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Binding of glucocorticoid receptors to model DNA response elements
M Benore-Parsons1, J Liebman, L P Wennogle
1Research Division, CIBA-GEIGY Corporation, Summit, New Jersey 07901.
Journal of Cellular Biochemistry
|December 1, 1991
Summary
Optimal glucocorticoid receptor binding requires approximately 45 base pairs of DNA, including flanking sequences. Multiple receptor binding sites enhance this interaction, impacting gene transcription.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Glucocorticoids regulate gene expression through the glucocorticoid receptor (GR).
- The consensus glucocorticoid response element (GRE) is a specific DNA sequence recognized by GR.
- The role of DNA sequences flanking the core GRE in GR binding and transcriptional regulation is not fully understood.
Purpose of the Study:
- To investigate the optimal DNA sequence length for glucocorticoid receptor binding.
- To determine the influence of flanking sequences on GR-DNA interactions.
- To explore the effect of multiple GREs on GR binding affinity and transcriptional synergy.
Main Methods:
- Synthesis of DNA oligomers containing a consensus 15-bp GRE with varying flanking sequences.
- In vitro binding assays using a reconstituted system with purified glucocorticoid receptor.
- Competition assays to assess the binding affinity of different DNA sequences.
Main Results:
- A minimal DNA length of approximately 45 base pairs was found to be necessary for optimal GR binding.
- DNA sequences with multiple GREs exhibited enhanced competition for GR binding.
- Flanking sequences significantly contribute to optimizing GR binding interactions.
Conclusions:
- Optimal glucocorticoid receptor binding to DNA is dependent on both the core GRE and sufficient flanking sequences.
- Synergistic effects of multiple GREs play a crucial role in glucocorticoid-mediated transcriptional control.
- The study confirms the importance of DNA sequence context in receptor-DNA interactions and gene regulation.