systemcontrolled studyderegulationenzyme activityfermentationgene overexpressionglucose metabolismglycolysisnonhumanpriority journalsaccharomyces cerevisiaeAdenosine TriphosphateEthanolFungal ProteinsGene DeletionGenes, FungalGlucoseHexokinasePhosphotransferasesPolymerase Chain ReactionSaccharomyces cerevisiaeSpecies SpecificityIn the yeast Saccharomyces cerevisiae a novel control exerted by TPS1 (=GGS1=FDP1=BYP1=CIF1=GLC6=TSS1)-encoded trehalose-6-phosphate synthase, is essential for restriction of glucose influx into glycolysis apparently by inhibiting hexokinase activity in vivo. We show that up to 50-fold overexpression of hexokinase does not noticeably affect growth on glucose or fructose in wild-type cells. However, it causes higher levels of glucose-6-phosphate, fructose-6-phosphate and also faster accumulation of fructose-1,6-bisphosphate during the initiation of fermentation. The levels of ATP and Pi correlated inversely with the higher sugar phosphate levels. In the first minutes after glucose addition, the metabolite pattern observed was intermediate between those of the tps1Δ mutant and tile wild-type strain. Apparently, during the start-up of fermentation hexokinase is more rate-limiting in the first section of glycolysis than phosphofructokinase. We have developed a method to measure the free intracellular glucose level which is based on the simultaneous addition of D-glucose and an equal concentration of radiolabelled L-glucose. Since the latter is not transported, the free intracellular glucose level can be calculated as the difference between the total B-glucose measured (intracellular + periplasmic/extracellular) and the total L-glucose measured (periplasmic/extracellular). The intracellular glucose level rose in 5 min after addition of 100 mM-glucose to 0.5-2 mM in the wild-type strain, ± 10 mm in a hxk1Δ hxk2Δ glk1Δ and 2-3 mM in a tps1Δ strain. In the strains overexpressing hexokinase PII the level of free intracellular glucose was not reduced. Overexpression of hexokinase PII never produced a strong effect on the rate of ethanol production and glucose consumption. Our results show that overexpression of hexokinase does not cause the same phenotype as deletion of Tps1. However, it mimics it transiently during the initiation of fermentation. Afterwards, the Tps1-dependent control system is apparently able to restrict Properly up to 50-fold higher hexokinase activity.Lab. voor Moleculaire Celbiologie Katholieke Universiteit Leuven, Kardinaal Mercierlaan 92, B-3001 Leuven-Heverlee, FlandersDepartment of Biochemistry Institute of Chemistry São Paulo State University, C.P. 355, 14800-900 Araraquara, SPLab. Bioquim. Fisiol. Microrganismos Escola de Farmácia Universidade Federal de Ouro Preto, Rua Costa Sena 171, 35.400.00 Ouro Preto, MGDepartment of Biochemistry Institute of Chemistry São Paulo State University, C.P. 355, 14800-900 Araraquara, SPKatholieke Universiteit LeuvenUniversidade Estadual Paulista (Unesp)Universidade Federal de Ouro Preto (UFOP)Ernandes, Jose Roberto [UNESP]De Meirsman, CatherineRolland, FilipWinderickx, JorisDe Winde, JohannesBrandão, Rogelio LopesThevelein, Johan M.2014-05-27T11:19:33Z2014-05-27T11:19:33Z1998-02-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article255-269application/pdfhttp://dx.doi.org/10.1002/(SICI)1097-0061(199802)14:3<255Yeast, v. 14, n. 3, p. 255-269, 1998.0749-503Xhttp://hdl.handle.net/11449/6539110.1002/(SICI)1097-0061(199802)14:3<2552-s2.0-00318917072-s2.0-0031891707.pdf4966823021866296Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengYeast2.283info:eu-repo/semantics/openAccess2023-10-28T06:06:11Zoai:repositorio.unesp.br:11449/65391Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462023-10-28T06:06:11Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
During the initiation of fermentation overexpression of hexokinase PII in yeast transiently causes a similar deregulation of glycolysis as deletion of Tps1 |
title |
During the initiation of fermentation overexpression of hexokinase PII in yeast transiently causes a similar deregulation of glycolysis as deletion of Tps1 |
spellingShingle |
During the initiation of fermentation overexpression of hexokinase PII in yeast transiently causes a similar deregulation of glycolysis as deletion of Tps1 Ernandes, Jose Roberto [UNESP] Fermentation Glycolysis Hexokinase PII Tps1 hexokinase control system controlled study deregulation enzyme activity fermentation gene overexpression glucose metabolism glycolysis nonhuman priority journal saccharomyces cerevisiae Adenosine Triphosphate Ethanol Fungal Proteins Gene Deletion Genes, Fungal Glucose Hexokinase Phosphotransferases Polymerase Chain Reaction Saccharomyces cerevisiae Species Specificity |
title_short |
During the initiation of fermentation overexpression of hexokinase PII in yeast transiently causes a similar deregulation of glycolysis as deletion of Tps1 |
title_full |
During the initiation of fermentation overexpression of hexokinase PII in yeast transiently causes a similar deregulation of glycolysis as deletion of Tps1 |
title_fullStr |
