Reactivation of mutant p53 by a one-hybrid adaptor protein.
The most frequent genetic alteration in cancer is a mutation of p53. In most cases, this leads to a sharp increase of the p53 protein levels but abolishes p53's function as an activator of transcription. To correct this defect, wild-type p53 is being reintroduced into tumor cells through gene therapy vectors, thereby inducing cell death. However, this effect is not necessarily specific for tumor cells. Furthermore, mutant p53 in tumor cells trans-dominantly impairs the function of wild-type p53. As an approach to overcome these obstacles, we have developed an adaptor protein that reactivates mutant p53 rather than stimulating transcription on its own. The DNA binding and tetramerizing portions of the p53-homologue p73 were fused to the oligomerization domain of p53. This chimera binds to the DNA of p53-responsive promoters through the p73-derived portions, and it binds to mutant p53 by the p53-derived oligomerization domain. Through this one-hybrid system, mutant p53 is re-enabled to activate transcription. When the adaptor was expressed in tumor cells that contain mutant p53, expression of p53-responsive genes was activated, and growth was inhibited. No such effects were observed in cells that contain wild-type p53 or no p53 at all. When the adaptor was expressed through an adenovirus vector, tumor cells containing mutant p53 were specifically induced to undergo apoptosis. This strategy can turn mutant p53 into an inhibitor of tumor cell growth and might enable gene therapy to eliminate cancer cells with specificity.
Roth
Judith
J
Abteilung für Gastroenterologie und Stoffwechsel, Klinikum der Philipps-Universität Marburg, Baldingerstrasse, 35043 Marburg, Germany.
Lenz-Bauer
Claudia
C
Contente
Ana
A
Löhr
Kristina
K
Koch
Philipp
P
Bernard
Sandra
S
Dobbelstein
Matthias
M
D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D016147:Genes, Tumor Suppressor; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation; D014162:Transfection; D014407:Tumor Cells, Cultured; D000072160:Tumor Protein p73; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D025521:Tumor Suppressor Proteins
D033942:Adaptor Proteins, Vesicular Transport; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; C000606460:TP73 protein, human; D000072160:Tumor Protein p73; D016159:Tumor Suppressor Protein p53; D025521:Tumor Suppressor Proteins; D004247:DNA