Induction of DNA lesions, chromosomal aberrations, and G2 delay by bromo- and chlorodeoxyuridine

The presence of lesions in DNA of CHO cells, substituted with BrdUrd or CldUrd, has been investigated in a direct way by alkaline sucrose gradient and nucleoid sedimentation analysis and indirectly by screening for induced CAs. The influence of inhibitors of DNA repair (Caff and 3AMB) or DNA synthesis (HU) on the frequencies of such aberrations has been estimated. No randomly located DNA breaks could be detected under neutral conditions, but BrdUrd-substituted DNA was found to contain numerous alkali labile sites. At high concentrations, CldUrd causes G2 delay, similar to the action of known DNA-damaging agents. The extent of delay depends on the pattern of incorporation of the analog, i.e., incorporation for 2 cell cycles causes the longest delay, growth for 12 hr in CldUrd followed by 12 hr in dThd- containing medium gives less delay, and delay is not significant when the cells are incubated in the analog for only 12 hr prior to fixation. Numerous chromatid-type aberrations are present in cells incubated at the highest CldUrd concentration and their induction follows the pattern of induction of G2 delay, indicating the involvement of a common lesion. 3AMB, and HU increase the number of CAs when added 2 hr before fixation. The significance of these results is discussed.