Size distribution and maturation of newly replicated DNA through the S and G2 phases of Physarum polycephalum.

The size distribution of newly made DNA and the dynamics of size maturation of progeny DNA molecules were studied in the synchronous S and G2 phases of Physarum polycephalum. Pulse labeling of DNA and analysis of the products on alkaline sucrose gradients showed that synthesis of primary replication units (which will also be referred to as "Okazaki" fragments) occurred throughout the S period. Pulse and pulse-chase experiments revealed a distinct pattern of size maturation. An apparently linear increase in molecular weight of progeny DNA molecules during the first hour of the S phase occurred at a rate of approximately 4-5 X 10(5) daltons per min at 26 degrees C, corresponding to the joining of 6-8 Okazaki fragments. The resulting 35-45S (1.1-2.2 X 10(7) daltons) DNA molecules may correspond to the Physarum "replicon." The further size increases of the newly made DNA appear to occur in steps, possibly reflecting a clustering of isochronous replicons along the chromatide. These observations are discussed with regard to mechanisms of DNA replication and size maturation.