A designer FG-Nup that reconstitutes the selective transport barrier of the nuclear pore complex

Alessio Fragasso, Hendrik W. Vries, John Andersson, Eli O. Sluis, Erik Giessen, Andreas Dahlin, Patrick R. Onck () and Cees Dekker ()
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Alessio Fragasso: Delft University of Technology
Hendrik W. Vries: University of Groningen
John Andersson: Chalmers University of Technology
Eli O. Sluis: Delft University of Technology
Erik Giessen: University of Groningen
Andreas Dahlin: Chalmers University of Technology
Patrick R. Onck: University of Groningen
Cees Dekker: Delft University of Technology

Nature Communications, 2021, vol. 12, issue 1, 1-15

Abstract: Abstract Nuclear Pore Complexes (NPCs) regulate bidirectional transport between the nucleus and the cytoplasm. Intrinsically disordered FG-Nups line the NPC lumen and form a selective barrier, where transport of most proteins is inhibited whereas specific transporter proteins freely pass. The mechanism underlying selective transport through the NPC is still debated. Here, we reconstitute the selective behaviour of the NPC bottom-up by introducing a rationally designed artificial FG-Nup that mimics natural Nups. Using QCM-D, we measure selective binding of the artificial FG-Nup brushes to the transport receptor Kap95 over cytosolic proteins such as BSA. Solid-state nanopores with the artificial FG-Nups lining their inner walls support fast translocation of Kap95 while blocking BSA, thus demonstrating selectivity. Coarse-grained molecular dynamics simulations highlight the formation of a selective meshwork with densities comparable to native NPCs. Our findings show that simple design rules can recapitulate the selective behaviour of native FG-Nups and demonstrate that no specific spacer sequence nor a spatial segregation of different FG-motif types are needed to create selective NPCs.

Date: 2021
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DOI: 10.1038/s41467-021-22293-y

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