Analysis of chromatin structure in plant cells

Methods Mol Biol. 2012:833:201-23. doi: 10.1007/978-1-61779-477-3_13.

Abstract

A vast body of evidence in the literature indicates that nucleosomes can act as barriers to transcriptional initiation. The nucleosome at the promoter inhibits association of transcription factors disallowing active transcription of the gene. We have found a nucleosome on tobacco pathogenesis-related gene-1a (PR-1a) core promoter and mapped its boundaries and extension to find its span. The nucleosome covers the TATA box and Inr region of the core promoter and gets disassembled upon induction. Prior to its removal, modifications (i.e., acetylation and methylation of histones) occur at the nucleosome, proving a role of epigenetic modifications in transcriptional regulation. We summarize here various methodologies to analyze promoter chromatin structure in plants using the PR-1a core promoter as an example.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / immunology
  • Arabidopsis / cytology
  • Arabidopsis / metabolism
  • Base Sequence
  • Chromatin / chemistry*
  • Chromatin Immunoprecipitation
  • DNA Primers / metabolism
  • DNA, Plant / isolation & purification
  • Electrophoresis, Polyacrylamide Gel
  • Histones / metabolism
  • Micrococcal Nuclease / metabolism
  • Molecular Biology / methods*
  • Molecular Sequence Data
  • Nucleosomes / metabolism
  • Plant Cells / metabolism*
  • Polymerase Chain Reaction
  • Protein Binding
  • Protein Processing, Post-Translational
  • Time Factors

Substances

  • Antibodies
  • Chromatin
  • DNA Primers
  • DNA, Plant
  • Histones
  • Nucleosomes
  • Micrococcal Nuclease