Inhibitory effect of AP-1 complex on 5-aminolevulinate synthase gene expression through sequestration of cAMP-response element protein (CRE)-binding protein (CBP) coactivator
Activation protein-1 (AP-1) transcription factors are early response genes involved in a diverse set of transcriptional regulatory processes. The phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) is often used to induce AP-1 activity. The purpose of this work was to explore the molecular mech...
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| Autores principales: | , , , , |
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| Formato: | Artículo publishedVersion |
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2003
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| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_00219258_v278_n4_p2317_Guberman http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=artiaex&d=paper_00219258_v278_n4_p2317_Guberman_oai |
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I28-R145-paper_00219258_v278_n4_p2317_Guberman_oai |
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| record_format |
dspace |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-145 |
| collection |
Repositorio Digital de la Universidad de Buenos Aires (UBA) |
| topic |
Bioassay Biosynthesis Cloning Esters Proteins Molecular mechanisms Genes 5 aminolevulinate synthase chloramphenicol acetyltransferase cyclic AMP responsive element binding protein heme phorbol 13 acetate 12 myristate RNA stress activated protein kinase transcription factor transcription factor AP 1 calphostin C CREBBP protein, human cyclic AMP cyclic AMP responsive element binding protein binding protein messenger RNA naphthalene derivative nuclear protein protein c fos transactivator protein article biosynthesis controlled study cyclic AMP responsive element DNA flanking region early response gene gene expression genetic transcription hepatoma cell human human cell Northern blotting plasmid priority journal promoter region reporter gene transcription regulation biological model cell culture dimerization dominant gene dose response gene deletion gene vector genetic transfection genetics metabolism molecular cloning protein binding serodiagnosis site directed mutagenesis time Western blotting Butea monosperma Felis catus vectors 5-Aminolevulinate Synthetase Blotting, Western Cloning, Molecular CREB-Binding Protein Cyclic AMP Dimerization Dose-Response Relationship, Drug Gene Deletion Genes, Dominant Genes, Reporter Genetic Vectors Humans Models, Biological Mutagenesis, Site-Directed Naphthalenes Nuclear Proteins Precipitin Tests Promoter Regions (Genetics) Protein Binding Proto-Oncogene Proteins c-fos RNA, Messenger Time Factors Trans-Activators Transcription Factor AP-1 Transcription, Genetic Transfection Tumor Cells, Cultured |
| spellingShingle |
Bioassay Biosynthesis Cloning Esters Proteins Molecular mechanisms Genes 5 aminolevulinate synthase chloramphenicol acetyltransferase cyclic AMP responsive element binding protein heme phorbol 13 acetate 12 myristate RNA stress activated protein kinase transcription factor transcription factor AP 1 calphostin C CREBBP protein, human cyclic AMP cyclic AMP responsive element binding protein binding protein messenger RNA naphthalene derivative nuclear protein protein c fos transactivator protein article biosynthesis controlled study cyclic AMP responsive element DNA flanking region early response gene gene expression genetic transcription hepatoma cell human human cell Northern blotting plasmid priority journal promoter region reporter gene transcription regulation biological model cell culture dimerization dominant gene dose response gene deletion gene vector genetic transfection genetics metabolism molecular cloning protein binding serodiagnosis site directed mutagenesis time Western blotting Butea monosperma Felis catus vectors 5-Aminolevulinate Synthetase Blotting, Western Cloning, Molecular CREB-Binding Protein Cyclic AMP Dimerization Dose-Response Relationship, Drug Gene Deletion Genes, Dominant Genes, Reporter Genetic Vectors Humans Models, Biological Mutagenesis, Site-Directed Naphthalenes Nuclear Proteins Precipitin Tests Promoter Regions (Genetics) Protein Binding Proto-Oncogene Proteins c-fos RNA, Messenger Time Factors Trans-Activators Transcription Factor AP-1 Transcription, Genetic Transfection Tumor Cells, Cultured Guberman, A.S. Scassa, M.E. Giono, L.E. Varone, C.L. Cánepa, E.T. Inhibitory effect of AP-1 complex on 5-aminolevulinate synthase gene expression through sequestration of cAMP-response element protein (CRE)-binding protein (CBP) coactivator |
| topic_facet |
