The role of RNA-binding proteins in spatially compartmentalized control of transcription

• Applicant: Professor Lennart Hilbert, Ph.D.
• Subject Area: Biophysics ; Cell Biology
• Term: since 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 469120288

It is essential for organisms to control which genes are expressed by their constituent cells. In eukaryotes, most genes are transcribed by RNA polymerase II, which is controlled by recruitment into a paused state, and subsequent pause-release towards transcription of genes. The recruitment and pause-release occur in the context of macromolecular clusters that are supposedly formed by phase separation mechanisms. Recent work found that the different steps of transcriptional control are localized into distinct spatial compartments, and involve distinct sets of RNA-binding proteins. In this project, we will directly assess the association of several candidate RNA-binding proteins with the different compartments of clusters as a mechanism of their contribution to the different steps of transcriptional control. Specifically, we will apply molecular biology and microscopy techniques to mouse embryonic stem cell lines with protein degradation tags and use in vitro assessments of biochemical properties that could explain phase separation behavior of recombinantly expressed candidate proteins. Our experiments will comprehensively assess RNA-binding proteins as molecular effectors of previously proposed phase separation mechanisms for the spatial compartmentalization of transcriptional control. An integrated biophysical and biochemical understanding of the role of RNA-binding proteins in transcriptional control would help to clarify the molecular mechanisms by which eukaryotic cells ensure robust and precise gene expression.

• DFG Programme: Research Grants
• International Connection: China
• Partner Organisation: National Natural Science Foundation of China
• Cooperation Partner: Professorin Xiaohua Shen, Ph.D.