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High Sensitivity Measurement of Transcription Factor-DNA Binding Affinities by Competitive Titration Using Fluorescence Microscopy
Published on: February 7, 2019
Different TRE-related elements are distinguished by sets of DNA-binding proteins with overlapping sequence
S E Smith1, A G Papavassiliou, D Bohmann
1European Molecular Biology Laboratory, Heidelberg, Germany.
Nucleic Acids Research
|April 11, 1993
Summary
Transcription factors bind to promoter elements like the TPA-responsive element (TRE). Different TRE-like sequences bind distinct subsets of transcription factors, including CREB and ATF proteins, influencing gene regulation.
Area of Science:
- Molecular Biology
- Gene Regulation
- Transcription Factor Binding
Background:
- The TPA-responsive element (TRE) is a key promoter element involved in gene regulation.
- Understanding the binding specificity of transcription factors to TRE and its variants is crucial for deciphering gene expression control.
Purpose of the Study:
- To compare the binding activities of different promoter elements with sequence similarity to the TRE.
- To identify transcription factors that bind to TRE-like elements in the collagenase, somatostatin, and c-jun promoters.
- To investigate the selectivity of these transcription factors for various TRE-like sequences.
Main Methods:
- Mobility-shift analyses were employed to assess DNA-protein interactions.
- Whole cell extracts were used to identify binding activities.
- Antibody 'supershift' assays were performed to identify protein components of the DNA-protein complexes.
Main Results:
- Specific DNA-binding activities were identified for TRE-like elements in the collagenase, somatostatin, and c-jun promoters.
- Different protein species exhibited varying degrees of selectivity for these TRE-like sequences.
- Components of the specific complexes were identified as CREB (cAMP response element-binding protein) and ATF (activating transcription factor) related products.
Conclusions:
- bZip protein dimers display differential tolerance to variations from the canonical TRE sequence.
- TRE-like promoter elements are distinguished by their ability to bind distinct subsets of related transcription factors.
- This differential binding contributes to the specificity of gene regulation by various promoter elements.
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