title: Determination of Polycomb Group of Protein Compartmentalization Through Chromatin Fractionation Procedure.
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 muscle-specific 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 matrix-associated 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.
authors.affiliation: Istituto Nazionale di Genetica Molecolare "Romeo and Enrica Invernizzi", Via Francesco Sforza 35, 20122, Milan, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Marasca
authors.affiliation: Institute of Cell Biology and Neurobiology, IRCCS Santa Lucia Foundation, via del Fosso di Fiorano 64, Rome, 00143, Italy.
authors.forename: Fabrizia
authors.initials: F
authors.lastname: Marullo
authors.affiliation: Istituto Nazionale di Genetica Molecolare "Romeo and Enrica Invernizzi", Via Francesco Sforza 35, 20122, Milan, Italy. chiara.lanzuolo@cnr.it.|Institute of Cell Biology and Neurobiology, IRCCS Santa Lucia Foundation, via del Fosso di Fiorano 64, Rome, 00143, Italy. chiara.lanzuolo@cnr.it.
authors.forename: Chiara
authors.initials: C
authors.lastname: Lanzuolo
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002843:Chromatin / Q000235:genetics / Q000302:isolation & purification*; D004330:Drosophila; D044127:Epigenesis, Genetic; D051379:Mice; D008967:Molecular Biology / Q000379:methods*; D063146:Polycomb-Group Proteins / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D002843:Chromatin; D063146:Polycomb-Group Proteins
keywords: Chromatin fractionation; Lamin; Nuclear compartmentalization; Polycomb