A functional insulator screen identifies NURF and dREAM components to be required for enhancer-blocking
Dorte Bohla1, Martin Herold1, Imke Panzer1
1Institut für Genetik, Justus-Liebig-Universität, Giessen, Giessen, Germany.
Plos One
|September 24, 2014
Summary
Chromatin insulators use proteins like CP190 to recruit NURF and dREAM complexes, which are essential for enhancer blocking and creating open chromatin regions.
Area of Science:
- * Molecular Biology
- * Epigenetics
- * Genomics
Background:
- * Chromatin insulators divide the genome into active and inactive domains.
- * Insulators regulate enhancer/promoter communication, controlling gene activity.
- * Centrosomal protein 190 (CP190) is recruited to insulators by DNA-binding factors like CTCF.
Purpose of the Study:
- * Identify chromatin modification and remodeling factors crucial for insulator enhancer blocking function.
- * Investigate the role of NURF and dREAM complexes in insulator activity.
- * Understand the mechanism of CP190-mediated recruitment of these complexes.
Main Methods:
- * Genome-wide RNAi screen using the Fab-8 insulator in Drosophila.
- * Mass spectrometry and co-immunoprecipitation to identify protein interactions.
- * Chromatin immunoprecipitation and nucleosome occupancy assays.
Main Results:
- * Identified 78 genes essential for enhancer blocking, including NURF and dREAM complex components.
- * Confirmed NURF and dREAM bind to CP190 and co-localize with its genomic sites.
- * Demonstrated CP190 recruits NURF and dREAM, leading to nucleosome depletion at binding sites.
Conclusions:
- * CP190 recruits NURF and dREAM complexes to insulators.
- * Insulator enhancer blocking function is linked to nucleosome depletion.
- * NURF and dREAM are required for the enhancer blocking activity of insulators.
