Initiation and termination of vaccinia virus DNA replication

The initiation and termination sites of replication of vaccinia DNA has been studied by determining the radioactivity in restriction fragments of the pulse-labeled newly synthesized molecules. The results indicate a bimodal distribution of radioactivity in the molecules completed during a pulse shorter than the completion time of synthesis, implying termination at both ends. The symmetry of replication was studied by hybridization of an intermediate in replication (34 S) to the isolated strands of virion DNA followed by restriction analysis. The frequency of hybridization of the fragments to the light or heavy strand shows a unimodal distribution of radioactivity indicating asymmetric replication. Together, these observations suggest that initiation and termination sites are located at both ends of the chromosome. Changes observed in the conformation of parental DNA at the time of replication, such as high degree of single-strandness, are compatible with strand displacement during synthesis.