A Method to Identify Nucleolus-Associated Chromatin Domains (NADs)

Methods Mol Biol. 2018:1675:99-109. doi: 10.1007/978-1-4939-7318-7_7.

Abstract

The nuclear context needs to be taken into consideration to better understand the mechanisms shaping the epigenome and its organization, and therefore its impact on gene expression. For example, in Arabidopsis, heterochromatin is preferentially localized at the nuclear and the nucleolar periphery. Although chromatin domains associating with the nuclear periphery remain to be identified in plant cells, Nucleolus Associated chromatin Domains (NADs) can be identified thanks to a protocol allowing the isolation of pure nucleoli. We describe here the protocol enabling the identification of NADs in Arabidopsis. Providing the transfer of a nucleolus marker as described here in other crop species, this protocol is broadly applicable.

Keywords: Chromatin; FACS; NADs; Nucleolus; Nucleolus-associated chromatin domains; Nucleus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics
  • Cell Nucleolus / genetics*
  • Cell Nucleus / genetics
  • Chromatin / chemistry*
  • Chromatin / genetics
  • Computational Biology / methods
  • DNA, Plant / genetics
  • Genome, Plant
  • Sequence Analysis, DNA / methods*
  • Transcription, Genetic

Substances

  • Chromatin
  • DNA, Plant