MNase-Seq Analysis for Identifying Stress-Altered Nucleosome Occupancy in Plants

Methods Mol Biol. 2024:2832:33-46. doi: 10.1007/978-1-0716-3973-3_2.

Abstract

Nucleosome occupancy plays an important role in chromatin compaction, affecting biological processes by hampering the binding of cis-acting elements such as transcription factors, RNA polymerase machinery, and coregulatory. Accessible regions allow for cis-acting elements to bind DNA and regulate transcription. Here, we detail our protocol to profile nucleosome occupancy and chromatin structure dynamics under drought stress at the genome-wide scale using micrococcal nuclease (MNase) digestion. Combining variable MNase concentration treatments and high-throughput sequencing, we investigate the changes in the overall chromatin state using bread wheat samples from an exemplary drought experiment.

Keywords: Chromatin structural variation; MNase digestion; MNase-seq; Nucleosome occupancy.

MeSH terms

  • Chromatin / genetics
  • Chromatin / metabolism
  • Droughts
  • Gene Expression Regulation, Plant
  • High-Throughput Nucleotide Sequencing* / methods
  • Micrococcal Nuclease* / metabolism
  • Nucleosomes* / genetics
  • Nucleosomes* / metabolism
  • Plants / genetics
  • Plants / metabolism
  • Stress, Physiological* / genetics
  • Triticum* / genetics
  • Triticum* / metabolism

Substances

  • Nucleosomes
  • Micrococcal Nuclease
  • Chromatin