Related Experiment Video
Updated: Mar 31, 2026

High Sensitivity Measurement of Transcription Factor-DNA Binding Affinities by Competitive Titration Using Fluorescence Microscopy
Published on: February 7, 2019
The p53 tetramer shows an induced-fit interaction of the C-terminal domain with the DNA-binding domain
M D'Abramo1, N Bešker2, A Desideri3
1Dipartimento di Chimica, Sapienza University of Rome, Rome, Italy.
Abstract:
The Trp53 gene is the most frequently mutated gene in all human cancers. Its protein product p53 is a very powerful transcription factor that can activate different biochemical pathways and affect the regulation of metabolism, senescence, DNA damage response, cell cycle and cell death. The understanding of its function at the molecular level could be of pivotal relevance for therapy. Investigation of long-range intra- and interdomain communications in the p53 tetramer-DNA complex was performed by means of an atomistic model that included the tetramerization helices in the C-terminal domain, the DNA-binding domains and a consensus DNA-binding site of 18 base pairs. Nonsymmetric dynamics are illustrated in the four DNA-binding domains, with loop L1 switching from inward to outward conformations with respect to the DNA major groove. Direct intra- and intermonomeric long-range communications between the tetramerization and DNA-binding domains are noted. These long-distance conformational changes link the C terminus with the DNA-binding domain and provide a biophysical rationale for the reported functional regulation of the p53 C-terminal region. A fine characterization of the DNA deformation caused by p53 binding is obtained, with 'static' deformations always present and measured by the slide parameter in the central thymine-adenine base pairs; we also detect 'dynamic' deformations switched on and off by particular p53 tetrameric conformations and measured by the roll and twist parameters in the same base pairs. These different conformations can indeed modulate the electrostatic potential isosurfaces of the whole p53-DNA complex. These results provide a molecular/biophysical understanding of the evident role of the C terminus in post-translational modification that regulates the transcriptional function of p53. Furthermore, the unstructured C terminus is able to facilitate contacts between the core DNA-binding domains of the tetramer.
Insights
The p53 protein
Area of Science:
- Molecular Biology
- Biophysics
- Genetics
Background:
- The Trp53 gene is frequently mutated in human cancers.
- The p53 protein is a crucial transcription factor regulating key cellular processes.
- Understanding p53's molecular function is vital for cancer therapy.
Purpose of the Study:
- Investigate long-range communication within the p53 tetramer-DNA complex.
- Elucidate the role of the C-terminal domain in p53 regulation.
- Characterize p53-induced DNA deformations.
Main Methods:
- Atomistic modeling of the p53 tetramer-DNA complex.
- Analysis of nonsymmetric dynamics in DNA-binding domains.
- Measurement of DNA deformation parameters (slide, roll, twist).
Main Results:
- Identified direct long-range communications between p53 domains.
- Observed dynamic conformational changes in DNA-binding loops.
- Characterized both static and dynamic DNA deformations induced by p53.
- Demonstrated modulation of the complex's electrostatic potential by p53 conformations.
Conclusions:
- Provided a biophysical basis for C-terminal regulation of p53 transcriptional activity.
- Highlighted the role of the C terminus in facilitating DNA-binding domain interactions.
- Offered molecular insights into p53's function in cancer biology.
Related Concept Videos
Abnormal Proliferation
DNA Damage can Stall the Cell Cycle
DNA Damage Can Stall the Cell Cycle
Single-Strand DNA Binding Proteins
Covalently Linked Protein Regulators
These groups modify specific amino acids in a protein....
Restarting Stalled Replication Forks

