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Article: Sp1 and Sp3 transcription factors are required for trans-activation of the human SERCA2 promoter in cardiomyocytes

TitleSp1 and Sp3 transcription factors are required for trans-activation of the human SERCA2 promoter in cardiomyocytes
Authors
Issue Date2003
Citation
Cardiovascular Research, 2003, v. 60 n. 2, p. 347-354 How to Cite?
AbstractObjectives: The sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA) is essential to the removal of cytosolic calcium following cardiac contraction, and its abundance and activity are significantly altered during perinatal development and in failing myocardium. The objective of the current study was to identify cis regulatory elements and nuclear transcription factors responsible for transactivating SERCA2 gene expression in cardiomyocytes. Methods: Primary cultures of neonatal rat ventricular myocytes were transiently transfected with luciferase (LUX) reporter gene constructs containing deletions of the SERCA2 promoter or which harbored mutations in consensus Sp1 transcription factor binding sites. Cotransfection assays, electrophoretic mobility shift, and supershift assays were also performed to delineate the regulatory role of specific transcription factors. Results: We identified a putative AP-1-like element and a consensus Egr-1 binding site, but neither Egr-1 nor 12-O-tetradecanoylphorbol 13-acetate (TPA) significantly modified human SERCA2 promoter activity in vitro. Maximal activity of the SERCA2 promoter required the proximal 177 bp, and strong activation was observed with a 125-bp construct, within which an Sp1 site and a CAAT box were important. Mutation analysis also revealed the importance of two Sp1 sites between -125 and -200. Sp1 and Sp3 transcription factors were subsequently identified to bind to oligonucleotide probes corresponding to only the two most proximal Sp1 sites. Conclusions: These studies provide direct evidence that regulation of human SERCA2 gene expression in cardiomyocytes depends on transactivation events elicited by Sp1 and Sp3 transcription factors. © 2003 European Society of Cardiology. Published by Elsevier B.V. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/195168
ISSN
2015 Impact Factor: 5.465
2015 SCImago Journal Rankings: 2.897
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorBrady, M-
dc.contributor.authorKoban, MU-
dc.contributor.authorDellow, KA-
dc.contributor.authorYacoub, M-
dc.contributor.authorBoheler, KR-
dc.contributor.authorFuller, SJ-
dc.date.accessioned2014-02-25T01:40:15Z-
dc.date.available2014-02-25T01:40:15Z-
dc.date.issued2003-
dc.identifier.citationCardiovascular Research, 2003, v. 60 n. 2, p. 347-354-
dc.identifier.issn0008-6363-
dc.identifier.urihttp://hdl.handle.net/10722/195168-
dc.description.abstractObjectives: The sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA) is essential to the removal of cytosolic calcium following cardiac contraction, and its abundance and activity are significantly altered during perinatal development and in failing myocardium. The objective of the current study was to identify cis regulatory elements and nuclear transcription factors responsible for transactivating SERCA2 gene expression in cardiomyocytes. Methods: Primary cultures of neonatal rat ventricular myocytes were transiently transfected with luciferase (LUX) reporter gene constructs containing deletions of the SERCA2 promoter or which harbored mutations in consensus Sp1 transcription factor binding sites. Cotransfection assays, electrophoretic mobility shift, and supershift assays were also performed to delineate the regulatory role of specific transcription factors. Results: We identified a putative AP-1-like element and a consensus Egr-1 binding site, but neither Egr-1 nor 12-O-tetradecanoylphorbol 13-acetate (TPA) significantly modified human SERCA2 promoter activity in vitro. Maximal activity of the SERCA2 promoter required the proximal 177 bp, and strong activation was observed with a 125-bp construct, within which an Sp1 site and a CAAT box were important. Mutation analysis also revealed the importance of two Sp1 sites between -125 and -200. Sp1 and Sp3 transcription factors were subsequently identified to bind to oligonucleotide probes corresponding to only the two most proximal Sp1 sites. Conclusions: These studies provide direct evidence that regulation of human SERCA2 gene expression in cardiomyocytes depends on transactivation events elicited by Sp1 and Sp3 transcription factors. © 2003 European Society of Cardiology. Published by Elsevier B.V. All rights reserved.-
dc.languageeng-
dc.relation.ispartofCardiovascular Research-
dc.titleSp1 and Sp3 transcription factors are required for trans-activation of the human SERCA2 promoter in cardiomyocytes-
dc.typeArticle-
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/S0008-6363(03)00529-7-
dc.identifier.pmid14613864-
dc.identifier.scopuseid_2-s2.0-0242627428-
dc.identifier.volume60-
dc.identifier.issue2-
dc.identifier.spage347-
dc.identifier.epage354-
dc.identifier.isiWOS:000186359500019-

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