Establishment of a monoclonal antibody recognizing cyclobutane-type thymine dimers in DNA: a comparative study with 64M-1 antibody specific for (6-4)photoproducts.
We obtained a monoclonal antibody (TDM-1) binding to 313-nm UV-irradiated DNA in the presence of acetophenone. The binding of TDM-1 to 254-nm UV-irradiated DNA was not reduced with the subsequent irradiation of 313-nm UV. Furthermore, the treatment of UV-irradiated DNA with photolyase from E. coli and visible light exposure reduced both the antibody binding and the amount of thymine dimers in the DNA. A competitive inhibition assay revealed that the binding of TDM-1 to UV-irradiated DNA was inhibited with photolyase, but not with 64M-1 antibody specific for (6-4)photoproducts. These results suggest that TDM-1 antibody recognizes cyclobutane-type thymine dimers in DNA. Using TDM-1 and 64M-1 antibodies, we differentially measured each type of damage in DNA extracted from UV-irradiated mammalian cells. Repair experiments confirm that thymine dimers are excised from UV-irradiated cellular DNA more slowly than (6-4)photoproducts, and that the excision rates of thymine dimers and (6-4)photoproducts are lower in mouse NIH3T3 cells than in human cells.
Mizuno
T
T
Division of Radiation Biology, Faculty of Pharmaceutical Sciences, Kanazawa University, Japan.
Matsunaga
T
T
Ihara
M
M
Nikaido
O
O
D000098:Acetophenones / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D001667:Binding, Competitive; D002459:Cell Fusion; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004249:DNA Damage; D004260:DNA Repair; D004255:Deoxyribodipyrimidine Photo-Lyase / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D066298:In Vitro Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D009841:Oligonucleotides / Q000378:metabolism; D011740:Pyrimidine Dimers / Q000276:immunology*; D014466:Ultraviolet Rays
D000098:Acetophenones; D000911:Antibodies, Monoclonal; D009841:Oligonucleotides; D011740:Pyrimidine Dimers; D004255:Deoxyribodipyrimidine Photo-Lyase; C038699:acetophenone