title: Specific and complex interactions of murine p53 with DNA.
abstract: Biologically active mutant p53 from Balb/c mouse tumor cells (Meth A) was analysed for its specific interaction with DNA. Restricted phage lambda DNA, representing DNA of high complexity with regard to sequence and secondary structure, was used to probe for such an activity in a target-bound DNA-binding assay, using doubly immunopurified p53. A single lambda DNA fragment was specifically retained with very high affinity (KD = 10(-10) M). Specific DNA binding was shown to be an intrinsic property of p53, as it could be blocked with p53-specific monoclonal antibodies PAb122 and PAb421. The characteristics of the DNA binding of p53 to this lambda DNA fragment, as well as the structural properties of this fragment, suggested the possibility that p53 might be able to interact with nuclear matrix attachment region (MAR) DNA. Indeed, established genomic MAR elements were specifically bound by Meth A p53, whereas no binding was observed to an AT-rich control DNA. The interaction of p53 with MAR elements in vitro is compatible with the idea that p53 in vivo is involved in the regulation of replication and/or expression of cellular DNA. Complex DNA interactions were not restricted to mutant p53 from Meth A cells. Mutant p53 of a different conformational phenotype (PAb246+ 'wild-type' as opposed to PAb246- 'mutant' for p53 from Meth A cells) from minimally transformed T3T3 cells, as well as genotypic wild-type p53 expressed by a recombinant baculovirus in insect cells, exhibited similar DNA-binding properties.
authors.affiliation: Heinrich-Pette-Institut für Experimentelle Virologie und Immunologie, Universität Hamburg, Germany.
authors.forename: S N
authors.initials: SN
authors.lastname: Weissker
authors.forename: B F
authors.initials: BF
authors.lastname: Müller
authors.forename: A
authors.initials: A
authors.lastname: Homfeld
authors.forename: W
authors.initials: W
authors.lastname: Deppert
mesh_terms: D000818:Animals; D004247:DNA / Q000378:metabolism*; D016158:Genes, p53; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D015530:Nuclear Matrix / Q000378:metabolism
chemical_list: D004247:DNA