Enhanced p53 activity and accumulation in response to DNA damage upon DNA transfection.
In response to DNA damage the wild-type tumor suppressor protein p53 accumulates in the nucleus of rodent and primate cells. To investigate the minimal requirement for this reaction the cellular DNA was restricted by two alternative ways: (i) by calicheamicin gamma 1, an enediyne, which causes direct, sequence-specific DNA damage, as shown by fluorimetric analysis of DNA unwinding and by poly(ADP-ribose) polymerase activation. The dose-dependent DNA damage correlated with the nuclear p53 accumulation. In addition, restriction was generated (ii) by the intracellular introduction of the restriction enzyme PvuII, which generates blunt-ended DNA breaks, applying a mild hypotonic shock (pellet method). Previous transfection of linear or circular, single- or ds, DNA, followed by mitomycin C-treatment, lead to a dramatic increase in nuclear p53 accumulation and p53 activity according to electrophoretic mobility shift analysis. The nature of transfected DNA was irrelevant for enhanced accumulation. The data suggest, that the cellular p53 response to DNA damage is sensitized by uptake of exogenous DNA.
D000617:Aminoglycosides; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000187:drug effects / Q000235:genetics*; D004249:DNA Damage / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism; D053281:Enediynes; D016158:Genes, p53; D016685:Mitomycin / Q000494:pharmacology; D008969:Molecular Sequence Data; D011323:Primates; D012377:Rodentia; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D000903:Antibiotics, Antineoplastic; D004268:DNA-Binding Proteins; D053281:Enediynes; D016159:Tumor Suppressor Protein p53; C055955:calicheamicin gamma(1)I; D016685:Mitomycin; D004247:DNA