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Related Experiment Videos

Relationship between retroviral DNA integration and gene expression.

J B Weidhaas1, E L Angelichio, S Fenner

  • 1Department of Molecular Biology and Microbiology, Tufts University, Boston, Massachusetts 02111, USA.

Journal of Virology
|August 23, 2000
PubMed
Summary

Retroviral DNA integration is not enhanced by transcriptional activity. Studies show that transcriptionally active DNA may actually decrease retroviral DNA integration into the host genome.

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Area of Science:

  • Molecular Biology
  • Virology
  • Genetics

Background:

  • Retroviral DNA integration into the host genome is essential for viral replication but the specificity of integration sites is poorly understood.
  • Transcriptional activity of the host genome is hypothesized to influence retroviral integration, but evidence remains controversial.

Purpose of the Study:

  • To investigate the effect of transcriptional activity on retroviral DNA integration in vivo.
  • To determine if actively transcribed DNA is a preferred target for retroviral integration.

Main Methods:

  • Created quail cell clones with a minigene cassette regulated by the papilloma virus E2 protein.
  • Derived transcriptionally active and silent subclones expressing wild-type or mutant E2 proteins.
  • Infected subclones with avian leukosis virus and used PCR to analyze viral DNA integration patterns.

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Main Results:

  • Integration into the gene cassette decreased in transcriptionally active subclones expressing wild-type E2 protein.
  • Integration also decreased in subclones expressing mutant E2 protein, specifically within the DNA binding region.
  • Contrary to expectations, transcriptional activity did not enhance, but rather decreased, retroviral integration.

Conclusions:

  • Transcriptionally active DNA is not a preferred target for retroviral integration.
  • Transcriptional activity may be inversely correlated with retroviral DNA integration efficiency.
  • These findings challenge existing hypotheses regarding retroviral integration specificity.