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Cellular protein interactions with herpes simplex virus type 1 oriS

C E Dabrowski1, P J Carmillo, P A Schaffer

  • 1Division of Molecular Genetics, Dana-Farber Cancer Institute, Boston, Massachusetts.

Molecular and Cellular Biology
|April 1, 1994
PubMed
Summary

Cellular proteins bind to the herpes simplex virus type 1 origin of replication (oriS), influencing viral DNA synthesis. Origin factor I (OF-I) plays a crucial role in this process.

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Origin binding protein-containing protein-DNA complex formation at herpes simplex virus type 1 oriS: role in oriS-dependent DNA replication.

Journal of virology·2001

Area of Science:

  • Virology
  • Molecular Biology
  • Genetics

Background:

  • Herpes simplex virus type 1 (HSV-1) DNA replication initiates at the oriS region.
  • oriS contains binding sites for the viral origin-binding protein (OBP).
  • The role of cellular proteins in HSV-1 replication initiation was previously unclear.

Purpose of the Study:

  • To characterize interactions between oriS DNA sequences and cellular proteins in uninfected cells.
  • To identify cellular factors involved in HSV-1 DNA replication initiation.
  • To elucidate the function of these cellular factors at oriS.

Main Methods:

  • Gel shift assays were used to detect protein-DNA complex formation.
  • Site-directed mutagenesis was employed to create oriS plasmid variants.

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  • Transient replication assays were performed to assess oriS function.

    • Main Results:

    • A predominant cellular protein-DNA complex, termed origin factor I (OF-I), was identified binding to site I of oriS.
    • OF-I binding sites partially overlap with OBP binding sites and are homologous to CCAAT DNA-binding protein sites.
    • Mutations in the OF-I binding site reduced oriS replication efficiency, indicating its functional importance.

    Conclusions:

    • Cellular proteins, specifically OF-I, participate in the initiation of HSV-1 DNA synthesis.
    • OF-I binding to oriS is essential for efficient viral DNA replication.
    • These findings suggest a collaborative mechanism between viral and cellular factors in HSV-1 replication.