6-phosphofructo-2-kinase (pfkfb3) gene promoter contains hypoxia-inducible factor-1 binding sites necessary for transactivation in response to hypoxia

The up-regulation of glycolysis to enhance the production of energy under reduced pO(2) is a hallmark of the hypoxic response. A key regulator of glycolytic flux is fructose-2,6-bisphosphate, and its steady state concentration is regulated by the action of different isozymes product of four genes (p...

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Autores: Obach Cortadellas, Mercè, Navarro i Sabaté, Àurea, Caro, Jaime, Kong, Xianguo, Duran i Ferrer, Joan, 1978-, Gomez Grau, Marta, Perales Losa, Carlos, Ventura Pujol, Francesc, Rosa López, José Luis, Bartrons Bach, Ramon
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2004
País:España
Institución:Universidad de Barcelona
Repositorio:Dipòsit Digital de la UB
OAI Identifier:oai:diposit.ub.edu:2445/181308
Acceso en línea:https://hdl.handle.net/2445/181308
Access Level:acceso abierto
Palabra clave:Genètica
Proteïnes
Factors de transcripció
Genetics
Proteins
Transcription factors
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spelling 6-phosphofructo-2-kinase (pfkfb3) gene promoter contains hypoxia-inducible factor-1 binding sites necessary for transactivation in response to hypoxiaObach Cortadellas, MercèNavarro i Sabaté, ÀureaCaro, JaimeKong, XianguoDuran i Ferrer, Joan, 1978-Gomez Grau, MartaPerales Losa, CarlosVentura Pujol, FrancescRosa López, José LuisBartrons Bach, RamonGenèticaProteïnesFactors de transcripcióGeneticsProteinsTranscription factorsThe up-regulation of glycolysis to enhance the production of energy under reduced pO(2) is a hallmark of the hypoxic response. A key regulator of glycolytic flux is fructose-2,6-bisphosphate, and its steady state concentration is regulated by the action of different isozymes product of four genes (pfkfb1-4). pfkfb3 has been found in proliferating cells and tumors, being induced by hypoxia. To understand the organization of cis-acting sequences that are responsible for the oxygen-regulated pfkfb3 gene, we have studied its 5'-flanking region. Extensive analysis of the 5' pfkfb3 promoter sequence revealed the presence of putative consensus binding sites for various transcription factors that could play an important role in pfkfb3 gene regulation. These DNA consensus sequences included estrogen receptor, hypoxia response element (HRE), early growth response, and specific protein 1 putative binding sites. Promoter deletion analysis as well as putative HREs sequences (wild type and mutated) fused to a c-fos minimal promoter unit constructs demonstrate that the sequence located from -1269 to -1297 relative to the start site is required for hypoxia-inducible factor 1 (HIF-1) induction. The effective binding of HIF-1 transcription factor to the HREs at -1279 and -1288 was corroborated by electrophoretic mobility shift assay and biotinylated oligonucleotide pull-down. In addition, HIF-1alpha null mouse embryo fibroblasts transfected with a full-length pfkfb3 promoter-luciferase reporter construct further demonstrated that HIF-1 protein was critically involved for hypoxia transactivation of this gene. Altogether, these results demonstrate that pfkfb3 is a hypoxia-inducible gene that is stimulated through HIF interaction with the consensus HRE site in its promoter region.American Society for Biochemistry and Molecular Biology2004info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://hdl.handle.net/2445/181308Articles publicats en revistes (Patologia i Terapèutica Experimental)reponame:Dipòsit Digital de la UBinstname:Universidad de BarcelonaInglésReproducció del document publicat a: https://doi.org/10.1074/jbc.M406096200Journal of Biological Chemistry, 2004, vol. 279, num. 51, p. 53562-53570https://doi.org/10.1074/jbc.M406096200(c) American Society for Biochemistry and Molecular Biology, 2004info:eu-repo/semantics/openAccessoai:diposit.ub.edu:2445/1813082026-05-27T06:46:51Z
dc.title.none.fl_str_mv 6-phosphofructo-2-kinase (pfkfb3) gene promoter contains hypoxia-inducible factor-1 binding sites necessary for transactivation in response to hypoxia
title 6-phosphofructo-2-kinase (pfkfb3) gene promoter contains hypoxia-inducible factor-1 binding sites necessary for transactivation in response to hypoxia
spellingShingle 6-phosphofructo-2-kinase (pfkfb3) gene promoter contains hypoxia-inducible factor-1 binding sites necessary for transactivation in response to hypoxia
Obach Cortadellas, Mercè
Genètica
Proteïnes
Factors de transcripció
Genetics
