title: p53 derived from human tumour cell lines and containing distinct point mutations can be activated to bind its consensus target sequence.
abstract: Mutation of the p53 gene is one of the most common genetic lesions observed in human cancer. The p53 protein functions as a transcription factor, however it is still unresolved to what extend this property is related to its tumour suppressor activity. Since there is evidence that protein modifications as well as protein-protein interactions may regulate p53 function, we have studied p53 protein-DNA complex formation in nuclear extracts prepared from human tumour cell lines. In 13 different cell lines PAb421-induced DNA binding activity was compared to the level and conformation of the endogenous p53 protein. Surprisingly, sequence-specific p53 DNA binding activity was detected not only in cell lines that express wild-type p53, but also in seven cell lines which contain only mutant protein. Oligonucleotide competition analyses with various p53 target sequences and methylation interference experiments establish that wild-type and mutant p53 differ significantly in their sequence-specific interactions. Our analysis also provides evidence that the PAb1620 conformation is neither sufficient nor essential for DNA binding of endogenous p53 and that the cellular environment in addition to the specific point mutation may influence p53 DNA binding activity.
authors.affiliation: Department of Cellular and Moleculular Oncology, Masaryk Memorial Cancer Institute, Brno, Czech Republic.
authors.forename: D
authors.initials: D
authors.lastname: Niewolik
authors.forename: B
authors.initials: B
authors.lastname: Vojtesek
authors.forename: J
authors.initials: J
authors.lastname: Kovarik
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000378:metabolism*; D004249:DNA Damage; D006801:Humans; D016685:Mitomycin / Q000494:pharmacology; D008969:Molecular Sequence Data; D017354:Point Mutation; D011487:Protein Conformation; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000378:metabolism*
chemical_list: D016159:Tumor Suppressor Protein p53; D016685:Mitomycin; D004247:DNA