DNA replication of human polyomavirus JC is stimulated by NF-I in vivo.
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.
Sock
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Institut für Biochemie, Universität Würzburg, Germany.
Wegner
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Grummt
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F
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000502:physiology; D001483:Base Sequence; D001665:Binding Sites; D022762:CCAAT-Enhancer-Binding Proteins; D002460:Cell Line; D002522:Chlorocebus aethiops; D004252:DNA Mutational Analysis; D004261:DNA Replication; D004279:DNA, Viral / Q000235:genetics; D004268:DNA-Binding Proteins / Q000502:physiology*; D004742:Enhancer Elements, Genetic; D015967:Gene Expression Regulation, Viral; D006801:Humans; D066298:In Vitro Techniques; D007577:JC Virus / Q000235:genetics / Q000254:growth & development*; D008969:Molecular Sequence Data; D051839:NFI Transcription Factors; D009687:Nuclear Proteins; D014157:Transcription Factors / Q000502:physiology*; D014779:Virus Replication; D051840:Y-Box-Binding Protein 1
D000952:Antigens, Polyomavirus Transforming; D022762:CCAAT-Enhancer-Binding Proteins; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D051839:NFI Transcription Factors; D009687:Nuclear Proteins; D014157:Transcription Factors; D051840:Y-Box-Binding Protein 1; C070856:YBX1 protein, human