Recovery of a DNA-protein complex in cultured mammalian cells from damage caused by 4-nitroquinoline 1-oxide.
Treatment of FM3A cells with 4-nitroquinoline 1-oxide caused a decrease in the sedimentation velocity of a DNA-protein complex, but did not cause a dissociation of the complex, as revealed by neutral sucrose gradient centrifugation. Microscopic autoradiography of the complex spread on a Millipore filter, demonstrated that treatment of the cells with 4-nitroquinoline 1-oxide, or of the complex with Pronase E, gave rise to a relaxed mass of DNA fibers, in contrast to a compact mass of DNA from control cells. The damage to the DNA-protein compelx was repaired completely by incubation of the cells in a medium without 4-nitroquinoline 1-oxide. The following metabolic inhibitors had no effect on the repair of the complex: inhibitors of nucleic acid synthesis, alpha-amanitine, cordycepin, 2-mercapto-1-(beta-4-pyridethyl)benzimidazol, 1-beta-D-arabinofuranosylcytosine, 5-fluorodeoxyuridine, and hydroxyurea; inhibitors of protein synthesis, cycloheximide and puromycin; an inhibitor of the dark repair process in a variety of biological systems, caffeine; inhibitors of the microtubular and microfilament system, Colcemid and cytochalasin B, respectively; and inhibitors of energy metabolism, 2,4-dinitrophenol, KCN, iodoacetic acid, ouabain, and an atmosphere of nitrogen. Acriflavine and actinomycin D, which are known to intercalate into DNA, caused a decrease in the sedimentation velocity of the DNA-protein complex; therefore, the effects of these agents on the recovery process remained unsolved. The repair process of the complex was, however, demonstrated to be temperature dependent. The process was inhibited at 10 degrees, retarded at 28 degrees, but accelerated at 40 degrees as compared with the rate at 37 degrees.
D015112:4-Nitroquinoline-1-oxide / Q000494:pharmacology*; D000167:Acriflavine / Q000494:pharmacology; D000546:Amanitins / Q000494:pharmacology; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D002110:Caffeine / Q000494:pharmacology; D002478:Cells, Cultured / Q000187:drug effects*; D003078:Colchicine / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D004247:DNA; D004260:DNA Repair / Q000187:drug effects; D003839:Deoxyadenosines / Q000494:pharmacology; D004140:Dinitrophenols / Q000494:pharmacology; D066298:In Vitro Techniques; D007461:Iodoacetates / Q000494:pharmacology; D009600:Nitroquinolines / Q000494:pharmacology*; D009698:Nucleoproteins; D010042:Ouabain / Q000494:pharmacology; D011691:Puromycin / Q000494:pharmacology; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D013696:Temperature
D000546:Amanitins; D000970:Antineoplastic Agents; D003839:Deoxyadenosines; D004140:Dinitrophenols; D007461:Iodoacetates; D009600:Nitroquinolines; D009698:Nucleoproteins; D013438:Sulfhydryl Compounds; D000167:Acriflavine; D003571:Cytochalasin B; D002110:Caffeine; D011691:Puromycin; D015112:4-Nitroquinoline-1-oxide; D010042:Ouabain; D004247:DNA; D003513:Cycloheximide; D003078:Colchicine