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11:07
Biochemical Reconstitution of Steroid Receptor•Hsp90 Protein Complexes and Reactivation of Ligand Binding
Published on: September 21, 2011
Determinants for DNA-binding site recognition by the glucocorticoid receptor.
J Zilliacus1, A P Wright, U Norinder
1Center for Biotechnology, Karolinska Institute, NOVUM, Huddinge, Sweden.
The Journal of Biological Chemistry
|December 15, 1992
Summary
Glucocorticoid receptor DNA binding specificity is determined by three key amino acids. These residues, along with synergistic combinations, ensure accurate targeting of specific DNA sequences.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- The glucocorticoid receptor (GR) exhibits high specificity for glucocorticoid response elements (GREs).
- Discrimination between closely related DNA binding sites is crucial for gene regulation.
- Three amino acids in the GR DNA-binding domain's recognition helix are critical for DNA binding specificity.
Purpose of the Study:
- To elucidate the precise roles of three key amino acids in GR DNA binding specificity.
- To investigate how combinations of these residues influence binding to GREs and estrogen response elements (EREs).
- To develop models for amino acid-base pair associations in receptor-DNA interactions.
Main Methods:
- Construction and analysis of mutant GR DNA-binding domains with various amino acid substitutions.
- Systematic testing of mutant receptors for binding to GREs and EREs.
- Statistical analysis to model amino acid-base pair interactions and binding discrimination strategies.
Main Results:
- Specific amino acid residues, such as Val-443 in GR and Glu-439 in estrogen receptor (ER), mediate both positive contacts in cognate sites and negative contacts in non-cognate sites.
- Ser-440 in GR negatively impacts binding across all tested sites, with a stronger effect on EREs, enhancing discrimination.
- Synergistic interactions between amino acid combinations often lead to reduced binding at non-cognate sites.
Conclusions:
- Specific amino acid residues and their combinations are essential for the high fidelity of glucocorticoid receptor DNA binding.
- GR employs distinct strategies, including single residue contacts and synergistic effects, to discriminate between GREs and EREs.
- Understanding these molecular mechanisms provides insights into transcription factor specificity and gene regulation.
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