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Possible function of the c-myc product: promotion of cellular DNA replication

S M Iguchi-Ariga1, T Itani, Y Kiji

  • 1Institute of Medical Science, University of Tokyo, Japan.

The EMBO Journal
|August 1, 1987
PubMed

Insights

The c-myc protein binds to the replication initiation site on the pARS65 plasmid, promoting autonomous DNA replication in human cells. Antibodies against c-myc inhibit this replication process.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • The plasmid pARS65, derived from mouse liver DNA, exhibits autonomous replication in mammalian cells.
  • The c-myc protein is a known oncogene involved in cell proliferation and differentiation.

Purpose of the Study:

  • To investigate the role of the c-myc protein in the replication of the autonomously replicating plasmid pARS65.
  • To determine if c-myc directly interacts with the replication initiation site of pARS65.

Main Methods:

  • Co-transfection of HL-60 cells with pARS65 and anti-c-myc antibody.
  • In vitro replication assays using HL-60 nuclear extracts and pARS65 as a template.
  • Immunobinding assays to detect c-myc binding to pARS65.

Main Results:

  • Replication of pARS65 in HL-60 cells was inhibited by anti-c-myc antibody.
  • In vitro replication of pARS65 was blocked by the addition of anti-c-myc antibody.
  • Specific binding of the c-myc protein to pARS65 was confirmed via immunobinding assays.

Conclusions:

  • The c-myc protein promotes DNA replication by binding to the initiation site on the pARS65 plasmid.
  • Anti-c-myc antibodies can inhibit autonomous DNA replication, suggesting a therapeutic or research application.
  • This study provides evidence for the direct involvement of c-myc in initiating DNA replication from specific sequences.

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