Related Experiment Videos
High affinity binding of MEF-2C correlates with DNA bending
D Meierhans1, M Sieber, R K Allemann
1Department of Chemistry, ETH-Zürich, Universitätstrasse 16, CH-8092 Zurich, Switzerland.
Nucleic Acids Research
|February 12, 1998
Summary
Myocyte enhancer factor-2 C (MEF-2C) transcription factors recognize DNA by sensing its inherent bendability. DNA sequence flexibility, not just base composition, is crucial for MEF-2C binding affinity.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Transcription factors regulate gene expression.
- Myocyte enhancer factor-2 (MEF-2) proteins cooperate with basic helix-loop-helix (bHLH) proteins.
- bHLH proteins have limited intrinsic DNA binding specificity.
Purpose of the Study:
- Investigate the in vitro DNA binding properties of MEF-2C.
- Determine the structural characteristics of DNA recognized by MEF-2C.
- Elucidate the role of DNA bendability in MEF-2C binding.
Main Methods:
- In vitro DNA binding assays (apparent dissociation constants).
- Electrophoretic mobility shift assays (EMSAs).
- Circular dichroism (CD) spectroscopy.
- Circular permutation assays for DNA bending.
Main Results:
- MEF-2C binds with high affinity to DNA sequences with MEF sites.
- DNA bendability is a principal structural feature recognized by MEF-2C.
- Reduced DNA bendability significantly decreased MEF-2C binding affinity.
Conclusions:
- MEF-2C binding specificity is influenced by the intrinsic bendability of the DNA sequence.
- DNA structural flexibility plays a critical role in transcription factor recognition.
- The study provides insights into the mechanism of MEF-2C-DNA interaction.