Transcriptional regulation of the rat bradykinin B2 receptor gene: Identification of a silencer element
Autor(a) principal: | |
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Data de Publicação: | 2002 |
Outros Autores: | , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/handle/11600/27047 http://dx.doi.org/10.1124/mol.62.6.1344 |
Resumo: | Kinins are involved in a variety of physiological and pathophysiological processes related to cardiovascular homeostasis, inflammation, blood flow, and nociception. Under physiological conditions, the bradykinin B2 (BKB2) receptor is constitutively expressed and mediates most of kinins' actions. However, the mechanisms regulating BKB2 receptor gene expression are still poorly understood. in this study, 4.6 kilobases of the 5'-flanking region from the rat BKB2 receptor gene were sequenced, and computer analysis revealed several sites for transcriptional factors. Nine promoter mutants were cloned in luciferase reporter gene vectors and transfected in NG108-15 cells and rat aorta vascular smooth muscle cells (VSMCs), showing several positive and negative regulatory elements. A classical silencer with 56 base pairs (bp) caused a decrease in reporter gene activity in NG108-15 cells and VSMCs and was able to inhibit the thymidine kinase promoter. Using electrophoretic mobility shift assay and surface plasmon resonance assay, protein-DNA interactions in the silencer region were determined and specific sets of protein-silencer complexes were detected in both cell types. More intense complexes were observed in the central 21 bp of the silencer and mutation in a putative SRE-1 site strongly impaired the protein-DNA binding. Down-regulation of the BKB2 receptor population in NG108-15 cells promoted by N-6, 2'-O-dibutyryladenosine 3':5'-cyclic monophosphate was paralleled by an increase in the amount of nuclear proteins bound to the silencer sequence showing an inverse relationship between protein-silencer complexes and the transcription of the BKB2 receptor gene. in summary, these data highlight the cell-specific regulation of the BKB2 receptor and the importance of a silencer element present in the regulatory region of the gene. |
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Baptista, Heloisa A.Avellar, Maria CWAraujo, Ronaldo C.Pesquero, Jorge L.Schanstra, Joost P.Bascands, Jean L.Esteve, Jean P.Paiva, Antonio CMBader, MichaelPesquero, João Bosco [UNIFESP]Universidade Federal de São Paulo (UNIFESP)Univ Mogi CruzesUniversidade Federal de Minas Gerais (UFMG)Inst Louis BugnardMax Delbruck Ctr Mol Med2016-01-24T12:33:36Z2016-01-24T12:33:36Z2002-12-01Molecular Pharmacology. Bethesda: Amer Soc Pharmacology Experimental Therapeutics, v. 62, n. 6, p. 1344-1355, 2002.0026-895Xhttp://repositorio.unifesp.br/handle/11600/27047http://dx.doi.org/10.1124/mol.62.6.134410.1124/mol.62.6.1344WOS:000179268900010Kinins are involved in a variety of physiological and pathophysiological processes related to cardiovascular homeostasis, inflammation, blood flow, and nociception. Under physiological conditions, the bradykinin B2 (BKB2) receptor is constitutively expressed and mediates most of kinins' actions. However, the mechanisms regulating BKB2 receptor gene expression are still poorly understood. in this study, 4.6 kilobases of the 5'-flanking region from the rat BKB2 receptor gene were sequenced, and computer analysis revealed several sites for transcriptional factors. Nine promoter mutants were cloned in luciferase reporter gene vectors and transfected in NG108-15 cells and rat aorta vascular smooth muscle cells (VSMCs), showing several positive and negative regulatory elements. A classical silencer with 56 base pairs (bp) caused a decrease in reporter gene activity in NG108-15 cells and VSMCs and was able to inhibit the thymidine kinase promoter. Using electrophoretic mobility shift assay and surface plasmon resonance assay, protein-DNA interactions in the silencer region were determined and specific sets of protein-silencer complexes were detected in both cell types. More intense complexes were observed in the central 21 bp of the silencer and mutation in a putative SRE-1 site strongly impaired the protein-DNA binding. Down-regulation of the BKB2 receptor population in NG108-15 cells promoted by N-6, 2'-O-dibutyryladenosine 3':5'-cyclic monophosphate was paralleled by an increase in the amount of nuclear proteins bound to the silencer sequence showing an inverse relationship between protein-silencer complexes and the transcription of the BKB2 receptor gene. in summary, these data highlight the cell-specific regulation of the BKB2 receptor and the importance of a silencer element present in the regulatory region of the gene.Universidade Federal de São Paulo, Dept Biophys, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Pharmacol, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Ctr Dev Expt Models Med & Biol, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Interdisciplinary Ctr Biochem Res, BR-04023062 São Paulo, BrazilUniv Mogi Cruzes, Mogi Das Cruzes, BrazilUniv Fed Minas Gerais, Dept Physiol & Biophys, Belo Horizonte, MG, BrazilInst Louis Bugnard, INSERM, U388, Toulouse, FranceInst Louis Bugnard, INSERM, U531, Toulouse, FranceMax Delbruck Ctr Mol Med, Berlin, GermanyUniversidade Federal de São Paulo, Dept Biophys, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Pharmacol, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Ctr Dev Expt Models Med & Biol, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Interdisciplinary Ctr Biochem Res, BR-04023062 São Paulo, BrazilWeb of Science1344-1355engAmer Soc Pharmacology Experimental TherapeuticsMolecular PharmacologyTranscriptional regulation of the rat bradykinin B2 receptor gene: Identification of a silencer elementinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP11600/270472023-01-30 22:19:12.086metadata only accessoai:repositorio.unifesp.br:11600/27047Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652023-01-31T01:19:12Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Transcriptional regulation of the rat bradykinin B2 receptor gene: Identification of a silencer element |
