PT  - JOURNAL ARTICLE
AU  - Qi Shen
AU  - Taoran Tian
AU  - Qiancheng Xiong
AU  - Patrick D. Ellis Fisher
AU  - Yong Xiong
AU  - Thomas J. Melia
AU  - C. Patrick Lusk
AU  - Chenxiang Lin
TI  - A DNA-origami NanoTrap for studying the diffusion barriers formed by Phe-Gly-rich nucleoporins
AID  - 10.1101/2021.02.07.430177
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.02.07.430177
4099  - http://biorxiv.org/content/early/2021/02/08/2021.02.07.430177.short
4100  - http://biorxiv.org/content/early/2021/02/08/2021.02.07.430177.full
AB  - DNA nanotechnology provides a versatile and powerful tool to dissect the structure-function relationship of biomolecular machines like the nuclear pore complex (NPC), an enormous protein assembly that controls molecular traffic between the nucleus and cytoplasm. To understand how the intrinsically disordered, Phe-Gly-rich nucleoporins (FG-nups) within the NPC’s central transport channel impede the diffusion of macromolecules, we built a DNA-origami NanoTrap. The NanoTrap comprises precisely arranged FG-nups in an NPC-like channel, which sits on a baseplate that captures macromolecules that pass through the FG network. Using this biomimetic construct, we determined that the FG-motif type, grafting density and spatial arrangement are critical determinants of an effective diffusion barrier. Further, we observe that diffusion barriers formed with cohesive FG-interactions dominate in mixed-FG-nup scenarios. Our DNA-origami platform thus sheds light on how NPCs sieve inert macromolecules and will provide a valuable tool for studying nuclear transport.Competing Interest StatementThe authors have declared no competing interest.