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Kinetics of adenovirus dna replication II. Initiation of adenovirus DNA replication

1980; Elsevier BV; Volume: 105; Issue: 2 Linguagem: Inglês

10.1016/0042-6822(80)90037-9

1096-0341

John W. Bodner, George D. Pearson,

Monoclonal and Polyclonal Antibodies Research

We have developed three methods to estimate the initiation of adenovirus DNA replication: (a) direct measurement during density-shift experiments, (b) calculation using the saturation kinetics of viral DNA accumulation, and (c) calculation using the steady-state kinetics of viral DNA replication. The values for the rate of initiation measured by each method agree quantitatively and confirm the postulate that initiation is the rate-limiting step in adenovirus replication. Newly replicated viral DNA molecules are preferentially initiated. This preference is slight early in infection, but increases markedly over the course of infection. We interpret these results to mean that adenovirus is partitioned into several pools, and at least one pool contains molecules destined for replication. The average rate of initiation in this pool is 0.0042 ± 0.0007 initiation/min/end and is constant throughout infection. The maximum size of the replication pool is 50,000 ± 7000 molecules/cell. Steady-state calculations indicate that displacement synthesis is faster than complementary synthesis. The differential between these rates of synthesis leads to premature displacement of nascent strands in type I/II replicating molecules, a process we call premature displacement synthesis. We propose that this process provides the raw material for generating defective adenovirus DNA molecules.