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Related Experiment Videos

RNA polymerase III interferes with Ty3 integration

C M Connolly1, S B Sandmeyer

  • 1Department of Microbiology and Molecular Genetics, College of Medicine, University of California, Irvine 92697-4025, USA.

FEBS Letters
|April 1, 1997
PubMed
Summary

Budding yeast Ty3 retrotransposon integration requires specific promoter elements. RNA polymerase III (pol III) inhibits Ty3 integration, suggesting competition for the transcription initiation site.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Yeast Biology

Background:

  • Ty3 is a gypsy-like retrotransposon in budding yeast.
  • Ty3 retrotransposon integration occurs at the transcription initiation site of RNA polymerase III (pol III) transcribed genes.
  • In vitro studies show Ty3 integration requires intact promoter elements and pol III transcription factors (TFIIIB and TFIIIC).

Purpose of the Study:

  • To investigate the effect of pol III on Ty3 retrotransposon integration in vitro.
  • To elucidate the mechanism of Ty3 integration concerning pol III activity.

Main Methods:

  • In vitro Ty3 integration assays were performed.
  • Increasing concentrations of a pol III-containing fraction were added to the reactions.
  • The impact of pol III on integration efficiency was measured.

Main Results:

  • The addition of a pol III-containing fraction inhibited Ty3 in vitro integration.
  • The inhibitory effect was dose-dependent, with increasing pol III amounts reducing integration.

Conclusions:

  • Budding yeast pol III negatively regulates Ty3 retrotransposon integration.
  • Ty3 integration complex and pol III likely compete for binding to similar features of the transcription initiation site.
  • This competition mechanism explains the observed inhibition of Ty3 integration by pol III.

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