Bioassay Biosynthesis Cloning Esters Proteins Molecular mechanisms Genes 5 aminolevulinate synthase chloramphenicol acetyltransferase cyclic AMP responsive element binding protein heme phorbol 13 acetate 12 myristate RNA stress activated protein kinase transcription factor transcription factor AP 1 calphostin C CREBBP protein, human cyclic AMP cyclic AMP responsive element binding protein binding protein messenger RNA naphthalene derivative nuclear protein protein c fos transactivator protein article biosynthesis controlled study cyclic AMP responsive element DNA flanking region early response gene gene expression genetic transcription hepatoma cell human human cell Northern blotting plasmid priority journal promoter region reporter gene transcription regulation biological model cell culture dimerization dominant gene dose response gene deletion gene vector genetic transfection genetics metabolism molecular cloning protein binding serodiagnosis site directed mutagenesis time Western blotting Butea monosperma Felis catus vectors 5-Aminolevulinate Synthetase Blotting, Western Cloning, Molecular CREB-Binding Protein Cyclic AMP Dimerization Dose-Response Relationship, Drug Gene Deletion Genes, Dominant Genes, Reporter Genetic Vectors Humans Models, Biological Mutagenesis, Site-Directed Naphthalenes Nuclear Proteins Precipitin Tests Promoter Regions (Genetics) Protein Binding Proto-Oncogene Proteins c-fos RNA, Messenger Time Factors Trans-Activators Transcription Factor AP-1 Transcription, Genetic Transfection Tumor Cells, Cultured |
| description |
Activation protein-1 (AP-1) transcription factors are early response genes involved in a diverse set of transcriptional regulatory processes. The phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) is often used to induce AP-1 activity. The purpose of this work was to explore the molecular mechanisms involved in the TPA regulation of ubiquitous 5-aminolevulinate synthase (ALAS) gene expression, the first and rate-controlling step of the heme biosynthesis. Previous analysis of the 5′-flanking sequence of ALAS revealed the existence of two cAMP-response elements (CRE) required for basal and cAMP-stimulated expression. The fragment -833 to +42 in the 5′-flanking region of rat ALAS gene was subcloned into a chloramphenicol acetyltransferase (CAT) reporter vector. The expression vector pALAS/CAT produced a significant CAT activity in transiently transfected HepG2 human hepatoma cells, which was repressed by TPA. Sequence and deletion analysis detected a TPA response element (TRE), located between -261 and -255 (TRE-ALAS), that was critical for TPA regulation. We demonstrated that c-Fos, c-Jun, and JunD are involved in TPA inhibitory effect due to their ability to bind TRE-ALAS, evidenced by supershift analysis and their capacity to repress promoter activity in transfection assays. Repression of ALAS promoter activity by TPA treatment or Fos/Jun overexpression was largely relieved when CRE protein-binding protein or p300 was ectopically expressed. When the TRE site was placed in a different context with respect to CRE sites, it appeared to act as a transcriptional enhancer. We propose that the decrease in ALAS basal activity observed in the presence of TPA may reflect a lower ability of this promoter to assemble the productive pre-initiation complex due to CRE protein-binding protein sequestration. We also suggest that the transcriptional properties of this AP-1 site would depend on a spatial-disposition-dependent manner with respect to the CRE sites and to the transcription initiation site. |
| format |
Artículo Artículo publishedVersion |
| author |
Guberman, A.S. Scassa, M.E. Giono, L.E. Varone, C.L. Cánepa, E.T. |
| author_facet |
Guberman, A.S. Scassa, M.E. Giono, L.E. Varone, C.L. Cánepa, E.T. |
| author_sort |
Guberman, A.S. |
| title |
Inhibitory effect of AP-1 complex on 5-aminolevulinate synthase gene expression through sequestration of cAMP-response element protein (CRE)-binding protein (CBP) coactivator |
| title_short |
Inhibitory effect of AP-1 complex on 5-aminolevulinate synthase gene expression through sequestration of cAMP-response element protein (CRE)-binding protein (CBP) coactivator |
| title_full |
Inhibitory effect of AP-1 complex on 5-aminolevulinate synthase gene expression through sequestration of cAMP-response element protein (CRE)-binding protein (CBP) coactivator |
| title_fullStr |
Inhibitory effect of AP-1 complex on 5-aminolevulinate synthase gene expression through sequestration of cAMP-response element protein (CRE)-binding protein (CBP) coactivator |
| title_full_unstemmed |
Inhibitory effect of AP-1 complex on 5-aminolevulinate synthase gene expression through sequestration of cAMP-response element protein (CRE)-binding protein (CBP) coactivator |
| title_sort |
inhibitory effect of ap-1 complex on 5-aminolevulinate synthase gene expression through sequestration of camp-response element protein (cre)-binding protein (cbp) coactivator |
| publishDate |
2003 |
| url |
http://hdl.handle.net/20.500.12110/paper_00219258_v278_n4_p2317_Guberman http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=artiaex&d=paper_00219258_v278_n4_p2317_Guberman_oai |
| work_keys_str_mv |
