Base sequence specificity of a monoclonal antibody binding to (6-4)photoproducts.
The DNA base sequence specificity of the 64M-1 monoclonal antibody, which recognizes ultraviolet (UV)-induced (6-4)photoproducts, was characterized. The 64M-1 antibody strongly bound to UV-poly(dU) as well as to UV-poly(dT), and weakly to UV-poly(dC), UV-poly(me5dC) and UV-poly(rU). A competitive inhibition assay using UV-oligo(dT)8, UV-oligo(dTdC)4, UV-oligo(dC)8, UV-PvuI linker (GCGATCGC) and UV-PvuII linker (GCAGCTGC) indicated that the main (6-4)photoproducts detected by the 64M-1 antibody in UV-irradiated DNA are TT(6-4)photoproducts and TC(6-4)photoproducts. Comparison between dTpdT(6-4)photoproduct and dTpdC(6-4)photoproduct showed that the affinity of the 64M-1 antibody for dTpdT(6-4)photoproduct was about 5 times higher than that for dTpdC(6-4)photoproduct. The antibody also binds to isolated TT(6-4)photoproducts.
Matsunaga
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Division of Radiation Biology, Faculty of Pharmaceutical Sciences, Kanazawa University, Japan.
Mori
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Nikaido
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D004247:DNA / Q000276:immunology* / Q000528:radiation effects; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D009841:Oligonucleotides / Q000528:radiation effects*; D011119:Polynucleotides / Q000528:radiation effects*; D011740:Pyrimidine Dimers / Q000276:immunology*; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured; D014466:Ultraviolet Rays
D000911:Antibodies, Monoclonal; D000939:Epitopes; D009841:Oligonucleotides; D011119:Polynucleotides; D011740:Pyrimidine Dimers; D004247:DNA