title: NMR spectroscopy reveals the solution dimerization interface of p53 core domains bound to their consensus DNA.
abstract: The p53 protein is a transcription factor that acts as the major tumor suppressor in mammals. The core DNA-binding domain is mutated in about 50% of all human tumors. The crystal structure of the core domain in complex with DNA illustrated how a single core domain specifically interacts with its DNA consensus site and how it is inactivated by mutation. However, no structural information for the tetrameric full-length p53-DNA complex is available. Here, we present novel experimental insight into the dimerization of two p53 core domains upon cooperative binding to consensus DNA in solution obtained by NMR. The NMR data show that the p53 core domain itself does not appear to undergo major conformational changes upon addition of DNA and elucidate the dimerization interface between two DNA-bound core domains, which includes the short H1 helix. A NMR-based model for the dimeric p53 core-DNA complex incorporates these data and allows the conclusion that the dimerization interface also forms the actual interface in the tetrameric p53-DNA complex. The significance of this interface is further corroborated by the finding that hot spot mutations map to the H1 helix, and by the binding of the putative p53 inhibitor 53BP2 to this region via one of its ankyrin repeats. Based on symmetry considerations it is proposed that tetrameric p53 might link non-contiguous DNA consensus sites in a sandwich-like manner generating DNA loops as observed for transcriptionally active p53 complexes.
authors.affiliation: Pharma Research, Roche Diagnostics GmbH, D-82372 Penzberg, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Klein
authors.forename: E
authors.initials: E
authors.lastname: Planker
authors.forename: T
authors.initials: T
authors.lastname: Diercks
authors.forename: H
authors.initials: H
authors.lastname: Kessler
authors.forename: K P
authors.initials: KP
authors.lastname: Künkele
authors.forename: K
authors.initials: K
authors.lastname: Lang
authors.forename: S
authors.initials: S
authors.lastname: Hansen
authors.forename: M
authors.initials: M
authors.lastname: Schwaiger
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D019281:Dimerization; D006801:Humans; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D011487:Protein Conformation; D020836:Protein Structure, Quaternary; D012996:Solutions; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000378:metabolism
chemical_list: D046911:Macromolecular Substances; D012996:Solutions; D016159:Tumor Suppressor Protein p53; D004247:DNA