Epigenetic Complex Assembly in Space and Time (EpiCAST)

• Applicant: Professor Dr. Marc Timmers
• Subject Area: General Genetics and Functional Genome Biology ; Biochemistry ; Cell Biology
• Term: from 2019 to 2024
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 431324432

The collaborative action of distinct transcriptional regulators controls transcription by RNA polymerase II (Pol II), which underpins the identity and function of all eukaryotic cells. Regulated transcription depends on chromatin and transcription regulatory complexes recruited by gene-specific transcription factors to direct the assembly of Pol II preinitiation complexes. Interestingly, many chromatin and transcription complexes function as large multi-protein assemblies, which often share subunits. Thus, an elaborate decision-making process must exist, which governs the assembly of chromatin and transcription regulatory complexes. But very little is known concerning the cellular rules governing their biogenesis. Technological advances sparked a world-wide interest into the regulated biogenesis of multi-protein complexes as an exciting novel area of research. To tackle this fundamental problem in biology, the German and French partners will expand their long-standing collaborations by combining cutting-edge technologies to charting co-translation-driven and chaperone-assisted assembly pathways of multi-subunit complexes in space and time. Together, they will determine the assembly intermediates, involved chaperones, and cytoplasmic and nuclear dynamics of (sub-)complexes. The Partners will focus on crucial general transcription factor (TFIID) and histone-modifying complexes (SAGA, ATAC, SET1/MLL), all linked to important diseases. The EpiCAST collaboration builds upon common projects, publications and participations in European networks.In EpiCAST the German and the French partner will investigate the biogenesis pathways of these key regulatory complexes with the following aims:1. Investigate co-translation-driven assembly of complexes and their assembly intermediates.2. Determine the chaperones required for assembly of transcription and chromatin complexes3. Understand transcription complex assembly dynamics in living cellsThe biosynthesis of these multi-subunit complexes will be systematically scrutinized in a combination of quantitative proteomics, RNA immunoprecipitation, cell engineering, single-molecule fluorescence in situ hybridization, super-resolution live-cell imaging, and synthetic gene arrays. We will obtain quantitative data to build kinetic frameworks for the assembly pathways of complexes, together covering more than 60 subunits. We will challenge these frameworks in perturbation experiments by examining disease-related mutants in TFIID subunits, by analysing truncations of WD40-domain proteins and by testing small-molecule inhibitors of WDR5 central to eight chromatin regulators. This project will provide the cellular rules guiding the biogenesis of key transcription and chromatin regulatory complexes, which will be applicable to large complexes of various functions. Thus, we foresee that EpiCAST will have a major impact on the biomedical field and will lead to new paradigms in cell biology.

• DFG Programme: Research Grants
• International Connection: France
• Cooperation Partner: Professor Laszlo Tora, Ph.D.