Cell-specific activation of the atrial natriuretic factor promoter by PITX2 and MEF2A

Rafael Toro; Irfan Saadi; Adisa Kuburas; Mona Nemer; Andrew F Russo

doi:10.1074/jbc.M404802200

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Cell-specific activation of the atrial natriuretic factor promoter by PITX2 and MEF2A

Journal article

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Peer reviewed

Cell-specific activation of the atrial natriuretic factor promoter by PITX2 and MEF2A

Rafael Toro, Irfan Saadi, Adisa Kuburas, Mona Nemer and Andrew F Russo

The Journal of biological chemistry, Vol.279(50), pp.52087-52094

12/10/2004

DOI: 10.1074/jbc.M404802200

PMID: 15466416

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Abstract

The PITX2 homeodomain protein is mutated in patients with Axenfeld-Rieger syndrome and is involved in the development of multiple organ systems, including the heart. We have examined the interaction of PITX2 isoforms with myocyte-enhancing factor 2A (MEF2A), which is a known regulator of cardiac development. A direct interaction between PITX2a and MEF2A was demonstrated using yeast two-hybrid and GST pull-down assays. To study the functional significance of this interaction, we used the atrial natriuretic factor (ANF) promoter. Coexpression of MEF2A and PITX2a or Pitx2c resulted in a strong synergistic activation of the ANF promoter in LS8 oral epithelial cells but not in other cell lines (NIH/3T3, Chinese hamster ovary, or C2C12). The synergism was dependent on promoter context, because it required MEF2 binding sites and was not seen with two other PITX2 target promoters. DNA binding by MEF2A was required but not sufficient for synergism. Upstream activators of p38 MAP kinases, MKK3 and MKK6, increased PITX2a and Pitx2c activity to yield up to 90-fold activation of the ANF promoter in LS8 cells. Because Axenfeld-Rieger syndrome is autosomal dominant and affects development of the oral epithelium, we tested one of the known PITX2 mutants. The PITX2a-K88E mutant protein suppressed wild type PITX2a synergism with MEF2A. These results demonstrate a promoter- and cell-specific functional interaction between PITX2 and MEF2A and suggest the possibility of coordinate control by these factors in the oral epithelium.

NIH 3T3 Cells

Epithelial Cells - metabolism

Homeodomain Proteins - metabolism

Humans

DNA-Binding Proteins - metabolism

Myoblasts - metabolism

Atrial Natriuretic Factor - genetics

p38 Mitogen-Activated Protein Kinases - metabolism

Nuclear Proteins - genetics

MEF2 Transcription Factors

CHO Cells

Recombinant Proteins - metabolism

Cell Line

Promoter Regions, Genetic

Cricetinae

Gene Expression Regulation

Nuclear Proteins - metabolism

Recombinant Proteins - genetics

Binding Sites - genetics

Myogenic Regulatory Factors

Transcription Factors - genetics

DNA-Binding Proteins - genetics

Homeodomain Proteins - genetics

Transcription Factors - metabolism

Two-Hybrid System Techniques

Animals

MADS Domain Proteins

Mice

In Vitro Techniques

Details

Title: Subtitle: Cell-specific activation of the atrial natriuretic factor promoter by PITX2 and MEF2A
Creators: Rafael Toro - Genetics Program, University of Iowa, Iowa City, Iowa 52242, USA
Irfan Saadi
Adisa Kuburas
Mona Nemer
Andrew F Russo
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.279(50), pp.52087-52094
DOI: 10.1074/jbc.M404802200
PMID: 15466416
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: E13076 / PHS HHS
Language: English
Date published: 12/10/2004
Academic Unit: Neurology; Molecular Physiology and Biophysics; Iowa Neuroscience Institute; Craniofacial Anomalies Research Center
Record Identifier: 9984019536802771

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