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Related Experiment Videos

Enhancer-activated plasmid transcription complexes contain constrained supercoiling.

P J Bonilla1, S O Freytag, L C Lutter

  • 1Molecular Biology Research Program, Henry Ford Hospital, Detroit, MI 48202.

Nucleic Acids Research
|July 25, 1991
PubMed
Summary
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Transcriptionally active genes may have unconstrained supercoiling. This study found enhancer-activated transcription complexes contain constrained supercoils, suggesting a nucleosome-like organization, but unconstrained supercoils cannot be ruled out.

Area of Science:

  • Molecular Biology
  • Genetics
  • Chromatin Structure

Background:

  • Transcriptionally active chromatin is hypothesized to possess unconstrained supercoiling.
  • Recent findings suggest this topological state is characteristic of highly transcribed genes.

Purpose of the Study:

  • To investigate whether enhancer-activated transcription complexes contain unconstrained supercoils.
  • To analyze the DNA topology within transcription complexes of enhancer-activated plasmids.

Main Methods:

  • Transfection of COS7 cells with an enhancer-activated plasmid.
  • Direct determination of DNA topology using agarose gel electrophoresis.
  • Assessment of supercoil behavior after topoisomerase I treatment.

Main Results:

Related Experiment Videos

  • Enhancer-activated plasmid transcription complexes exhibit supercoiling.
  • These supercoils persist even after treatment with topoisomerase I.
  • The observed supercoiling levels are indicative of a nucleosome-like organization within the complexes.

Conclusions:

  • Transcribing complexes contain constrained supercoils, similar to nucleosomes.
  • The presence of additional unconstrained supercoils in the cellular transcription complex cannot be excluded.