Cited by 371Cancer-related Molecular PathwaysHeat shock proteins researchCell death mechanisms and regulation

Design of a synthetic Mdm2-binding mini protein that activates the p53 response in vivo

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Böttger, Angelika, Böttger, Volker, Sparks, Alison, Liu, Wei-Li, Howard, Stephanie F., Lane, David P.·1997·Current Biology

Abstract

Background The transcriptional activation function of the p53 tumour suppressor protein is induced by DNA damage and results in growth arrest and/or apoptotic responses. A key component of this response is the dramatic rise in p53 protein concentration resulting from an increase in the protein's stability. Very recently, it has been suggested that interaction with the Mdm2 protein may target p53 for rapid degradation. We have designed a gene encoding a small protein that binds tightly to the p53-binding pocket on the Mdm2 protein. We have constructed the gene by cloning a phage display optimised Mdm2-binding peptide into the active-site loop of thioredoxin. Results When introduced into cells containing low levels of wild-type p53, this protein causes a striking accumulation of the endogenous p53 protein, activation of a p53-responsive reporter gene, and cell cycle arrest mimicking the effects seen in these cells after exposure to UV or ionising radiation. Microinjection of a monoclonal antibody to the p53-binding site on Mdm2 achieves a similar effect, establishing its specificity. Conclusions These results demonstrate that the p53 response is constitutively regulated in normal cells by Mdm2 and that disruption of the interaction alone is sufficient to stabilise the p53 protein and activate the p53 response. Our mini protein approach provides a powerful new method to activate p53 without causing DNA damage. More broadly, it establishes a powerful general method for determinin…

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Crossref · 10.1016/s0960-9822(06)00374-5

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Authors
Böttger, A., Böttger, V., Sparks, A., Liu, W.-L., Howard, S. F., & Lane, D. P.
Affiliations
University of Dundee (GB)
Corresponding author
Lane, David P.
Venue
Current Biology
Publisher
Elsevier BV
Published
1997-11
Volume
7
Issue
11
Pages
860-869
ISSN
0960-9822 (ISSN-L)
OpenAlex
W2041568308
Cited by
363
References
37
Citations by year
2024: 1 · 2023: 2 · 2022: 5 · 2021: 2 · 2020: 7 · 2019: 5 · 2018: 3 · 2017: 7 · 2016: 2 · 2015: 4 · 2014: 12 · 2013: 6 · 2012: 12
FWCI
11.86
Type
journal article
Language
en
Open access
Locations
  • Current Biology (publishedVersion)
  • PubMed (publishedVersion)
Funders
European Commission
Topics
  • Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
Keywords
p53, MDM2, p53-MDM2 interaction, protein-protein interaction disruption, thioredoxin, p53 stabilization, transcriptional activation, cell cycle arrest, phage display peptides, DNA damage response, tumour suppressor
MeSH terms
Amino Acid Sequence, Animals, Binding, Competitive, Cell Cycle, Cell Line, Humans, Male, Molecular Sequence Data, Nuclear Proteins, Prostate, Protein Binding, Proto-Oncogene Proteins, Recombinant Fusion Proteins, Thioredoxins, Transfection, Tumor Cells, Cultured, Protein Engineering, Transcriptional Activation, Tumor Suppressor Protein p53, Mice, Proto-Oncogene Proteins c-mdm2

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