AT gubermanas inhibitoryeffectofap1complexon5aminolevulinatesynthasegeneexpressionthroughsequestrationofcampresponseelementproteincrebindingproteincbpcoactivator AT scassame inhibitoryeffectofap1complexon5aminolevulinatesynthasegeneexpressionthroughsequestrationofcampresponseelementproteincrebindingproteincbpcoactivator AT gionole inhibitoryeffectofap1complexon5aminolevulinatesynthasegeneexpressionthroughsequestrationofcampresponseelementproteincrebindingproteincbpcoactivator AT varonecl inhibitoryeffectofap1complexon5aminolevulinatesynthasegeneexpressionthroughsequestrationofcampresponseelementproteincrebindingproteincbpcoactivator AT canepaet inhibitoryeffectofap1complexon5aminolevulinatesynthasegeneexpressionthroughsequestrationofcampresponseelementproteincrebindingproteincbpcoactivator |
| _version_ |
1766026556360097792 |
| spelling |
I28-R145-paper_00219258_v278_n4_p2317_Guberman_oai2020-10-19 Guberman, A.S. Scassa, M.E. Giono, L.E. Varone, C.L. Cánepa, E.T. 2003 Activation protein-1 (AP-1) transcription factors are early response genes involved in a diverse set of transcriptional regulatory processes. The phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) is often used to induce AP-1 activity. The purpose of this work was to explore the molecular mechanisms involved in the TPA regulation of ubiquitous 5-aminolevulinate synthase (ALAS) gene expression, the first and rate-controlling step of the heme biosynthesis. Previous analysis of the 5′-flanking sequence of ALAS revealed the existence of two cAMP-response elements (CRE) required for basal and cAMP-stimulated expression. The fragment -833 to +42 in the 5′-flanking region of rat ALAS gene was subcloned into a chloramphenicol acetyltransferase (CAT) reporter vector. The expression vector pALAS/CAT produced a significant CAT activity in transiently transfected HepG2 human hepatoma cells, which was repressed by TPA. Sequence and deletion analysis detected a TPA response element (TRE), located between -261 and -255 (TRE-ALAS), that was critical for TPA regulation. We demonstrated that c-Fos, c-Jun, and JunD are involved in TPA inhibitory effect due to their ability to bind TRE-ALAS, evidenced by supershift analysis and their capacity to repress promoter activity in transfection assays. Repression of ALAS promoter activity by TPA treatment or Fos/Jun overexpression was largely relieved when CRE protein-binding protein or p300 was ectopically expressed. When the TRE site was placed in a different context with respect to CRE sites, it appeared to act as a transcriptional enhancer. We propose that the decrease in ALAS basal activity observed in the presence of TPA may reflect a lower ability of this promoter to assemble the productive pre-initiation complex due to CRE protein-binding protein sequestration. We also suggest that the transcriptional properties of this AP-1 site would depend on a spatial-disposition-dependent manner with respect to the CRE sites and to the transcription initiation site. Fil:Guberman, A.S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Scassa, M.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Giono, L.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Varone, C.L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Cánepa, E.T. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. application/pdf http://hdl.handle.net/20.500.12110/paper_00219258_v278_n4_p2317_Guberman info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar J. Biol. Chem. 2003;278(4):2317-2326 Bioassay Biosynthesis Cloning Esters Proteins Molecular mechanisms Genes 5 aminolevulinate synthase chloramphenicol acetyltransferase cyclic AMP responsive element binding protein heme phorbol 13 acetate 12 myristate RNA stress activated protein kinase transcription factor transcription factor AP 1 calphostin C CREBBP protein, human cyclic AMP cyclic AMP responsive element binding protein binding protein messenger RNA naphthalene derivative nuclear protein protein c fos transactivator protein article biosynthesis controlled study cyclic AMP responsive element DNA flanking region early response gene gene expression genetic transcription hepatoma cell human human cell Northern blotting plasmid priority journal promoter region reporter gene transcription regulation biological model cell culture dimerization dominant gene dose response gene deletion gene vector genetic transfection genetics metabolism molecular cloning protein binding serodiagnosis site directed mutagenesis time Western blotting Butea monosperma Felis catus vectors 5-Aminolevulinate Synthetase Blotting, Western Cloning, Molecular CREB-Binding Protein Cyclic AMP Dimerization Dose-Response Relationship, Drug Gene Deletion Genes, Dominant Genes, Reporter Genetic Vectors Humans Models, Biological Mutagenesis, Site-Directed Naphthalenes Nuclear Proteins Precipitin Tests Promoter Regions (Genetics) Protein Binding Proto-Oncogene Proteins c-fos RNA, Messenger Time Factors Trans-Activators Transcription Factor AP-1 Transcription, Genetic Transfection Tumor Cells, Cultured Inhibitory effect of AP-1 complex on 5-aminolevulinate synthase gene expression through sequestration of cAMP-response element protein (CRE)-binding protein (CBP) coactivator info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=artiaex&d=paper_00219258_v278_n4_p2317_Guberman_oai |