Key mediators of the enhancer blocker function of Drosophila CTCF

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Functional eukaryotic gene units including all of their regulatory elements are insulated from each other to allow the action of regulatory elements on the appropriate gene. We and others have analyzed the DNA binding factor CTCF, which is the only protein known to mediate a directional blocking activity of enhancer elements in vertebrates. Despite many efforts from several laboratories, a mechanism mediating enhancer blocking has not been determined. In contrast, in Drosophila other factors have been descr ibed, mediating chromatin insulation and enhancer blocking. In this application, for the first time, we have the possibility to compare the properties of the unpublished Drosophila orthologue of CTCF with the function of vertebrate CTCF. We will use the advantages of both systems to identify CTCF domains and nucleosomal modifications involved in enhancer blocking in order to unravel the molecular mechanism of enhancer blocking.