Presence of UV-endonuclease sensitive sites in daughter DNA of UV-irradiated mammalian cells. [3H-thymidine tracer]

Asynchronous Chinese hamster cells were irradiated with 10 Jm/sup -2/ uv radiation and 0.25 to 4 hours later pulse-labeled with (/sup 3/H)thymidine. Cells synchronized by shaking off mitotic and G/sub 1/ cells were irradiated in either the G/sub 1/-phase or S-phase of the cell cycle and pulse-labeled with (/sup 3/H)thymidine in the S-phase. After a 12 to 14 hour chase in unlabeled medium, the DNA was extracted, incubated with Micrococcus luteus uv-endonuclease and sedimented in alkaline sucrose. The number of endonuclease sensitive sites decreased as the time between uv irradiation and pulse-labeling of daughter DNA increased. Further, there were significantly less endonuclease sensitive sites in the daughter DNA from cells irradiated in the G/sub 1/-phase than in the S-phase. These data indicate that very few, if any, dimers are transferred from parental DNA to daughter DNA and that the dimers detected in daughter DNA may be due to the irradiation of replicating daughter DNA before labeling