Recognition of cisplatin-damaged DNA by p53 protein: critical role of the p53 C-terminal domain.
It was shown previously that the p53 protein can recognize DNA modified with antitumor agent cisplatin (cisPt-DNA). Here, we studied p53 binding to the cisPt-DNA using p53 deletion mutants and via modulation of the p53-DNA binding by changes of the protein redox state. Isolated p53 C-terminal domain (CTD) bound to the cisPt-DNA with a significantly higher affinity than to the unmodified DNA. On the other hand, p53 constructs involving the core domain but lacking the C-terminal DNA binding site (CTDBS) exhibited only small binding preference for the cisPt-DNA. Oxidation of cysteine residues within the CD of posttranslationally unmodified full length p53 did not affect its ability to recognize cisPt-DNA. Blocking of the p53 CTDBS by a monoclonal antibody Bp53-10.1 resulted in abolishment of the isolated CTD binding to the cisPt-DNA. Our results demonstrate a crucial role of the basic region of the p53 CTD (aa 363-382) in the cisPt-DNA recognition.
Pivonková
Hana
H
Institute of Biophysics, Academy of Sciences of the Czech Republic, Královopolská 135, 612 65 Brno, Czech Republic.
Brázdová
Marie
M
Kaspárková
Jana
J
Brabec
Viktor
V
Fojta
Miroslav
M
D000911:Antibodies, Monoclonal / Q000276:immunology; D001665:Binding Sites; D002945:Cisplatin / Q000494:pharmacology*; D004247:DNA / Q000378:metabolism*; D004249:DNA Damage / Q000187:drug effects*; D010084:Oxidation-Reduction; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D017384:Sequence Deletion / Q000235:genetics; D013379:Substrate Specificity; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D016159:Tumor Suppressor Protein p53; D004247:DNA; D002945:Cisplatin