Project Details
Projekt
Structure and dynamics of the human rDNA transcription machinery
Subject Area
Biochemistry
Structural Biology
Structural Biology
Term
since 2025
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 556771037
In all eukaryotes, the synthesis of the ribosomal RNA precursor is carried out by RNA polymerase (Pol) I. However, regulation of activity, transcription factor composition and molecular mechanisms of this highly complex machinery substantially differ among organisms. We propose to investigate the specific structural and functional traits of the human Pol I transcription system. We ask what the roles of metazoan-specific factors, domains and mobile, flexibly linked elements in the mechanisms of transcription initiation and promoter escape are. Based on a human cell line created in our laboratory, human Pol I and its transcription factors can be purified and are now available for reconstitution and in-depth functional analysis using designed transcription assays and structural analyses. Integrating results from biochemical studies and single-particle cryo-EM will allow a structural description and analysis of structure-function relationships. In addition, the behavior of mobile elements, such as the conserved, flexibly linked tandem winged helix domain of Pol I subunit RPA49, will be studied throughout the initiation and elongation phases of the enzymes’ activity cycle using single-molecule Förster-Resonance-Energy-Transfer (FRET). Taken together, we propose a comprehensive structure-function-dynamics analysis to unravel the mechanisms of human Pol I in unprecedented molecular detail allowing us to compare different transcription systems and the evolution of the ribosomal precursor synthesis machineries among eukaryotes. The results will not only fill a gap in the understanding of an essential process in biology at the molecular level, but also lay the ground for understanding their role in malfunction during human disease.
DFG Programme
Research Grants
