Regulation of the cell-specific calcitonin/calcitonin gene-related peptide enhancer by USF and the Foxa2 forkhead protein

Tim J Viney; Thomas W Schmidt; William Gierasch; A Wahed Sattar; Ryan E Yaggie; Adisa Kuburas; John P Quinn; Judy M Coulson; Andrew F Russo

doi:10.1074/jbc.M406659200

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Regulation of the cell-specific calcitonin/calcitonin gene-related peptide enhancer by USF and the Foxa2 forkhead protein

Journal article

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Peer reviewed

Regulation of the cell-specific calcitonin/calcitonin gene-related peptide enhancer by USF and the Foxa2 forkhead protein

Tim J Viney, Thomas W Schmidt, William Gierasch, A Wahed Sattar, Ryan E Yaggie, Adisa Kuburas, John P Quinn, Judy M Coulson and Andrew F Russo

The Journal of biological chemistry, Vol.279(48), pp.49948-49955

11/26/2004

DOI: 10.1074/jbc.M406659200

PMID: 15385550

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Abstract

An 18-bp enhancer controls cell-specific expression of the calcitonin/calcitonin gene-related peptide gene. The enhancer is bound by a heterodimer of the bHLH-Zip protein USF-1 and -2 and a cell-specific factor from thyroid C cell lines. In this report we have identified the cell-specific factor as the forkhead protein Foxa2 (previously HNF-3beta). Binding of Foxa2 to the 18-bp enhancer was demonstrated using electrophoretic mobility shift assays. The cell-specific DNA-protein complex was selectively competed by a series of Foxa2 DNA binding sites, and the addition of Foxa2 antiserum supershifted the complex. Likewise, a complex similar to that seen with extracts from thyroid C cell lines was generated using an extract from heterologous cells expressing recombinant Foxa2. Interestingly, overexpression of Foxa2 activated the 18-bp enhancer in heterologous cells but only in the presence of the adjacent helix-loop-helix motif. Likewise, coexpression of USF proteins with Foxa2 yielded greater activation than by Foxa2 alone. Unexpectedly, Foxa2 overexpression repressed activity in the CA77 thyroid C cell line, suggesting that Foxa2 may interact with additional cofactors. The stimulatory role of Foxa2 at the calcitonin/calcitonin gene-related peptide gene enhancer was confirmed by short interfering RNA-mediated knockdown of Foxa2. As seen with Foxa2 overexpression, the effect of Foxa2 knockdown also required the adjacent helix-loop-helix motif. These results provide the first evidence for combinatorial control of gene expression by bHLH-Zip and forkhead proteins.

Hepatocyte Nuclear Factor 3-beta

Humans

Gene Expression Regulation - physiology

Rats

Calcitonin Gene-Related Peptide - genetics

Nuclear Proteins - metabolism

Calcitonin - genetics

Upstream Stimulatory Factors

DNA-Binding Proteins - metabolism

Transcription Factors - metabolism

Animals

Enhancer Elements, Genetic

Calcitonin Gene-Related Peptide - metabolism

Calcitonin - metabolism

Details

Title: Subtitle: Regulation of the cell-specific calcitonin/calcitonin gene-related peptide enhancer by USF and the Foxa2 forkhead protein
Creators: Tim J Viney - Department of Physiology and Biophysics, University of Iowa, Iowa City, Iowa 52242, USA
Thomas W Schmidt
William Gierasch
A Wahed Sattar
Ryan E Yaggie
Adisa Kuburas
John P Quinn
Judy M Coulson
Andrew F Russo
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.279(48), pp.49948-49955
DOI: 10.1074/jbc.M406659200
PMID: 15385550
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: HD 25969 / NICHD NIH HHS
Language: English
Date published: 11/26/2004
Academic Unit: Neurology; Molecular Physiology and Biophysics; Iowa Neuroscience Institute; Craniofacial Anomalies Research Center; Internal Medicine
Record Identifier: 9984020752902771

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