Specific modulation of p53 binding to consensus sequence within supercoiled DNA by monoclonal antibodies.
Monoclonal antibodies (MAbs) were used to investigate the binding of insect cell-expressed, wild-type human p53 protein to the consensus sequence (p53CON) in a 474-bp DNA fragment and to supercoiled (sc) DNAs with and without p53CON. Supershifting of p53-DNA complexes by MAbs in agarose gels was applied to studies of activation of p53 for sequence-specific binding within scDNA. C-terminal specific antibody Bp53-10.1 activated the sequence-specific binding of p53 to p53CON within pPGM1 scDNA but did not influence binding of p53 to pBluescript scDNA (not containing p53CON). Incubation of p53 with DO-1 prior to addition of Bp53-10.1 prevented activation of p53 and induced dissociation of a portion of pPGM1 scDNA from the sequence-specific immune complex; no such dissociation was observed if pPGM1 scDNA was replaced by the 474-bp p53CON-containing DNA fragment.
Brázda
V
V
Institute of Biophysics, Academy of Sciences of the Czech Republic, Královopolská 135, Brno, 612 65, Czech Republic.
Paleĉek
J
J
Pospísilová
S
S
Vojtêsek
B
B
Paleĉek
E
E
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001483:Base Sequence; D001665:Binding Sites; D001666:Binding Sites, Antibody; D002460:Cell Line; D016384:Consensus Sequence; D004278:DNA, Superhelical / Q000737:chemistry* / Q000378:metabolism*; D004279:DNA, Viral / Q000737:chemistry / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007313:Insecta; D007700:Kinetics; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D004278:DNA, Superhelical; D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53