During the initiation of fermentation overexpression of hexokinase PII in yeast transiently causes a similar deregulation of glycolysis as deletion of Tps1 |
title_full_unstemmed |
During the initiation of fermentation overexpression of hexokinase PII in yeast transiently causes a similar deregulation of glycolysis as deletion of Tps1 |
title_sort |
During the initiation of fermentation overexpression of hexokinase PII in yeast transiently causes a similar deregulation of glycolysis as deletion of Tps1 |
author |
Ernandes, Jose Roberto [UNESP] |
author_facet |
Ernandes, Jose Roberto [UNESP] De Meirsman, Catherine Rolland, Filip Winderickx, Joris De Winde, Johannes Brandão, Rogelio Lopes Thevelein, Johan M. |
author_role |
author |
author2 |
De Meirsman, Catherine Rolland, Filip Winderickx, Joris De Winde, Johannes Brandão, Rogelio Lopes Thevelein, Johan M. |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
Katholieke Universiteit Leuven Universidade Estadual Paulista (Unesp) Universidade Federal de Ouro Preto (UFOP) |
dc.contributor.author.fl_str_mv |
Ernandes, Jose Roberto [UNESP] De Meirsman, Catherine Rolland, Filip Winderickx, Joris De Winde, Johannes Brandão, Rogelio Lopes Thevelein, Johan M. |
dc.subject.por.fl_str_mv |
Fermentation Glycolysis Hexokinase PII Tps1 hexokinase control system controlled study deregulation enzyme activity fermentation gene overexpression glucose metabolism glycolysis nonhuman priority journal saccharomyces cerevisiae Adenosine Triphosphate Ethanol Fungal Proteins Gene Deletion Genes, Fungal Glucose Hexokinase Phosphotransferases Polymerase Chain Reaction Saccharomyces cerevisiae Species Specificity |
topic |
Fermentation Glycolysis Hexokinase PII Tps1 hexokinase control system controlled study deregulation enzyme activity fermentation gene overexpression glucose metabolism glycolysis nonhuman priority journal saccharomyces cerevisiae Adenosine Triphosphate Ethanol Fungal Proteins Gene Deletion Genes, Fungal Glucose Hexokinase Phosphotransferases Polymerase Chain Reaction Saccharomyces cerevisiae Species Specificity |
description |
In the yeast Saccharomyces cerevisiae a novel control exerted by TPS1 (=GGS1=FDP1=BYP1=CIF1=GLC6=TSS1)-encoded trehalose-6-phosphate synthase, is essential for restriction of glucose influx into glycolysis apparently by inhibiting hexokinase activity in vivo. We show that up to 50-fold overexpression of hexokinase does not noticeably affect growth on glucose or fructose in wild-type cells. However, it causes higher levels of glucose-6-phosphate, fructose-6-phosphate and also faster accumulation of fructose-1,6-bisphosphate during the initiation of fermentation. The levels of ATP and Pi correlated inversely with the higher sugar phosphate levels. In the first minutes after glucose addition, the metabolite pattern observed was intermediate between those of the tps1Δ mutant and tile wild-type strain. Apparently, during the start-up of fermentation hexokinase is more rate-limiting in the first section of glycolysis than phosphofructokinase. We have developed a method to measure the free intracellular glucose level which is based on the simultaneous addition of D-glucose and an equal concentration of radiolabelled L-glucose. Since the latter is not transported, the free intracellular glucose level can be calculated as the difference between the total B-glucose measured (intracellular + periplasmic/extracellular) and the total L-glucose measured (periplasmic/extracellular). The intracellular glucose level rose in 5 min after addition of 100 mM-glucose to 0.5-2 mM in the wild-type strain, ± 10 mm in a hxk1Δ hxk2Δ glk1Δ and 2-3 mM in a tps1Δ strain. In the strains overexpressing hexokinase PII the level of free intracellular glucose was not reduced. Overexpression of hexokinase PII never produced a strong effect on the rate of ethanol production and glucose consumption. Our results show that overexpression of hexokinase does not cause the same phenotype as deletion of Tps1. However, it mimics it transiently during the initiation of fermentation. Afterwards, the Tps1-dependent control system is apparently able to restrict Properly up to 50-fold higher hexokinase activity. |
publishDate |
1998 |
dc.date.none.fl_str_mv |
1998-02-01 2014-05-27T11:19:33Z 2014-05-27T11:19:33Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1002/(SICI)1097-0061(199802)14:3<255 Yeast, v. 14, n. 3, p. 255-269, 1998. 0749-503X http://hdl.handle.net/11449/65391 10.1002/(SICI)1097-0061(199802)14:3<255 2-s2.0-0031891707 2-s2.0-0031891707.pdf 4966823021866296 |
url |
http://dx.doi.org/10.1002/(SICI)1097-0061(199802)14:3<255 http://hdl.handle.net/11449/65391 |
identifier_str_mv |
Yeast, v. 14, n. 3, p. 255-269, 1998. 0749-503X 10.1002/(SICI)1097-0061(199802)14:3<255 2-s2.0-0031891707 2-s2.0-0031891707.pdf 4966823021866296 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Yeast 2.283 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
255-269 application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
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1803046196626849792 |