Figure 1.

Expression, sequence specific DNA binding activity, and nuclear localization of murine p53 deletion mutants. (a) Schematic representation of the p53 mutants used. Numbering of murine p53 amino acids is done according to ref. 32. (b) In vitro translated murine p53 mutants were analyzed by SDS/PAGE to confirm the expected apparent molecular weights. The structure of deletion Δ(8–63)/5980 is described in Fig. 3. (c) In vitro translated proteins were tested for sequence specific DNA binding by electrophoretic mobility shift assay with the p53CON oligonucleotide as a probe. Specificity of the binding was confirmed by competition with excess cold p53CON or p53MUT oligonucleotides. (d) 100 ng/μl of each pGDSV7 expression plasmid (31) containing the different p53 deletion mutants, was microinjected into the nuclei of BALB/c (10)1 cells (36). p53 protein expression was analyzed 12 hr later by indirect immunofluorescence using PAb240. Images were obtained with a LSM 450 confocal microscope (Zeiss).