Integrating single-molecule and single-cell measurements to characterise the mechanism of the Nuclear Pore Complex

The Nuclear Pore Complex (NPC) is a truly remarkable molecular machine. It is the major regulatory structure controlling access to the cell nucleus, capable of sorting molecular components and partitioning them against a thermodynamic gradient, all with high efficiency and selectivity. It is essential in correctly partitioning molecules such as transcription factorsfor normal cellular function, and defects in transport have recently been shown to be a fundamental pathway in neurodegenerative diseases, viral infection and various cancers. The molecular mechanisms of transport, assembly and regulation remain elusive due to the technical challenge of visualising such dynamic structures. In this talk, I will detail our efforts to develop a powerful integrated imaging approach, combining single-molecule, super-resolution imaging and quantitative live-cell imaging to characterise the basis of NPC selectivity, the properties of the permeability barrier, and how transport is regulated at the cellular level.