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

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 a...

Full description

Bibliographic Details
Main Authors: Dorte Bohla, Martin Herold, Imke Panzer, Melanie K Buxa, Tamer Ali, Jeroen Demmers, Marcus Krüger, Maren Scharfe, Michael Jarek, Marek Bartkuhn, Rainer Renkawitz
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC4172637?pdf=render
id doaj-98b2b417259442eca46af26921b9687d
record_format Article
spelling doaj-98b2b417259442eca46af26921b9687d2020-11-25T02:15:27ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0199e10776510.1371/journal.pone.0107765A functional insulator screen identifies NURF and dREAM components to be required for enhancer-blocking.Dorte BohlaMartin HeroldImke PanzerMelanie K BuxaTamer AliJeroen DemmersMarcus KrügerMaren ScharfeMichael JarekMarek BartkuhnRainer RenkawitzChromatin 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.http://europepmc.org/articles/PMC4172637?pdf=render
collection DOAJ
language English
format Article
sources DOAJ
author Dorte Bohla
Martin Herold
Imke Panzer
Melanie K Buxa
Tamer Ali
Jeroen Demmers
Marcus Krüger
Maren Scharfe
Michael Jarek
Marek Bartkuhn
Rainer Renkawitz
spellingShingle Dorte Bohla
Martin Herold
Imke Panzer
Melanie K Buxa
Tamer Ali
Jeroen Demmers
Marcus Krüger
Maren Scharfe
Michael Jarek
Marek Bartkuhn
Rainer Renkawitz
A functional insulator screen identifies NURF and dREAM components to be required for enhancer-blocking.
PLoS ONE
author_facet Dorte Bohla
Martin Herold
Imke Panzer
Melanie K Buxa
Tamer Ali
Jeroen Demmers
Marcus Krüger
Maren Scharfe
Michael Jarek
Marek Bartkuhn
Rainer Renkawitz
author_sort Dorte Bohla
title A functional insulator screen identifies NURF and dREAM components to be required for enhancer-blocking.
title_short A functional insulator screen identifies NURF and dREAM components to be required for enhancer-blocking.
title_full A functional insulator screen identifies NURF and dREAM components to be required for enhancer-blocking.
title_fullStr A functional insulator screen identifies NURF and dREAM components to be required for enhancer-blocking.
title_full_unstemmed A functional insulator screen identifies NURF and dREAM components to be required for enhancer-blocking.
title_sort functional insulator screen identifies nurf and dream components to be required for enhancer-blocking.
publisher Public Library of Science (PLoS)
series PLoS ONE
issn 1932-6203
publishDate 2014-01-01
description 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.
url http://europepmc.org/articles/PMC4172637?pdf=render
work_keys_str_mv AT dortebohla afunctionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT martinherold afunctionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT imkepanzer afunctionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT melaniekbuxa afunctionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT tamerali afunctionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT jeroendemmers afunctionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT marcuskruger afunctionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT marenscharfe afunctionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT michaeljarek afunctionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT marekbartkuhn afunctionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT rainerrenkawitz afunctionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT dortebohla functionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT martinherold functionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT imkepanzer functionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT melaniekbuxa functionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT tamerali functionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT jeroendemmers functionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT marcuskruger functionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT marenscharfe functionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT michaeljarek functionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT marekbartkuhn functionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
AT rainerrenkawitz functionalinsulatorscreenidentifiesnurfanddreamcomponentstoberequiredforenhancerblocking
_version_ 1724896185871761408