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PMID: 12023302 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cooperative action of Tbx2 and Nkx2.5 inhibits ANF expression in the atrioventricular canal: implications for cardiac chamber formation.

Genes & development ·Vol. 16 ·No. 10 ·2002-05-15 ·Pages 1234-46

Habets PE, Moorman AF, Clout DE, van Roon MA, Lingbeek M, van Lohuizen M, Campione M, Christoffels VM

Abstract

During heart development, chamber myocardium forms locally from the embryonic myocardium of the tubular heart. The atrial natriuretic factor (ANF) gene is specifically expressed in this developing chamber myocardium and is one of the first hallmarks of chamber formation. We investigated the regulatory mechanism underlying this selective expression. Transgenic analysis shows that a small fragment of the ANF gene is responsible for the developmental pattern of endogenous ANF gene expression. Furthermore, this fragment is able to repress cardiac troponin I (cTnI) promoter activity selectively in the embryonic myocardium of the atrioventricular canal (AVC). In vivo inactivation of a T-box factor (TBE)- or NK2-homeobox factor binding element (NKE) within the ANF fragment removed the repression in the AVC without affecting its chamber activity. The T-box family member Tbx2, encoding a transcriptional repressor, is expressed in the embryonic myocardium in a pattern mutually exclusive to ANF, thus suggesting a role in the suppression of ANF. Tbx2 formed a complex with Nkx2.5 on the ANF TBE-NKE, and was able to repress ANF promoter activity. Our data provide a potential mechanism for chamber-restricted gene activity in which the cooperative action of Tbx2 and Nkx2.5 inhibits expression in the AVC.

MeSH Terms
Animals Atrial Natriuretic Factor/antagonists & inhibitors,genetics,metabolism Binding Sites Blotting, Western Cell Cycle Proteins/genetics,metabolism Electrophoretic Mobility Shift Assay Embryonic and Fetal Development Enhancer Elements, Genetic/genetics Gene Expression Regulation, Developmental Heart Atria/embryology,metabolism Heart Ventricles/embryology,metabolism Homeobox Protein Nkx-2.5 Homeodomain Proteins/physiology In Situ Hybridization Mice Mice, Transgenic Mutagenesis, Site-Directed Myocardium/metabolism Plasmids Polymerase Chain Reaction Promoter Regions, Genetic T-Box Domain Proteins/physiology Transcription Factors Transcription, Genetic Troponin I/genetics,metabolism Xenopus Proteins
Chemicals
Cell Cycle Proteins Homeobox Protein Nkx-2.5 Homeodomain Proteins NKX2-5 protein, human Nkx2-5 protein, mouse T-Box Domain Protein 2 T-Box Domain Proteins Transcription Factors Troponin I Xenopus Proteins Atrial Natriuretic Factor
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Habets Petra E M H
Experimental and Molecular Cardiology Group, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
Moorman Antoon F M
Clout Danielle E W
van Roon Marian A
Lingbeek Merel
van Lohuizen Maarten
Campione Marina
Christoffels Vincent M
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.222902
Published
2002-05-15
Pages
1234-46
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC186286
Subset
IM
External Links
PubMed source DOI: 10.1101/gad.222902 PMC Full Text View journal details
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