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Related Experiment Videos

Do transcriptional enhancers also augment DNA replication?

D T O'Connor1, S Subramani

  • 1Veterans Administration Medical Center, San Diego, CA.

Nucleic Acids Research
|December 9, 1988
PubMed
Summary
This summary is machine-generated.

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The simian virus 40 (SV40) enhancer can boost DNA replication, but only when positioned very close to the replication origin. This proximity, not typical enhancer activity, appears crucial for stimulating DNA replication.

Area of Science:

  • Molecular Biology
  • Genetics
  • Virology

Background:

  • Enhancers are DNA sequences that increase gene transcription.
  • Previous evidence suggested enhancers might influence DNA replication.
  • Hormone-dependent cancer growth and viral replication stimulation by enhancers hinted at a replication role.

Purpose of the Study:

  • To investigate if DNA sequences with enhancer properties can also stimulate DNA replication.
  • To determine if enhancer-like sequences share properties with DNA replication-stimulating elements.
  • To test the hypothesis that enhancers directly affect DNA replication.

Main Methods:

  • Recombinant plasmids containing the simian virus 40 (SV40) core enhancer and SV40 minimal origin of replication (ori) were constructed.

Related Experiment Videos

  • Plasmids were introduced into T antigen-producing COS cells for replication assays.
  • DNA replication was quantified by measuring progeny DNA accumulation or the ratio of replicated to unreplicated DNA.

    • Main Results:

    • The SV40 enhancer increased DNA replication 1.5-10 fold when ligated adjacent to the replication origin.
    • Replication stimulation was dependent on proximity to the origin, time, and partially on orientation.
    • A heterologous glucocorticoid enhancer showed no effect on replication.
    • Enhancer copy number did not influence the replication effect.

    Conclusions:

    • The SV40 enhancer's cis-effect on DNA replication requires close proximity to the replication origin.
    • Specific homologous sequences within the enhancer, near the origin, are likely responsible for replication stimulation.
    • The mechanism appears distinct from typical enhancer-mediated transcriptional activation.