title: Analysis of histone posttranslational modifications from nucleolus-associated chromatin by mass spectrometry.
abstract: Chromatin is unevenly distributed within the eukaryote nucleus and it contributes to the formation of morphologically and functionally distinct substructures, called chromatin domains and nuclear bodies. Here we describe an approach to assess specific chromatin features, the histone posttranslational modifications (PTMs), of the largest nuclear sub-compartment, the nucleolus. In this chapter, methods for the isolation of nucleolus-associated chromatin from native or formaldehyde-fixed cells and the effect of experimental procedures on the outcome of mass spectrometry analysis of histone PTMs are compared.
authors.affiliation: Biochemistry Center Regensburg, University of Regensburg, Regensburg, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Dillinger
authors.forename: Ana Villar
authors.initials: AV
authors.lastname: Garea
authors.forename: Rainer
authors.initials: R
authors.lastname: Deutzmann
authors.forename: Attila
authors.initials: A
authors.lastname: Németh
mesh_terms: D000595:Amino Acid Sequence; D002466:Cell Nucleolus / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005557:Formaldehyde / Q000737:chemistry; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational
chemical_list: D002843:Chromatin; D006657:Histones; D005557:Formaldehyde