title: Protein interactions at the carboxyl terminus of p53 result in the induction of its in vitro transactivation potential.
abstract: The tumor suppressor protein p53 is a transcription factor frequently inactivated in human cancers. We have studied the DNA binding potential and the transcriptional activity of p53 variants and p53 protein complexes in in vitro transcription assays. p53 specific transcription was measured via introduction of radioactive UTP into G-free cassette transcripts regulated by promoter sequences containing p53 response elements. Latent and activated p53 fractions were prepared from insect cells infected with p53 encoding baculoviruses by chromatography on heparin columns. p53 fractions distinguishable by their specific DNA binding activities and their recognition by monoclonal antibody PAb421 were obtained. Specific DNA binding and binding to PAb421 are mutually exclusive. The C-terminus of p53 can be phosphorylated by casein kinase II, protein kinase C and cyclin dependent kinases. The antibody PAb421 binds within the PKC phosphorylation site of p53 and is able to activate DNA binding of latent p53 in vitro. Activation of p53 by PAb421 also results in enhanced transactivation in vitro. Dephosphorylation of latent p53 with phosphatase 2A does not change these properties. This suggests that a conformational change in the carboxyl terminal domain of p53 controls the transactivation potential of p53.
authors.affiliation: Institute for Experimental Cancer Research, Tumor Biology Center, Freiburg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Mundt
authors.forename: T
authors.initials: T
authors.lastname: Hupp
authors.forename: M
authors.initials: M
authors.lastname: Fritsche
authors.forename: C
authors.initials: C
authors.lastname: Merkle
authors.forename: S
authors.initials: S
authors.lastname: Hansen
authors.forename: D
authors.initials: D
authors.lastname: Lane
authors.forename: B
authors.initials: B
authors.lastname: Groner
mesh_terms: D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011493:Protein Kinase C / Q000502:physiology; D018411:Spodoptera; D015533:Transcriptional Activation; D016159:Tumor Suppressor Protein p53 / Q000502:physiology*
chemical_list: D016159:Tumor Suppressor Protein p53; D004247:DNA; D011493:Protein Kinase C