Amifostine (WR2721) restores transcriptional activity of specific p53 mutant proteins in a yeast functional assay.
Many p53 mutants found in human cancer have an altered ability to bind DNA and transactivate gene expression. Re-expression of functional p53 in cells in which the endogenous TP53 gene is inactivated has been demonstrated to restore a non-tumorigenic phenotype. Pharmacological modulation of p53 mutant conformation may therefore represent a mechanism to reactivate p53 function and consequently improve response to radio- and chemotherapy. We have recently reported that the radio- and chemoprotector Amifostine (WR2721, Ethyol) activates wild-type p53 in cultured mammalian cells. In the present study, we have used a yeast functional assay to investigate the effect of WR2721 on the transcriptional activity of p53. WR2721 restored this activity in a temperature-sensitive mutant V272M (valine to methionine at codon 272) expressed at the non-permissive temperature and it also partially restored the transcriptional activity of several other conformationally flexible p53 mutants. The results indicate that the yeast functional assay may be used to identify compounds that modulate p53 activity, with potential therapeutic implications.
Maurici
D
D
Group of Molecular Carcinogenesis, International Agency for Research on Cancer, 150 Cours Albert Thomas, 69372 Lyon, France.
Monti
P
P
Campomenosi
P
P
North
S
S
Frebourg
T
T
Fronza
G
G
Hainaut
P
P
D004999:Amifostine / Q000494:pharmacology*; D019943:Amino Acid Substitution; D000818:Animals; D003001:Cloning, Molecular; D003062:Codon; D004938:Esophageal Neoplasms / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008322:Mammals; D008715:Methionine; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D011073:Polyamines / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011837:Radiation-Protective Agents / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000254:growth & development; D013696:Temperature; D014158:Transcription, Genetic / Q000187:drug effects*; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014633:Valine
D003062:Codon; D011073:Polyamines; D011837:Radiation-Protective Agents; D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53; D008715:Methionine; D014633:Valine; D004999:Amifostine