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Related Experiment Videos

c-Myc-induced extrachromosomal elements carry active chromatin.

Greg Smith1, Cheryl Taylor-Kashton, Len Dushnicky

  • 1Manitoba Institute of Cell Biology, CancerCare Manitoba, the Genomic Center for Cancer Research and Diagnosis, Winnipeg, Manitoba, Canada.

Neoplasia (New York, N.Y.)
|March 28, 2003
PubMed
Summary

Deregulation of c-Myc in murine lymphocytes generates large, self-propagating extrachromosomal elements (EEs) that contain active chromatin and specific genes like DHFR. These findings reveal a novel class of c-Myc-induced genetic structures.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • MycER activation in murine Pre-B lymphocytes induces both chromosomal and extrachromosomal gene amplification.
  • Extrachromosomal elements (EEs) are extrachromosomal DNA molecules that replicate independently of the host chromosome.

Purpose of the Study:

  • To elucidate the size, structure, and functional components of c-Myc-induced extrachromosomal elements (EEs).
  • To characterize the nature of EEs generated by c-Myc deregulation in lymphocytes.

Main Methods:

  • Scanning electron microscopy to determine EE size.
  • Immunohistochemistry and Western blot analyses to assess histone association and chromatin activity.
  • Detection of specific genes (DHFR, CCNC) and DNA replication (BrdU incorporation) within EEs.

Related Experiment Videos

Main Results:

  • c-Myc-induced EEs are significantly larger (10x) than those from control cells and are associated with histone proteins.
  • A proportion of c-Myc-induced EEs exhibit transcriptionally competent/active chromatin.
  • c-Myc-induced EEs can contain the dihydrofolate reductase (DHFR) gene and incorporate bromodeoxyuridine (BrdU), indicating self-propagation.

Conclusions:

  • c-Myc deregulation generates a new class of large, histone-associated EEs in murine lymphocytes.
  • These novel EEs possess characteristics of active chromatin and self-propagation, carrying specific genes like DHFR.
  • The findings suggest a novel mechanism of gene amplification and potential oncogenic activity driven by c-Myc.