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A complex intronic enhancer regulates expression of the CFTR gene by direct interaction with the promoter.

Publication ,  Journal Article
Ott, CJ; Suszko, M; Blackledge, NP; Wright, JE; Crawford, GE; Harris, A
Published in: J Cell Mol Med
April 2009

Genes can maintain spatiotemporal expression patterns by long-range interactions between cis-acting elements. The cystic fibrosis transmembrane conductance regulator gene (CFTR) is expressed primarily in epithelial cells. An element located within a DNase I-hypersensitive site (DHS) 10 kb into the first intron was previously shown to augment CFTR promoter activity in a tissue-specific manner. Here, we reveal the mechanism by which this element influences CFTR transcription. We employed a high-resolution method of mapping DHS using tiled microarrays to accurately locate the intron 1 DHS. Transfection of promoter-reporter constructs demonstrated that the element displays classical tissue-specific enhancer properties and can independently recruit factors necessary for transcription initiation. In vitro DNase I footprinting analysis identified a protected region that corresponds to a conserved, predicted binding site for hepatocyte nuclear factor 1 (HNF1). We demonstrate by electromobility shift assays (EMSA) and chromatin immunoprecipitation (ChIP) that HNF1 binds to this element both in vitro and in vivo. Moreover, using chromosome conformation capture (3C) analysis, we show that this element interacts with the CFTR promoter in CFTR-expressing cells. These data provide the first insight into the three- dimensional (3D) structure of the CFTR locus and confirm the contribution of intronic cis-acting elements to the regulation of CFTR gene expression.

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Published In

J Cell Mol Med

DOI

EISSN

1582-4934

Publication Date

April 2009

Volume

13

Issue

4

Start / End Page

680 / 692

Location

England

Related Subject Headings

  • Transfection
  • Transcription, Genetic
  • Transcription Factors
  • Protein Binding
  • Promoter Regions, Genetic
  • Organ Specificity
  • Molecular Sequence Data
  • Introns
  • Humans
  • Hepatocyte Nuclear Factor 1
 

Citation

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Ott, C. J., Suszko, M., Blackledge, N. P., Wright, J. E., Crawford, G. E., & Harris, A. (2009). A complex intronic enhancer regulates expression of the CFTR gene by direct interaction with the promoter. J Cell Mol Med, 13(4), 680–692. https://doi.org/10.1111/j.1582-4934.2008.00621.x
Ott, Christopher J., Magdalena Suszko, Neil P. Blackledge, Jane E. Wright, Gregory E. Crawford, and Ann Harris. “A complex intronic enhancer regulates expression of the CFTR gene by direct interaction with the promoter.J Cell Mol Med 13, no. 4 (April 2009): 680–92. https://doi.org/10.1111/j.1582-4934.2008.00621.x.
Ott CJ, Suszko M, Blackledge NP, Wright JE, Crawford GE, Harris A. A complex intronic enhancer regulates expression of the CFTR gene by direct interaction with the promoter. J Cell Mol Med. 2009 Apr;13(4):680–92.
Ott, Christopher J., et al. “A complex intronic enhancer regulates expression of the CFTR gene by direct interaction with the promoter.J Cell Mol Med, vol. 13, no. 4, Apr. 2009, pp. 680–92. Pubmed, doi:10.1111/j.1582-4934.2008.00621.x.
Ott CJ, Suszko M, Blackledge NP, Wright JE, Crawford GE, Harris A. A complex intronic enhancer regulates expression of the CFTR gene by direct interaction with the promoter. J Cell Mol Med. 2009 Apr;13(4):680–692.
Journal cover image

Published In

J Cell Mol Med

DOI

EISSN

1582-4934

Publication Date

April 2009

Volume

13

Issue

4

Start / End Page

680 / 692

Location

England

Related Subject Headings

  • Transfection
  • Transcription, Genetic
  • Transcription Factors
  • Protein Binding
  • Promoter Regions, Genetic
  • Organ Specificity
  • Molecular Sequence Data
  • Introns
  • Humans
  • Hepatocyte Nuclear Factor 1