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PreprintCited by 6Cancer-related Molecular PathwaysProtein Structure and DynamicsDevelopmental Biology and Gene Regulation

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Evolution of affinity between p53 and MDM2 across the animal kingdom demonstrates high plasticity of motif-mediated interactions

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Mihalic, Filip, Åberg, Emma, Farkhondehkish, Pouria, Theys, Niels, Andersson, Eva, Jemth, Per·2023·Protein Science
DOI 10.1101/2023.01.26.525693PMC PMC10303687

Preprint — published as 10.1002/pro.4684 · Paper page

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Abstract

The interaction between the transcription factor p53 and the ubiquitin ligase MDM2 results in degradation of p53 and is well studied in cancer biology and drug development. Available sequence data suggest that both p53 and MDM2-family proteins are present across the animal kingdom. However, the interacting regions are missing in some animal groups, and it is not clear whether MDM2 interacts with, and regulates p53 in all species. We used phylogenetic analyses and biophysical measurements to examine the evolution of affinity between the interacting protein regions: a conserved 12-residue intrinsically disordered binding motif in the p53 transactivation domain (TAD) and the folded SWIB domain of MDM2. The affinity varied significantly across the animal kingdom. The p53TAD/MDM2 interaction among jawed vertebrates displayed high affinity, in particular for chicken and human proteins ( K D around 0.1 μM). The affinity of the bay mussel p53TAD/MDM2 complex was lower ( K D = 15 μM) and those from a placozoan, an arthropod and a jawless vertebrate were very low or non-detectable ( K D > 100 μM). Binding experiments with reconstructed ancestral p53TAD/MDM2 variants suggested that a micromolar affinity interaction was present in the ancestral bilaterian animal and was later enhanced in tetrapods while lost in other linages. The different evolutionary trajectories of p53TAD/MDM2 affinity during speciation demonstrate high plasticity of motif-mediated interactions and the potential for rapid adaptation of p53 regulation during times of change. Neutral drift in unconstrained disordered regions may underlie the plasticity and explain the observed low sequence conservation in transactivation domains such as p53TAD. Statement for broader audience: The protein p53 regulates central cellular processes including cell division and programmed cell death. p53 is regulated by another protein, MDM2, which binds to p53 and marks it for destruction. We measured the interaction between present-day and reconstructed ancient p53 and MDM2 proteins and found a range of binding strengths. Our findings suggest that rapid evolution of the p53/MDM2 interaction facilitates adaptation of p53 regulation during speciation.

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Metadata

Authors
Mihalic, F., Åberg, E., Farkhondehkish, P., Theys, N., Andersson, E., & Jemth, P.
Affiliations
Uppsala University (SE)
Corresponding author
Jemth, Per
ORCID
Venue
Protein Science
Publisher
openRxiv
Published
2023-01-26
Volume
32
Issue
7
Pages
e4684-e4684
ISSN
0961-8368 (ISSN-L)
DOI
10.1101/2023.01.26.525693
PMC
PMC10303687
OpenAlex
W4377195588
Cited by
6
References
1
Citations by year
2026: 2 · 2025: 1 · 2024: 1 · 2023: 2
FWCI
0.73
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Protein Science (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Uppsala University Publications (Uppsala University) (submittedVersion, other-oa)
Funders
  • Knut och Alice Wallenbergs Stiftelse (2015.0069)
  • Vetenskapsrådet (2020-04395)
Topics
  • Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
Keywords
p53, MDM2, p53 transactivation domain, protein-protein interaction affinity, intrinsically disordered regions, protein evolution, ancestral protein reconstruction, jawless fishes, placozoans, arthropods, neutral drift, p53 regulation
MeSH terms
Animals, Humans, Phylogeny, Protein Binding, Transcriptional Activation, Tumor Suppressor Protein p53, Protein Structure, Tertiary, Proto-Oncogene Proteins c-mdm2

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