title |
Transcriptional regulation of the rat bradykinin B2 receptor gene: Identification of a silencer element |
spellingShingle |
Transcriptional regulation of the rat bradykinin B2 receptor gene: Identification of a silencer element Baptista, Heloisa A. |
title_short |
Transcriptional regulation of the rat bradykinin B2 receptor gene: Identification of a silencer element |
title_full |
Transcriptional regulation of the rat bradykinin B2 receptor gene: Identification of a silencer element |
title_fullStr |
Transcriptional regulation of the rat bradykinin B2 receptor gene: Identification of a silencer element |
title_full_unstemmed |
Transcriptional regulation of the rat bradykinin B2 receptor gene: Identification of a silencer element |
title_sort |
Transcriptional regulation of the rat bradykinin B2 receptor gene: Identification of a silencer element |
author |
Baptista, Heloisa A. |
author_facet |
Baptista, Heloisa A. Avellar, Maria CW Araujo, Ronaldo C. Pesquero, Jorge L. Schanstra, Joost P. Bascands, Jean L. Esteve, Jean P. Paiva, Antonio CM Bader, Michael Pesquero, João Bosco [UNIFESP] |
author_role |
author |
author2 |
Avellar, Maria CW Araujo, Ronaldo C. Pesquero, Jorge L. Schanstra, Joost P. Bascands, Jean L. Esteve, Jean P. Paiva, Antonio CM Bader, Michael Pesquero, João Bosco [UNIFESP] |
author2_role |
author author author author author author author author author |
dc.contributor.institution.none.fl_str_mv |
Universidade Federal de São Paulo (UNIFESP) Univ Mogi Cruzes Universidade Federal de Minas Gerais (UFMG) Inst Louis Bugnard Max Delbruck Ctr Mol Med |
dc.contributor.author.fl_str_mv |
Baptista, Heloisa A. Avellar, Maria CW Araujo, Ronaldo C. Pesquero, Jorge L. Schanstra, Joost P. Bascands, Jean L. Esteve, Jean P. Paiva, Antonio CM Bader, Michael Pesquero, João Bosco [UNIFESP] |
description |
Kinins are involved in a variety of physiological and pathophysiological processes related to cardiovascular homeostasis, inflammation, blood flow, and nociception. Under physiological conditions, the bradykinin B2 (BKB2) receptor is constitutively expressed and mediates most of kinins' actions. However, the mechanisms regulating BKB2 receptor gene expression are still poorly understood. in this study, 4.6 kilobases of the 5'-flanking region from the rat BKB2 receptor gene were sequenced, and computer analysis revealed several sites for transcriptional factors. Nine promoter mutants were cloned in luciferase reporter gene vectors and transfected in NG108-15 cells and rat aorta vascular smooth muscle cells (VSMCs), showing several positive and negative regulatory elements. A classical silencer with 56 base pairs (bp) caused a decrease in reporter gene activity in NG108-15 cells and VSMCs and was able to inhibit the thymidine kinase promoter. Using electrophoretic mobility shift assay and surface plasmon resonance assay, protein-DNA interactions in the silencer region were determined and specific sets of protein-silencer complexes were detected in both cell types. More intense complexes were observed in the central 21 bp of the silencer and mutation in a putative SRE-1 site strongly impaired the protein-DNA binding. Down-regulation of the BKB2 receptor population in NG108-15 cells promoted by N-6, 2'-O-dibutyryladenosine 3':5'-cyclic monophosphate was paralleled by an increase in the amount of nuclear proteins bound to the silencer sequence showing an inverse relationship between protein-silencer complexes and the transcription of the BKB2 receptor gene. in summary, these data highlight the cell-specific regulation of the BKB2 receptor and the importance of a silencer element present in the regulatory region of the gene. |
publishDate |
2002 |
dc.date.issued.fl_str_mv |
2002-12-01 |
dc.date.accessioned.fl_str_mv |
2016-01-24T12:33:36Z |
dc.date.available.fl_str_mv |
2016-01-24T12:33:36Z |
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.citation.fl_str_mv |
Molecular Pharmacology. Bethesda: Amer Soc Pharmacology Experimental Therapeutics, v. 62, n. 6, p. 1344-1355, 2002. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/27047 http://dx.doi.org/10.1124/mol.62.6.1344 |
dc.identifier.issn.none.fl_str_mv |
0026-895X |
dc.identifier.doi.none.fl_str_mv |
10.1124/mol.62.6.1344 |
dc.identifier.wos.none.fl_str_mv |
WOS:000179268900010 |
identifier_str_mv |
Molecular Pharmacology. Bethesda: Amer Soc Pharmacology Experimental Therapeutics, v. 62, n. 6, p. 1344-1355, 2002. 0026-895X 10.1124/mol.62.6.1344 WOS:000179268900010 |
url |
http://repositorio.unifesp.br/handle/11600/27047 http://dx.doi.org/10.1124/mol.62.6.1344 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.none.fl_str_mv |
Molecular Pharmacology |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
1344-1355 |
dc.publisher.none.fl_str_mv |
Amer Soc Pharmacology Experimental Therapeutics |
publisher.none.fl_str_mv |
Amer Soc Pharmacology Experimental Therapeutics |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UNIFESP instname:Universidade Federal de São Paulo (UNIFESP) instacron:UNIFESP |
instname_str |
Universidade Federal de São Paulo (UNIFESP) |
instacron_str |
UNIFESP |
institution |
UNIFESP |
reponame_str |
Repositório Institucional da UNIFESP |
collection |
Repositório Institucional da UNIFESP |
repository.name.fl_str_mv |
Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP) |
repository.mail.fl_str_mv |
|
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1802764280179720192 |