Determination of polycomb group of protein compartmentalization through chromatin fractionation procedure

Federica Marasca, Fabrizia Marullo, Chiara Lanzuolo

Research output: Contribution to journalArticlepeer-review

Abstract

Epigenetic mechanisms modulate and maintain the transcriptional state of the genome acting at various levels on chromatin. Emerging findings suggest that the position in the nuclear space and the cross talk between components of the nuclear architecture play a role in the regulation of epigenetic signatures. We recently described a cross talk between the Polycomb group of proteins (PcG) epigenetic repressors and the nuclear lamina. This interplay is important for the maintenance of transcriptional repression at musclespecific genes and for the correct timing of muscle differentiation. To investigate the synergism between PcG factors and nuclear architecture we improved a chromatin fractionation protocol with the aim to analyze the PcG nuclear compartmentalization. We thus separated PcG proteins in different fractions depending on their solubility. We surprisingly found a consistent amount of PcG proteins in the matrixassociated fraction. In this chapter we describe the chromatin fractionation procedure, a method that can be used to study the nuclear compartmentalization of Polycomb group of proteins and/or PcG targets in murine and Drosophila cells.

Original languageEnglish
Pages (from-to)167-180
Number of pages14
JournalMethods in Molecular Biology
Volume1480
DOIs
Publication statusPublished - 2016
Externally publishedYes

Keywords

  • Chromatin fractionation
  • Lamin
  • Nuclear compartmentalization
  • Polycomb

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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