Establishment and characterization of a monoclonal antibody recognizing the Dewar isomers of (6-4)photoproducts.

We established a monoclonal antibody (DEM-1) that recognizes UV-induced DNA damage other than cyclobutane pyrimidine dimers or (6-4)photoproducts. The binding of DEM-1 antibody to 254 nm UV-irradiated DNA increased with subsequent exposure to UV wavelengths longer than 310 nm, whereas that of the 64M-2 antibody specific for the (6-4)photoproduct decreased with this treatment. Furthermore, the increase in DEM-1 binding was inhibited by the presence of the 64M-2 antibody during the exposure. We concluded that the DEM-1 antibody specifically recognized the Dewar photoproduct, which is the isomeric form of the (6-4)photoproduct. The DEM-1 antibody, however, also bound to DNA irradiated with high fluences of 254 nm UV, suggesting that 254 nm UV could induce Dewar photoproducts without subsequent exposure to longer wavelengths of UV. Furthermore, an action spectral study demonstrated that 254 nm was the most efficient wavelength for Dewar photoproduct induction in the region from 254 to 365 nm, as well as cyclobutane dimers and (6-4)photoproducts, although the action spectrum values in the UV-B region were significantly higher compared with those for cyclobutane dimer and (6-4)photoproduct induction.