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Related Experiment Videos

Analyzing chromatin structure and transcription factor binding in yeast

P D Gregory1, S Barbaric, W Hörz

  • 1Institut für Physiologische Chemie, Universität München, Germany.

Methods (San Diego, Calif.)
|September 19, 1998
PubMed
Summary

Investigate chromatin structure and DNA function using two in vivo methods: DNase I for isolated nuclei and dimethyl sulfate footprinting for whole cells. These techniques correlate transactivator binding with chromatin organization changes.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Chromatin, initially viewed as a structural component for DNA compaction, is crucial for understanding DNA function within the cell.
  • Investigating chromatin organization provides insights into gene regulation and DNA-protein interactions.

Purpose of the Study:

  • To present two fundamental in vivo procedures for studying chromatin structure and function.
  • To enable the correlation of transactivator binding with specific alterations in chromatin organization.

Main Methods:

  • DNase I-based method for analyzing chromatin structure in isolated nuclei.
  • Dimethyl sulfate (DMS) footprinting on whole cells to map factor binding to cis-regulatory elements in vivo.

Main Results:

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  • The described methods successfully resolve chromatin structure and identify factor binding sites.
  • Application to the yeast PHO8 gene and other yeast loci demonstrates reliability and effectiveness.
  • Demonstrated ability to link transactivator binding events with changes in chromatin organization.

Conclusions:

  • The presented techniques offer valuable tools for dissecting the dynamic interplay between DNA-binding factors and chromatin structure.
  • These methods advance the understanding of gene regulation at specific genomic loci.