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Related Experiment Videos

Programmed factor binding to simian virus 40 GC-box replication and transcription control sequences.

R L Buchanan1, J D Gralla

  • 1Department of Chemistry and Biochemistry, University of California, Los Angeles 90024.

Journal of Virology
|January 1, 1990
PubMed
Summary

Viral DNA replication is linked to factor binding at simian virus 40 GC-boxes. This regulatory region controls replication and factor binding during infection.

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Area of Science:

  • Molecular Biology
  • Virology
  • Genetics

Background:

  • The simian virus 40 (SV40) regulatory region contains repetitive GC-box DNA elements crucial for viral gene expression and replication.
  • Understanding the precise timing and order of factor binding to these elements in vivo is essential for elucidating viral regulatory mechanisms.

Purpose of the Study:

  • To investigate the temporal program of factor binding to SV40 GC-box elements during the late phase of infection.
  • To determine the relationship between SV40 DNA replication and factor binding to the GC-box region.
  • To explore the role of the GC-box region in controlling viral DNA replication.

Main Methods:

  • Nuclear footprinting techniques were employed to analyze factor binding to SV40 GC-box DNA in vivo.
  • Experiments involved the use of DNA replication inhibitors (aphidicolin) and the inactivation of viral replication factors (SV40 T antigen).

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  • Measurements of viral DNA accumulation were performed in modified viruses lacking specific GC-box elements.

    • Main Results:

    • Nuclear footprinting revealed an ordered and directional temporal program of factor binding to SV40 GC-box elements during the late phase of infection.
    • Disruption of viral DNA replication, either by aphidicolin or by inactivating SV40 T antigen, interrupted this factor binding program.
    • Analysis of DNA accumulation indicated a dual role for the GC-box region in regulating viral DNA replication.

    Conclusions:

    • Factor binding to the SV40 GC-box DNA is temporally regulated and linked to viral DNA replication.
    • The GC-box region functions as a multifunctional regulatory element involved in both factor binding and replication control.
    • These findings highlight the intricate interplay between DNA replication and sequence-specific factor binding in viral regulatory pathways.