| Design of a synthetic Mdm2-binding mini protein that activates the p53 response in vivo. | |
MedLine Citation:
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PMID: 9382809 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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 determining the biological consequences of the specific disruption of protein-protein interactions in cells. |
Authors:
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A Böttger; V Böttger; A Sparks; W L Liu; S F Howard; D P Lane |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Current biology : CB Volume: 7 ISSN: 0960-9822 ISO Abbreviation: Curr. Biol. Publication Date: 1997 Nov |
Date Detail:
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Created Date: 1998-02-25 Completed Date: 1998-02-25 Revised Date: 2008-11-21 |
Medline Journal Info:
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Nlm Unique ID: 9107782 Medline TA: Curr Biol Country: ENGLAND |
Other Details:
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Languages: eng Pagination: 860-9 Citation Subset: IM |
Affiliation:
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Cancer Research Campaign Laboratories, University of Dundee, UK. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Animals Binding, Competitive Cell Cycle / drug effects, genetics Cell Line Humans Male Mice Molecular Sequence Data Nuclear Proteins* Prostate / cytology Protein Binding / drug effects, genetics Protein Engineering / methods* Proto-Oncogene Proteins / antagonists & inhibitors, chemical synthesis*, genetics, metabolism, pharmacology* Proto-Oncogene Proteins c-mdm2 Recombinant Fusion Proteins / biosynthesis, chemical synthesis, physiology Thioredoxins / genetics Transcriptional Activation Transfection Tumor Cells, Cultured Tumor Suppressor Protein p53 / antagonists & inhibitors, biosynthesis*, genetics, physiology |
| Chemical | |
Reg. No./Substance:
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0/Nuclear Proteins; 0/Proto-Oncogene Proteins; 0/Recombinant Fusion Proteins; 0/Tumor Suppressor Protein p53; 52500-60-4/Thioredoxins; EC 6.3.2.19/MDM2 protein, human; EC 6.3.2.19/Mdm2 protein, mouse; EC 6.3.2.19/Proto-Oncogene Proteins c-mdm2 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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