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A novel ex vivo approach to study the role of chromatin in the regulation of transcription


The DNA of the eukaryotic cell is accommodated by the structure of chromatin. To administrate the genetic information the cell employs multiple mechanisms regulating protein-DNA transactions in the context of chromatin. Research of the past two decades has demonstrated that chromatin undergoes dynamic changes depending on its functional state: The major chromatin components, the histones, are subject to covalent modifications by specific enzymatic activities, furthermore multi-protein complexes have been de scribed that structurally alter chromatin in ATP-dependent processes. A detailed molecular analysis eventually requires isolation of pure, natural chromatin closely reflecting the in vivo situation. We have established a novel technique to purify single-copy genes from chromosomal loci in the yeast Saccharomyces cerevisiae to near homogeneity, conserving the chromatin structure of distinct transcriptional states.

Future research pursues two lines of investigations:
(i) Determination of structure and composition of natural chromatin in different functional states by electron microscopy and mass spectrometry
(ii) Description of the functional interplay between chromatin remodelling and transcription using isolated natural chromosomal domains and purified protein factor in vitro

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