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We have developed a functional "no-hybrids" screen in the fission yeast Schizosaccharomyces pombe based on the transcription transactivator activity of human p53. The screen can be used to identify antagonizers and modulators of p53 activity. Expression of functional full-length human p53 is conditionally lethal to the screen reporter strains. Co-expression of specific inhibitory proteins promotes cell survival and growth. We have validated the "no-hybrids" system by (a) successful modeling of human wild-type p53 interaction with SV40 large T antigen, Mdm2 and a panel of tumor-derived human p53 mutants, (b) demonstrating the screening system's efficiency through identification of a dominant negative fragment of p53 itself in a library screen context and (c) using Drosophila p53 to demonstrate that the system can detect evolutionarily distant p53 homologues based on their transactivator activity. The "no-hybrids" screen will be of utility in searches for p53 function-modulators of both cellular and viral origin.

Original publication

DOI

10.1038/sj.onc.1204702

Type

Journal article

Journal

Oncogene

Publication Date

20/09/2001

Volume

20

Pages

6001 - 6008

Keywords

Animals, Cell Division, Drosophila melanogaster, Gene Library, Genes, Reporter, Humans, Insect Proteins, Mutation, Schizosaccharomyces, Temperature, Trans-Activators, Tumor Suppressor Protein p53, Two-Hybrid System Techniques