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Updated: Aug 16, 2026
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Detecting the Ligand-binding Domain Dimerization Activity of Estrogen Receptor Alpha Using the Mammalian Two-Hybrid Assay
Published on: December 19, 2018
DNA recognition by the oestrogen receptor: from solution to the crystal
J W Schwabe1, L Chapman, J T Finch
1MRC Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, UK.
Structure (London, England : 1993)
|November 15, 1993
Summary
The estrogen receptor DNA-binding domain (ERDBD) binds DNA cooperatively. Specific disordered regions become ordered upon DNA binding, facilitating dimer formation and enhancing sequence recognition.
Area of Science:
- Molecular Biology
- Structural Biology
- Genetics
Background:
- Steroid/nuclear hormone receptors are transcription factors regulating gene expression.
- They bind DNA response elements as dimers, recognizing specific sequences.
- A conserved DNA-binding domain mediates this recognition.
Purpose of the Study:
- To characterize the DNA-binding properties of the estrogen receptor DNA-binding domain (ERDBD).
- To determine the refined NMR structure of the ERDBD.
- To elucidate the mechanism of ERDBD DNA recognition and binding cooperativity.
Main Methods:
- Nuclear Magnetic Resonance (NMR) spectroscopy to determine the solution structure of ERDBD.
- Analysis of DNA-binding properties and cooperativity.
- Comparison with existing X-ray crystallographic data.
Main Results:
- The isolated ERDBD is monomeric in solution.
- Two ERDBD molecules bind DNA cooperatively to specific sequences.
- Disordered regions in the solution structure become ordered upon DNA binding, forming a dimer interface.
Conclusions:
- Cooperative DNA binding by ERDBD is crucial for its function.
- Ordered internal residues mediate DNA contact and dimer formation.
- These findings explain how ERDBD achieves specific DNA recognition through cooperative dimerization.
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