My proposal presents a strategy to set up the first minimal system of mRNA export through a biomimetic nuclear pore complex (NPC), which will allow to resolve crucial questions on nuclear export. Separation of genetic material in the cell nucleus allows mRNA to be modified before export to the cytoplasm for translation. When these modifications have been completed, NPCs (that maintain a selective barrier for the transport of macromolecules between the nuclear and cytoplasmic compartments) provide a highly selective channel through the nuclear envelope. Whereas proteins are imported into the nucleus after translation in the cytoplasm, mRNAs are exported after transcription and processing in the nucleus is finished. Unlike protein import, mRNA export proceeds by addition and removal of mRNA-binding proteins until an export-competent messenger ribonucleoprotein particle (mRNP) is formed by attaching the heterodimeric Mex67/Mtr2 transport factor. Crucial questions in this fields are: what is the mechanism by which Mex67/Mtr2 mediates translocation? What are the minimal requirements for transport? What is the Mex67/Mtr2/mRNA stoichiometry within mRNPs? Which conformations do mRNP complexes assume during transport? The molecular complexity of NPCs has defeated classical cellular and molecular approaches addressing such questions. I propose to tackle them using the powerful, innovative setup offered by the biomimetic NPCs, minimal NPCs obtained by coating a solid-state nanopore with NPC transport channel components. This functional artificial pore can deliver powerful information about translocations at a single-molecule level and has already been used successfully to study protein import. I propose, for the first time, to tailor biomimetic nanopores to study mRNA export dynamics to help disentangle mRNPs translocation from their production, maturation, and disassembling steps, allowing key transport questions to be addressed. This minimal pore system will enable the nature of the minimal mRNP export mechanism to be unravelled, and presents unique opportunities for fine-tuned control.

Damage in the DNA inhibits transcription of genes by RNA polymerase II, which copies the genetic information of DNA into RNA. This impediment of RNA polymerase II results in severe cellular dysfunction and accelerated aging. Through a consortium combining unique complementary knowledge and expertise, we can study for the first time the causes and consequences of DNA damage from the perspective of a single molecule to that of a whole organism. Using this approach, we will study what exactly happens to RNA polymerase when encountering DNA damage, and directly link this to the consequences at the cellular and organism level.

For organisms to develop well, cell-fate decisions need to be made correctly, and these in turn are based on an accurate sense of the cell’s environment. Researchers will investigate mechanistic processes which contribute to noise, regarding the spatial structure of the genome, on the one hand, and develop information-theoretical and mathematic frameworks to infer optimal network architectures from experimental data on the other. This project is based on experimental collaborations in a series of developing systems, from flies to viruses.

Our brain is an enormous factory, with proteins and protein complexes executing various processing and maintenance jobs. How they work, and why they sometimes make mistakes, depends on their complex 3D structures and intricate adaptations to specific job descriptions. The researchers will reconstruct important proteins in 3D using advanced electron microscopy and data processing tools tol help to gain insight into the important protein machinery of the brain, and – eventually - to develop new therapeutics for neurological disorders.

In this project, we aim to bring nanoscale neuroscience research closer to the public and involve various groups in society. The scientific goal is to understand how specific chemical structures, called sugars, which are found on the surface of many brain proteins, affect the interaction between nerve cells. This research is important because issues with these proteins can lead to various neurological diseases. The project will not only expand our knowledge but also help the public understand why this research is important.