Formation of nascent DNA molecules during inhibition of replicon initiation in mammalian cells.
X-irradiation of mammalian cells with moderate doses (100-1000 rads) inhibits the initiation of DNA replicons. This inhibition is observed as depressed amounts of radioactivity at low molecular weights when the DNA from the cells is analysed by velocity sedimentation in alkaline sucrose gradients at 30 min after irradiation. There is no detectable effect on chain elongation and joining of those molecules that do initiate replication; this is indicated by the presence of the same amounts of radioactivity in nascent DNA molecules of high molecular weights from control and irradiated cells. The labeling of DNA molecules that initiated replication before irradiation continues unhindered for more than 60 min after irradiation, which is observed as peaks of radioactivity at high S values in alkaline sucrose gradients from irradiated cells. These data indicate that DNA replication in mammalian cells proceeds by continuous joining of nascent molecules that initiate almost simultaneously at origins at various distances from one another. Some of the interorigin distances are much greater than others, implying that large replicons make up a significant component of mammalian DNA.
D002460:Cell Line; D002499:Centrifugation, Density Gradient; D004260:DNA Repair; D004261:DNA Replication / Q000528:radiation effects*; D004307:Dose-Response Relationship, Radiation; D006367:HeLa Cells / Q000378:metabolism / Q000528:radiation effects; D007739:L Cells / Q000378:metabolism / Q000528:radiation effects; D008970:Molecular Weight; D011830:Radiation Effects