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Chromatin Immunoprecipitation from Human Embryonic Stem Cells
Published on: July 22, 2008
Transcription factor cCP2 controls gene expression in chicken embryonic stem cells
Hervé Acloque1, Anne Mey, Anne Marie Birot
1Laboratoire de Biologie Moléculaire de la Cellule, CNRS-INRA UMR5161, Ecole Normale Supérieure de Lyon, IFR128 BioSciences Lyon-Gerland, 46 Allée d'Italie, 69364 Lyon Cedex 07, France.
Nucleic Acids Research
|April 27, 2004
Summary
Chicken embryonic stem cell (CES) genes cENS-1/cERNI are regulated by a specific promoter region. Transcription factor CP2 binds to this region, proving crucial for early embryonic development gene activation.
Area of Science:
- Developmental Biology
- Gene Regulation
- Molecular Biology
Background:
- cENS-1/cERNI genes are expressed early in chicken embryonic development and in pluripotent chicken embryonic stem (CES) cells.
- A specific promoter region active in CES cells, but not differentiated cells, has been identified for cENS-1/cERNI.
Purpose of the Study:
- To elucidate the molecular mechanisms regulating the cENS-1/cERNI promoter.
- To identify cis-acting elements responsible for promoter activity in CES and differentiated cells.
Main Methods:
- Analysis of cis-acting elements within the cENS-1/cERNI promoter.
- Site-directed mutagenesis of identified regulatory regions.
- Supershift experiments to identify DNA-binding proteins.
Main Results:
- A novel enhancer sequence, the B region (5'-CAAG TCCAGG CAAG-3'), was identified with strong activity in both CES and differentiated cells.
- Mutation of the B region significantly reduced cENS-1/cERNI promoter activity in CES cells.
- Supershift assays confirmed the binding of a protein complex containing the transcription factor CP2 to the B region.
Conclusions:
- The B region is essential for activating the cENS-1/cERNI promoter in CES cells.
- A nuclear factor from the CP2 transcription factor family is crucial for cENS-1/cERNI promoter activation.
- The expression pattern of cCP2 in early chicken embryos suggests its essential role in early embryonic gene expression, mirroring cENS-1/cERNI expression.
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