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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References
Ashcroft M, Vousden KH . 1999 Oncogene 18: 7637–7643
Basi G, Schmid E, Maundrell K . 1993 Gene 123: 131–136
Baker SJ, Markowitz S, Fearon ER, Willson JK, Vogelstein B . 1990 Science 249: 912–915
Bischoff JR, Casso D, Beach D . 1992 Mol. Cell. Biol. 12: 1405–1411
Blattner C, Sparks A, Lane D . 1999 Mol. Cell. Biol. 19: 3704–3713
Brodsky MH, Nordstrom W, Tsang G, Kwan E, Rubin GM, Abrams JM . 2000 Cell 101: 103–113
Burch LR, Midgley CA, Currie RA, Lane DP, Hupp TR . 2000 FEBS Lett. 472: 93–98
Crook T, Vousden KH . 1992 EMBO J 11: 3935–3940
DiComo CJ, Prives C . 1998 Oncogene 16: 2527–2539
Diller L, Kassel J, Nelson CE, Gryka MA, Litwak G, Gebhardt M, Bressac B, Ozturk M, Baker SJ, Vogelstein B, et al. 1990 Mol. Cell. Biol. 10: 5772–5781
Epstein CB, Attiyeh EF, Hobson DA, Silver AL, Broach JR, Levine AJ . 1998 Oncogene 16: 2115–2122
Fuchs SY, Adler V, Buschmann T, Yin Z, Wu X, Jones SN, Ronai Z . 1998 Genes Dev. 12: 2658–2663
Funk WD, Pak DT, Karas RH, Wright WE, Shay JW . 1992 Mol. Cell. Biol. 12: 2866–2871
Grimm C, Kohli J, Murray J, Maundrell K . 1988 Proc. Natl. Acad. Sci. USA. 78: 5445–5449
Honda R, Tanaka H, Yasuda H . 1997 FEBS Lett. 420: 25–27
Ishioka C, Frebourg T, Yan YX, Vidal M, Friend SH, Schmidt S, Iggo R . 1993 Nat Genet. 5: 124–129
Juven-Gershon T, Shifman O, Unger T, Elkeles A, Haupt Y, Oren M . 1998 Mol. Cell. Biol. 18: 3974–3982
Kaelin Jr WG . 1999 Oncogene 18: 7701–7705
Kubbutat MHG, Jones SN, Vousden KH . 1997 Nature 387: 299–303
Kubbutat MH, Ludwig RL, Ashcroft M, Vousden KH . 1998 Mol Cell Biol 18: 5690–5698
Kussie PH, Gorina S, Marechal V, Elenbaas B, Moreau J, Levine AJ, Pavletich NP . 1996 Science 274: 948–953
Leng P, Brown DR, Shivakumar CV, Deb S, Deb SP . 1995 Oncogene 10: 1275–1282
Lin J, Chen J, Elenbaas B, Levine AJ . 1994 Genes Dev 8: 1235–1246
Matlashewsky G . 1999 Oncogene 18: 7618–7620
Maundrell K . 1993 Gene 123: 127–130
Mercer WE, Shields MT, Amin M, Sauve GJ, Appella E, Romano JW, Ullrich SJ . 1990 Proc. Natl. Acad. Sci. USA. 87: 6166–6170
Moreno S, Klar A, Nurse P . 1991 Methods Enzymol. 194: 795–823
Ollmann M, Young LM, DiComo CJ, Karim F, Belvin M, Robertson S, Whittaker K, Demsky M, Fisher WW, Buchman A, Duyk G, Friedman L, Prives C, Kopczynski C . 2000 Cell 101: 91–101
Riou G, Favre D, Jeannel J, Bourhis J, LeDoussal V, Orth G . 1990 Lancet. 335: 1171–1174
Rowan S, Ludwig RL, Haupt Y, Bates S, Lu X, Oren M, Vousden KH . 1996 EMBO J. 15: 827–838
Scharer E, Iggo R . 1992 Nucleic Acids Res. 20: 1539–1545
Scheffner M, Takahashi T, Huibregtse JM, Minna JD, Howley PM . 1992 J. Virol. 66: 5100–5105
Scheffner M, Huibregtse JM, Vierstra RD, Howley PM . 1993 Cell 75: 495–505
Slingerland JM, Jenkins JR, Benchimol S . 1993 EMBO J. 12: 1029–1037
Szankasi P, Heyer WD, Schuchert P, Kohli J . 1988 J. Mol. Biol. 204: 917–925
Tao W, Levine AJ . 1999 Proc. Natl. Acad. Sci. USA. 96: 3077–3080
Tarunina M, Jenkins JR . 1993 Oncogene 8: 3165–3173
Waddell S, Jenkins JR . 1995 Nucleic Acids Res. 23: 1836–1837
Waddell S, Jenkins JR . 1998 Oncogene 16: 1759–1765
Yonish-Rouach E, Resnitzky D, Lotem J, Sachs L, Kimchi A, Oren M . 1991 Nature 352: 345–347
Zeng X, Chen L, Jost CA, Maya R, Keller D, Wang X, Kaelin Jr WG, Oren M, Chen J, Lu H . 1999 Mol. Cell. Biol. 19: 3257–3266
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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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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DOI: https://doi.org/10.1038/sj.onc.1204702