A functional insulator screen identifies NURF and dREAM components to be required for enhancer-blocking.

2.50
Hdl Handle:
http://hdl.handle.net/10033/337962
Title:
A functional insulator screen identifies NURF and dREAM components to be required for enhancer-blocking.
Authors:
Bohla, Dorte; Herold, Martin; Panzer, Imke; Buxa, Melanie K; Ali, Tamer; Demmers, Jeroen; Krüger, Marcus; Scharfe, Maren; Jarek, Michael; Bartkuhn, Marek; Renkawitz, Rainer
Abstract:
Chromatin insulators of higher eukaryotes functionally divide the genome into active and inactive domains. Furthermore, insulators regulate enhancer/promoter communication, which is evident from the Drosophila bithorax locus in which a multitude of regulatory elements control segment specific gene activity. Centrosomal protein 190 (CP190) is targeted to insulators by CTCF or other insulator DNA-binding factors. Chromatin analyses revealed that insulators are characterized by open and nucleosome depleted regions. Here, we wanted to identify chromatin modification and remodelling factors required for an enhancer blocking function. We used the well-studied Fab-8 insulator of the bithorax locus to apply a genome-wide RNAi screen for factors that contribute to the enhancer blocking function of CTCF and CP190. Among 78 genes required for optimal Fab-8 mediated enhancer blocking, all four components of the NURF complex as well as several subunits of the dREAM complex were most evident. Mass spectrometric analyses of CTCF or CP190 bound proteins as well as immune precipitation confirmed NURF and dREAM binding. Both co-localise with most CP190 binding sites in the genome and chromatin immune precipitation showed that CP190 recruits NURF and dREAM. Nucleosome occupancy and histone H3 binding analyses revealed that CP190 mediated NURF binding results in nucleosomal depletion at CP190 binding sites. Thus, we conclude that CP190 binding to CTCF or to other DNA binding insulator factors mediates recruitment of NURF and dREAM. Furthermore, the enhancer blocking function of insulators is associated with nucleosomal depletion and requires NURF and dREAM.
Affiliation:
Helmholtz Centre for ifection research, Innhoffenstr. 7, D38124 Braunschweig, Germany.
Citation:
A functional insulator screen identifies NURF and dREAM components to be required for enhancer-blocking. 2014, 9 (9):e107765 PLoS ONE
Journal:
PloS one
Issue Date:
2014
URI:
http://hdl.handle.net/10033/337962
DOI:
10.1371/journal.pone.0107765
PubMed ID:
25247414
Type:
Article
Language:
en
ISSN:
1932-6203
Appears in Collections:
publications of the research group genomeanalytics (GMAK)

Full metadata record

DC FieldValue Language
dc.contributor.authorBohla, Dorteen
dc.contributor.authorHerold, Martinen
dc.contributor.authorPanzer, Imkeen
dc.contributor.authorBuxa, Melanie Ken
dc.contributor.authorAli, Tameren
dc.contributor.authorDemmers, Jeroenen
dc.contributor.authorKrüger, Marcusen
dc.contributor.authorScharfe, Marenen
dc.contributor.authorJarek, Michaelen
dc.contributor.authorBartkuhn, Mareken
dc.contributor.authorRenkawitz, Raineren
dc.date.accessioned2015-01-09T09:18:43Z-
dc.date.available2015-01-09T09:18:43Z-
dc.date.issued2014-
dc.identifier.citationA functional insulator screen identifies NURF and dREAM components to be required for enhancer-blocking. 2014, 9 (9):e107765 PLoS ONEen
dc.identifier.issn1932-6203-
dc.identifier.pmid25247414-
dc.identifier.doi10.1371/journal.pone.0107765-
dc.identifier.urihttp://hdl.handle.net/10033/337962-
dc.description.abstractChromatin insulators of higher eukaryotes functionally divide the genome into active and inactive domains. Furthermore, insulators regulate enhancer/promoter communication, which is evident from the Drosophila bithorax locus in which a multitude of regulatory elements control segment specific gene activity. Centrosomal protein 190 (CP190) is targeted to insulators by CTCF or other insulator DNA-binding factors. Chromatin analyses revealed that insulators are characterized by open and nucleosome depleted regions. Here, we wanted to identify chromatin modification and remodelling factors required for an enhancer blocking function. We used the well-studied Fab-8 insulator of the bithorax locus to apply a genome-wide RNAi screen for factors that contribute to the enhancer blocking function of CTCF and CP190. Among 78 genes required for optimal Fab-8 mediated enhancer blocking, all four components of the NURF complex as well as several subunits of the dREAM complex were most evident. Mass spectrometric analyses of CTCF or CP190 bound proteins as well as immune precipitation confirmed NURF and dREAM binding. Both co-localise with most CP190 binding sites in the genome and chromatin immune precipitation showed that CP190 recruits NURF and dREAM. Nucleosome occupancy and histone H3 binding analyses revealed that CP190 mediated NURF binding results in nucleosomal depletion at CP190 binding sites. Thus, we conclude that CP190 binding to CTCF or to other DNA binding insulator factors mediates recruitment of NURF and dREAM. Furthermore, the enhancer blocking function of insulators is associated with nucleosomal depletion and requires NURF and dREAM.en
dc.language.isoenen
dc.titleA functional insulator screen identifies NURF and dREAM components to be required for enhancer-blocking.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Centre for ifection research, Innhoffenstr. 7, D38124 Braunschweig, Germany.en
dc.identifier.journalPloS oneen

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