DNA-mediated oxidation of p53.
Transcription factor p53 is the most commonly altered gene in human cancer. As a redox-active protein in direct contact with DNA, p53 can directly sense oxidative stress through DNA-mediated charge transport. Electron hole transport occurs over long distances through the π-stacked bases and leads to the oxidative dissociation of p53. The extent of protein dissociation depends upon the redox potential of the DNA in direct contact with each p53 monomer. The DNA sequence dependence of p53 oxidative dissociation was examined by electrophoretic mobility shift assays using oligonucleotides containing both synthetic and human p53 consensus sequences with an appended photooxidant, anthraquinone. Greater p53 dissociation is observed from sequences containing low-redox potential purine regions, particularly guanine triplets. Using denaturing polyacrylamide gel electrophoresis of irradiated anthraquinone-modified DNA, the DNA damage sites corresponding to sites of preferred electron hole localization were determined. The resulting DNA damage preferentially localizes to guanine doublets and triplets. Oxidative DNA damage is inhibited in the presence of p53, but only at sites in direct contact with p53. From these data, predictions about the sensitivity of human p53-binding sites to oxidative stress as well as possible biological implications have been made. On the basis of our data, the guanine pattern within the purine region of each p53-binding site determines the response of p53 to DNA oxidation, yielding for some sequences the oxidative dissociation of p53 from a distance and thereby providing another potential role for DNA charge transport chemistry within the cell.
Division of Chemistry and Chemical Engineering, California Institute of Technology , Pasadena, California 91125, United States.
Kathryn N
KN
Schaefer
Jacqueline K
JK
Barton
D000880:Anthraquinones / Q000737:chemistry; D001483:Base Sequence; D016384:Consensus Sequence; D004247:DNA / Q000737:chemistry* / Q000528:radiation effects; D004249:DNA Damage; D006801:Humans; D009841:Oligonucleotides / Q000737:chemistry; D016877:Oxidants / Q000737:chemistry; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011401:Promoter Regions, Genetic; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry*
D000880:Anthraquinones; D009841:Oligonucleotides; D016877:Oxidants; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D004247:DNA