The reconstitution of higher-order DNA structure after X-irradiation of mammalian cells.

X-ray-induced DNA repair in mouse leukemia (L1210) cells was studied by alkaline elution, which measures the amount of DNA strand breakage, coupled with nucleoid sedimentation, which measures DNA compactness. Two phases of X-ray repair were detected. An initial phase was rapid (t1/2 less than 10 min). During this phase most strand breaks were rejoined and some compaction occurred. After a lag of 1-2 hours, a second phase occurred which exhibited very little or no additional ligation but further compaction of the nucleoid DNA. Both the DNA strand rejoining and initial nucleoid compaction of the first phase were inhibited by 3-ABA2 but not by novobiocin, and the second phase was inhibited by novobiocin but not by 3-ABA. The two phases of reconstitution of nucleoid compactness following X-irradiation are thus distinguishable by their time of occurrence and by their sensitivity to inhibitors of DNA-related enzymes. A coordinated process of ligation followed by compaction may be intrinsic to DNA repair following X-irradiation.