Cited by 95Protein Structure and DynamicsCancer-related Molecular PathwaysAdvanced NMR Techniques and Applications

Core domain interactions in full-length p53 in solution

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Veprintsev, Dmitry B., Freund, Stefan M. V., Andreeva, Antonina, Rutledge, Stacey E., Tidow, Henning, Cañadillas, José Manuel Pérez, Blair, Caroline M., Fersht, Alan R.·2006·Proceedings of the National Academy of Sciences

Abstract

The tumor suppressor p53 consists of four 393-residue chains, each of which has two natively unfolded (N- and C-terminal) and two folded (core and tetramerization) domains. Their structural organization is poorly characterized as the protein tends to aggregate, has defied crystallization, and is at the limits of NMR studies. We first stabilized the protein by mutation to make it more suitable for extended study and then acquired NMR spectra on full-length protein and various combinations of shorter domain constructs. The NMR spectrum ( 15 N, 1 H transverse relaxation optimized spectroscopy) of full-length p53 was close to that expected from the sum of the spectra of isolated individual domains. However, patterns of changes in chemical shifts revealed unexpected interactions between the core domains. We used the NMR data as constraints in docking algorithms and found a previously uncharacterized self-complementary surface for the association of core domains into dimers within the tetrameric complex. Binding to DNA requires about a 70° rotation to break those subunit interactions and form the known protein:protein interface in the p53–DNA complex. We verified the interactions by the effects of mutation on DNA binding. Spectroscopic, biophysical, and mutational data conspired to give a picture of the p53 tetramer as a dimer of loosely tethered core dimers of appropriate symmetry to be poised to bind target DNA.

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Crossref · 10.1073/pnas.0511130103

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  • Typically, p53 binds to target genes as a tetramer comprising two dimers, each binding a decameric half-site with the consensus sequence RRRCWWGYYY.

    “Typically, p53 binds to the target genes as a tetramer, which comprises two dimers that each binds a decameric half-site with the consensus sequence RRRCWWGYYY (R = A/G, W = A/T, Y = C/T).7, 8, 9, 10”

    From Census and evaluation of p53 target genes (2017) · cites this paper as [10]

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Authors
Veprintsev, D. B., Freund, S. M. V., Andreeva, A., Rutledge, S. E., Tidow, H., Cañadillas, J. M. P., Blair, C. M., & Fersht, A. R.
Affiliations
  • Medical Research Council (GB)
  • Medical Research Council Centre for Protein Engineering, Medical Research Council Centre, Hills Road, Cambridge CB2 2QH, United Kingdom
Corresponding author
Fersht, Alan R.
Venue
Proceedings of the National Academy of Sciences
Published
2006-02-06
Volume
103
Issue
7
Pages
2115-2119
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
OpenAlex
W2118505770
Cited by
95
References
35
Citations by year
2026: 1 · 2025: 4 · 2023: 1 · 2022: 2 · 2021: 2 · 2020: 1 · 2019: 2 · 2018: 1 · 2017: 4 · 2016: 2 · 2015: 4 · 2014: 2 · 2013: 3 · 2012: 9
FWCI
5.58
Type
journal article
Language
en
Open access
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
  • Physical Sciences › Chemistry › Spectroscopy › Advanced NMR Techniques and Applications
Keywords
p53, tumor suppressor proteins, protein domain interactions, core domain, tetramerization domain, protein dimerization, protein-protein interaction, p53 DNA binding, protein aggregation, chemical shift perturbation, protein solution structure
MeSH terms
Crystallography, DNA, Humans, Solutions, Tumor Suppressor Protein p53, Mutagenesis, Protein Structure, Tertiary, Dimerization

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