title: HSP70 binding sites in the tumor suppressor protein p53.
abstract: Mutations within conserved regions of the tumor suppressor protein, p53, result in oncogenic forms of the protein with altered tertiary structures. In most cases, the mutant p53 proteins are selectively recognized and bound by members of the HSP70 family of molecular chaperones, but the binding site(s) in p53 for these chaperones have not been clearly defined. We have screened a library of overlapping biotinylated peptides, spanning the entire human p53 sequence, for binding to the HSP70 proteins, Hsc70 and DnaK. We show that most of the high affinity binding sites for these proteins map to secondary structure elements, particularly beta-strands, in the hydrophobic core of the central DNA binding domain, where the majority of oncogenic p53 mutations are found. Although peptides corresponding to the C-terminal region of p53 also contain potential binding sites, p53 proteins with C-terminal deletions are capable of binding to Hsc70, indicating that this region is not required for complex formation. We propose that mutations in the p53 protein alter the tertiary structure of the central DNA binding domain, thus exposing high affinity HSP70 binding sites that are cryptic in the wild-type molecule.
authors.affiliation: R. W. Johnson Pharmaceutical Research Institute, San Diego, California 92121, USA.
authors.forename: A M
authors.initials: AM
authors.lastname: Fourie
authors.forename: T R
authors.initials: TR
authors.lastname: Hupp
authors.forename: D P
authors.initials: DP
authors.lastname: Lane
authors.forename: B C
authors.initials: BC
authors.lastname: Sang
authors.forename: M S
authors.initials: MS
authors.lastname: Barbosa
authors.forename: J F
authors.initials: JF
authors.lastname: Sambrook
authors.forename: M J
authors.initials: MJ
authors.lastname: Gething
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D004247:DNA / Q000378:metabolism; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D016159:Tumor Suppressor Protein p53; D004247:DNA; C063842:dnaK protein, E coli