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Related Experiment Videos

Sequence elements in cis influence heterochromatic silencing in trans.

Brian T Sage1, John L Jones, Amy L Holmes

  • 1Department of Biological Sciences, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA.

Molecular and Cellular Biology
|December 17, 2004
PubMed
Summary

The brown(Dominant) allele silences nearby genes through heterochromatin association. Researchers identified a specific DNA sequence that enhances this silencing effect, offering insights into gene regulation.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Epigenetics

Background:

  • The brown(Dominant) (bw(D)) allele in Drosophila melanogaster features a large heterochromatin insertion.
  • This insertion causes aberrant heterochromatin association and silencing of the wild-type allele in heterozygotes.
  • Transgenes near the bw(D) locus can also be silenced, a phenomenon termed trans inactivation, which varies with transgene regulatory sequences and proximity to the insertion.

Purpose of the Study:

  • To investigate endogenous cis-acting sequences that influence a reporter gene's susceptibility to trans inactivation.
  • To identify specific DNA elements that modulate the silencing effect of the bw(D) allele.

Main Methods:

  • Generation of new Drosophila lines by inducing flanking deletions in parental lines with P-element transgenes.

Related Experiment Videos

  • Analysis of mini-white reporter gene expression and trans inactivation levels in these new lines.
  • Correlation of deleted regions with reporter gene silencing to identify cis-acting elements.

    • Main Results:

    • Different endogenous sequences flanking the reporter gene resulted in varied susceptibility to bw(D)-mediated silencing.
    • A 301-bp sequence was identified as an enhancer of trans inactivation.
    • This enhancer region contains upstream sequences of divergently transcribed genes and a potential BEAF binding motif.

    Conclusions:

    • Cis-acting DNA elements significantly influence the susceptibility of reporter genes to trans inactivation.
    • A novel 301-bp sequence enhances trans inactivation, potentially through interactions involving chromatin boundary proteins like BEAF.
    • These findings contribute to understanding the mechanisms of heterochromatin-mediated gene silencing and epigenetic regulation.