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Abstract:

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.

Registro:

Documento: Artículo
Título:Inhibitory effect of AP-1 complex on 5-aminolevulinate synthase gene expression through sequestration of cAMP-response element protein (CRE)-binding protein (CBP) coactivator
Autor:Guberman, A.S.; Scassa, M.E.; Giono, L.E.; Varone, C.L.; Cánepa, E.T.
Filiación:Departamento de Quimica Biologica, Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, 1428 Buenos Aires, Argentina
Palabras clave: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
Año:2003
Volumen:278
Número:4
Página de inicio:2317
Página de fin:2326
DOI: http://dx.doi.org/10.1074/jbc.M205057200
Título revista:Journal of Biological Chemistry
Título revista abreviado:J. Biol. Chem.
ISSN:00219258
CODEN:JBCHA
CAS:5 aminolevulinate synthase, 9037-14-3; chloramphenicol acetyltransferase, 9040-07-7; cyclic AMP responsive element binding protein, 130428-87-4, 130939-96-7; heme, 14875-96-8; phorbol 13 acetate 12 myristate, 16561-29-8; RNA, 63231-63-0; stress activated protein kinase, 155215-87-5; calphostin C, 121263-19-2; cyclic AMP responsive element binding protein binding protein, 190209-80-4; cyclic AMP, 60-92-4; 5-Aminolevulinate Synthetase, EC 2.3.1.37; calphostin C, 121263-19-2; CREB-Binding Protein, EC 2.3.1.48; CREBBP protein, human; Cyclic AMP, 60-92-4; Naphthalenes; Nuclear Proteins; Proto-Oncogene Proteins c-fos; RNA, Messenger; Trans-Activators; Transcription Factor AP-1
PDF:https://bibliotecadigital.exactas.uba.ar/download/paper/paper_00219258_v278_n4_p2317_Guberman.pdf
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v278_n4_p2317_Guberman

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Citas:

---------- APA ----------
Guberman, A.S., Scassa, M.E., Giono, L.E., Varone, C.L. & Cánepa, E.T. (2003) . Inhibitory effect of AP-1 complex on 5-aminolevulinate synthase gene expression through sequestration of cAMP-response element protein (CRE)-binding protein (CBP) coactivator. Journal of Biological Chemistry, 278(4), 2317-2326.
http://dx.doi.org/10.1074/jbc.M205057200
---------- CHICAGO ----------
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" . Journal of Biological Chemistry 278, no. 4 (2003) : 2317-2326.
http://dx.doi.org/10.1074/jbc.M205057200
---------- MLA ----------
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" . Journal of Biological Chemistry, vol. 278, no. 4, 2003, pp. 2317-2326.
http://dx.doi.org/10.1074/jbc.M205057200
---------- VANCOUVER ----------
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. J. Biol. Chem. 2003;278(4):2317-2326.
http://dx.doi.org/10.1074/jbc.M205057200