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Related Experiment Videos

Evidence for long-range oncogene activation by hepadnavirus insertion

G Fourel1, J Couturier, Y Wei

  • 1Unité de Recombinaison et Expression Génétique (INSERM U163), Institut Pasteur, Paris, France.

The EMBO Journal
|June 1, 1994
PubMed
Summary

Woodchuck hepatitis virus DNA causes liver tumors by activating the N-myc2 oncogene. Viral integrations near the N-myc2 gene, even at a distance, promote its overexpression and contribute to woodchuck tumorigenesis.

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Area of Science:

  • Hepatology
  • Oncology
  • Molecular Biology

Background:

  • Woodchuck hepatitis virus (WHV), a hepadnavirus, is known to cause insertional mutagenesis and liver tumors in woodchucks.
  • WHV frequently activates myc genes, particularly the N-myc2 oncogene, in over 50% of woodchuck liver tumors.

Purpose of the Study:

  • To investigate the mechanism of N-myc2 overexpression in woodchuck liver tumors where genetic alterations are not apparent.
  • To elucidate the role of WHV in N-myc2 activation and woodchuck tumorigenesis.

Main Methods:

  • Cloning and analysis of single viral integration sites in woodchuck tumors.
  • Fluorescent in situ hybridization (FISH) to localize N-myc2 and the identified integration locus (win).
  • Pulse-field gel electrophoresis to determine the physical distance between N-myc2 and the win locus.

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Main Results:

  • Viral integration sites were clustered within a 20 kb region at a single locus (win) in four tumors with wild-type myc alleles.
  • FISH localized N-myc2 and the win locus to the same region on the woodchuck X chromosome, with an estimated 150-180 kb intervening distance.
  • Viral integrations in the win locus correlated with abundant N-myc2 transcripts in additional tumors.

Conclusions:

  • WHV can activate the N-myc2 oncogene through long-range enhancer activity, even when integrated at a distance.
  • This long-range activation mechanism is implicated in woodchuck liver cell transformation and tumorigenesis.