Role of multi zinc finger proteins in the dynamic change of the nuclear architecture during cell cycle and differentiation

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The multi-zinc finger proteins BORIS and CTCF are unique and conserved factors with a role in transcriptional regulation, the organization of chromatin into distinct domains and imprinting. BORIS is expressed in the testis in cells that do not express CTCF. Abnormal upregulation of BORIS, on the other hand, may be linked to tumorigenesis. Thus, while binding to similar sites in the genome, these proteins could have distinct roles. We have generated inducible BORIS and CTCF knock out mice and are generating GFP- (or biotin)-tagged BORIS and CTCF knock in mice. From the inducible knock out mice cell lines have been isolated, which can be transfected with (mutant) multi-zinc finger proteins and/or DNA contructs with particular binding sites. Using these tools we will perform microscopic (live) imaging analysis (group Galjart), affinity purifications of biotin-tagged proteins (groups Galjart and Renakwitz) and structural analysis, like DNA loop formation (group Renkawitz) on the different types of mice, tissues and cells. This proposal aims at understanding the dynamic behaviour of both multi-zinc finger proteins during the cell cycle and the relevance of this behaviour and of these proteins for the maintenance of chromatin structure.