title: Chromatin of the Barr body: histone and non-histone proteins associated with or excluded from the inactive X chromosome.
abstract: The Barr body has long been recognized as the cytological manifestation of the inactive X chromosome (Xi) in interphase nuclei. Despite being known for over 50 years, relatively few components of the Barr body have been identified. In this study, we have screened over 30 histone variants, modified histones and non-histone proteins for their association with or exclusion from the Barr body. We demonstrate that, similar to the histone variant macroH2A, heterochromatin protein-1 (HP1), histone H1 and the high mobility group protein HMG-I/Y are elevated at the territory of the Xi in interphase in human cell lines, but only when the Xi chromatin is heteropycnotic, implicating each as a component of the Barr body. Surprisingly, however, virtually all other candidate proteins involved in establishing heterochromatin and gene silencing are notably absent from the Barr body despite being localized generally elsewhere throughout the nucleus, indicating that the Barr body represents a discrete subnuclear compartment that is not freely accessible to most chromatin proteins. A similar dichotomous pattern of association or exclusion describes the spatial relationship of a number of specific histone methylation patterns in relation to the Barr body. Notably, though, several methylated forms of histone H3 that are deficient in Xi chromatin generally are present at a region near the macrosatellite repeat DXZ4, as are the chromatin proteins CTCF and SAP30, indicating a distinctive chromatin state in this region of the Xi. Taken together, our data imply that the Xi adopts a distinct chromatin configuration in interphase nuclei and are consistent with a mechanism by which HP1, through histone H3 lysine-9 methylation, recognizes and assists in maintaining heterochromatin and gene silencing at the human Xi.
authors.affiliation: Institute for Genome Sciences and Policy, Duke University, Box 3382, Durham, NC 27710, USA.
authors.forename: Brian P
authors.initials: BP
authors.lastname: Chadwick
authors.forename: Huntington F
authors.initials: HF
authors.lastname: Willard
mesh_terms: D000076246:CCCTC-Binding Factor; D002467:Cell Nucleus; D002843:Chromatin / Q000235:genetics; D000090266:Chromobox Protein Homolog 5; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D041321:Chromosomes, Human, X / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000378:metabolism; D004303:Dosage Compensation, Genetic; D020868:Gene Silencing; D006609:High Mobility Group Proteins / Q000378:metabolism*; D006655:Histone Deacetylases / Q000378:metabolism; D006657:Histones / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007399:Interphase; D018895:Microsatellite Repeats; D009707:Nucleosomes; D012097:Repressor Proteins / Q000378:metabolism; D012728:Sex Chromatin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012729:Sex Chromosome Aberrations
chemical_list: D000076246:CCCTC-Binding Factor; C000619697:CTCF protein, human; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D006657:Histones; D009707:Nucleosomes; D012097:Repressor Proteins; C113294:SAP30 protein, human; C120756:macroH2A histone; D000090266:Chromobox Protein Homolog 5; D006655:Histone Deacetylases