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...
| Authors: | ; ; ; ; ; ; ; ; ; |
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| Format: | Article |
| Status: | Published version |
| Publication date: | 2004 |
| Country: | España |
| Institution: | Universidad de Barcelona |
| Repository: | Dipòsit Digital de la UB |
| OAI Identifier: | oai:diposit.ub.edu:2445/181308 |
| Online Access: | https://hdl.handle.net/2445/181308 |
| Access Level: | Open access |
| Keyword: | Genètica Proteïnes Factors de transcripció Genetics Proteins Transcription factors |
| _version_ | 1878738419084951552 |
|---|---|
| author | Obach Cortadellas, Mercè |
| 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 |
| author_browse | Bartrons Bach, Ramon Caro, Jaime Duran i Ferrer, Joan, 1978- Gomez Grau, Marta Kong, Xianguo Navarro i Sabaté, Àurea Obach Cortadellas, Mercè Perales Losa, Carlos Rosa López, José Luis Ventura Pujol, Francesc |
| 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 |
| collection | Dipòsit Digital de la UB |
| 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. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | ES_aead0b145cf2e9e45fcbd3f6f7d2ff2a |
| institution | Universidad de Barcelona |
| instname_str | Universidad de Barcelona |
| network_acronym_str | ES |
| network_name_str | España |
| oai_identifier_str | oai:diposit.ub.edu:2445/181308 |
| publishDate | 2004 |
| publishDateSort | 2004 |
| publisher | American Society for Biochemistry and Molecular Biology |
| publisherStr | American Society for Biochemistry and Molecular Biology |
| reponame_str | Dipòsit Digital de la UB |
| spelling | 6-phosphofructo-2-kinase (pfkfb3) gene promoter contains hypoxia-inducible factor-1 binding sites necessary for transactivation in response to hypoxia 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 Genètica Proteïnes Factors de transcripció Genetics Proteins Transcription factors 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. American Society for Biochemistry and Molecular Biology https://hdl.handle.net/2445/181308 |
| 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 |
| status_str | publishedVersion |
| title | 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_short | 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 |
| topic | Genètica Proteïnes Factors de transcripció Genetics Proteins Transcription factors |
| topic_facet | Genètica Proteïnes Factors de transcripció Genetics Proteins Transcription factors |
| url | https://hdl.handle.net/2445/181308 |