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A‘no-hybrids’ screen for functional antagonizers of human p53 transactivator function: dominant negativity in fission yeast

Oncogene volume 20, pages 6001–6008 (2001)Cite this article

Abstract

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.

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Acknowledgements

This work was carried out at the Marie Curie Research Institute, Oxted, UK. We are grateful to S Forsburg, D Hughes, DP Lane, JD Milner, K Vousden for kindly providing reagents used in this study. T Proikas-Cezanne was a recipient of a post-doctoral fellowship of the Deutsche Forschungsgemeinschaft (Pr 541-2).

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Authors and Affiliations

  1. Department of Brain and Cognitive Sciences, Department of Biology, Center for Learning and Memory, Massachusetts Institute of Technology, Cambridge, MA 02139, Massachusetts, USA

    Scott Waddell

  2. Fels Institute for Cancer Research and Molecular Biology, Temple University, School of Medicine, 3307 N. Broad Street, Philadelphia, PA 19140, Pennsylvania, USA

    John R Jenkins & Tassula Proikas-Cezanne

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  1. Scott Waddell
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Correspondence to Tassula Proikas-Cezanne.

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Waddell, S., Jenkins, J. & Proikas-Cezanne, T. A‘no-hybrids’ screen for functional antagonizers of human p53 transactivator function: dominant negativity in fission yeast. Oncogene 20, 6001–6008 (2001). https://doi.org/10.1038/sj.onc.1204702

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