Compositional and functional analysis of ribosomal RNA gene chromatin in distinct transcriptional states (A05)

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We developed a new strategy allowing the in vivo analysis and purification of RNA polymerase (Pol) I transcribed episomal chromatin. Episomal chromatin will be analyzed using established and complementary ex vivo and in vivo approaches. We propose to pursue two main research goals:(i) to analyse structure function relationships within the RNA polymerase (Pol) I preinitiation complex (PIC) and to determine the molecular requirements for its assembly(ii) to define the molecular nature of the open Pol I transcribed chromatin state, its contribution to Pol I transcription, and the mechanisms required for its establishment.We expect high synergy in particular with project AP1 in a comparative functional and structural analysis of Pol I PICs obtained by in vitro reconstitution or after in vivo assembly in yeast. We will further continue our collaboration with project A1 in the reconstitution of Pol I transcription from native chromatin templates in vitro. We suppose that this work will contribute to our understanding of the intimate interrelationship between chromatin structure and the process of nuclear transcription in eukaryotes.