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Related Experiment Videos

Barrier function at HMR.

Masaya Oki1, Rohinton T Kamakaka

  • 1Unit on Chromatin and Transcription, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA.

Molecular Cell
|September 3, 2005
PubMed
Summary
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Silencing spread is blocked by a unique tRNA barrier and chromatin modifications. Combined mutations in the barrier and modifiers allow silencing to spread, revealing two overlapping restriction mechanisms.

Area of Science:

  • Molecular Biology
  • Epigenetics
  • Chromatin Biology

Background:

  • The heterochromatin spreading (HMR) domain is a silenced region in yeast.
  • Barrier elements restrict the spread of silenced chromatin.
  • A unique tRNA(THR) gene acts as a right barrier element.

Purpose of the Study:

  • To investigate the mechanisms restricting the spread of silenced chromatin.
  • To elucidate the role of the tRNA(THR) gene barrier and associated chromatin modifications.

Main Methods:

  • Chromatin immunoprecipitation to analyze histone modifications (acetylation, methylation).
  • Nucleosome mapping to identify nucleosome-free regions.
  • Genetic mutations to assess the impact of barrier elements and chromatin modifiers.

Related Experiment Videos

Main Results:

  • The barrier element was linked to a nucleosome-free region, with flanking regions showing histone acetylation.
  • SAGA acetyltransferase was present throughout the region, even after barrier mutation.
  • Single mutations in the barrier or chromatin modifiers did not cause unrestricted silencing spread.
  • Double mutations in the tRNA barrier and specific complexes led to significant Sir protein spread.

Conclusions:

  • Two overlapping mechanisms restrict silenced chromatin spread: a DNA-binding element (tRNA barrier) and specific chromatin-modifying activities.
  • The tRNA barrier creates a nucleosome-free gap, contributing to silencing restriction.
  • Synergy between the barrier element and chromatin modifiers is crucial for robust silencing maintenance.