Abstract

Using a combination of Bal31 deletion mutagenesis and site-directed mutagenesis, we analyzed the sequence requirements for the DNA replication of the human neurotropic polyomavirus JC. In addition to defining the late side boundary of the viral core origin we demonstrated that the viral enhancer stimulates replication in vivo. Three regions within the viral enhancer increased the rate of replication, with sequences directly adjacent to the late side of the core origin exhibiting the strongest effect. These sequences interact with various cellular proteins, among them NF-I. Point mutations within the NF-I site abolished the stimulation of DNA replication concomitant with a strong reduction in NF-I binding. By contrast, point mutations which did not interfere with NF-I binding did not influence the rate of replication in vivo. Stimulation of JCV DNA replication could only be observed in vivo, but not in vitro, indicating a role of NF-I in determining chromatin structure.

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