Proteins
Transcription factors
title_short 6-phosphofructo-2-kinase (pfkfb3) gene promoter contains hypoxia-inducible factor-1 binding sites necessary for transactivation in response to hypoxia
title_full 6-phosphofructo-2-kinase (pfkfb3) gene promoter contains hypoxia-inducible factor-1 binding sites necessary for transactivation in response to hypoxia
title_fullStr 6-phosphofructo-2-kinase (pfkfb3) gene promoter contains hypoxia-inducible factor-1 binding sites necessary for transactivation in response to hypoxia
title_full_unstemmed 6-phosphofructo-2-kinase (pfkfb3) gene promoter contains hypoxia-inducible factor-1 binding sites necessary for transactivation in response to hypoxia
title_sort 6-phosphofructo-2-kinase (pfkfb3) gene promoter contains hypoxia-inducible factor-1 binding sites necessary for transactivation in response to hypoxia
dc.creator.none.fl_str_mv Obach Cortadellas, Mercè
Navarro i Sabaté, Àurea
Caro, Jaime
Kong, Xianguo
Duran i Ferrer, Joan, 1978-
Gomez Grau, Marta
Perales Losa, Carlos
Ventura Pujol, Francesc
Rosa López, José Luis
Bartrons Bach, Ramon
author Obach Cortadellas, Mercè
author_facet Obach Cortadellas, Mercè
Navarro i Sabaté, Àurea
Caro, Jaime
Kong, Xianguo
Duran i Ferrer, Joan, 1978-
Gomez Grau, Marta
Perales Losa, Carlos
Ventura Pujol, Francesc
Rosa López, José Luis
Bartrons Bach, Ramon
author_role author
author2 Navarro i Sabaté, Àurea
Caro, Jaime
Kong, Xianguo
Duran i Ferrer, Joan, 1978-
Gomez Grau, Marta
Perales Losa, Carlos
Ventura Pujol, Francesc
Rosa López, José Luis
Bartrons Bach, Ramon
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Genètica
Proteïnes
Factors de transcripció
Genetics
Proteins
Transcription factors
topic Genètica
Proteïnes
Factors de transcripció
Genetics
Proteins
Transcription factors
description The up-regulation of glycolysis to enhance the production of energy under reduced pO(2) is a hallmark of the hypoxic response. A key regulator of glycolytic flux is fructose-2,6-bisphosphate, and its steady state concentration is regulated by the action of different isozymes product of four genes (pfkfb1-4). pfkfb3 has been found in proliferating cells and tumors, being induced by hypoxia. To understand the organization of cis-acting sequences that are responsible for the oxygen-regulated pfkfb3 gene, we have studied its 5'-flanking region. Extensive analysis of the 5' pfkfb3 promoter sequence revealed the presence of putative consensus binding sites for various transcription factors that could play an important role in pfkfb3 gene regulation. These DNA consensus sequences included estrogen receptor, hypoxia response element (HRE), early growth response, and specific protein 1 putative binding sites. Promoter deletion analysis as well as putative HREs sequences (wild type and mutated) fused to a c-fos minimal promoter unit constructs demonstrate that the sequence located from -1269 to -1297 relative to the start site is required for hypoxia-inducible factor 1 (HIF-1) induction. The effective binding of HIF-1 transcription factor to the HREs at -1279 and -1288 was corroborated by electrophoretic mobility shift assay and biotinylated oligonucleotide pull-down. In addition, HIF-1alpha null mouse embryo fibroblasts transfected with a full-length pfkfb3 promoter-luciferase reporter construct further demonstrated that HIF-1 protein was critically involved for hypoxia transactivation of this gene. Altogether, these results demonstrate that pfkfb3 is a hypoxia-inducible gene that is stimulated through HIF interaction with the consensus HRE site in its promoter region.
publishDate 2004
dc.date.none.fl_str_mv 2004
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/181308
url https://hdl.handle.net/2445/181308
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a: https://doi.org/10.1074/jbc.M406096200
Journal of Biological Chemistry, 2004, vol. 279, num. 51, p. 53562-53570
https://doi.org/10.1074/jbc.M406096200
dc.rights.none.fl_str_mv (c) American Society for Biochemistry and Molecular Biology, 2004
info:eu-repo/semantics/openAccess
rights_invalid_str_mv (c) American Society for Biochemistry and Molecular Biology, 2004
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
dc.source.none.fl_str_mv Articles publicats en revistes (Patologia i Terapèutica Experimental)
reponame:Dipòsit Digital de la UB
instname:Universidad de Barcelona
instname_str Universidad de Barcelona
reponame_str Dipòsit Digital de la UB
collection Dipòsit Digital de la UB
repository.name.fl_str_mv
repository.mail.fl_str_mv
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