DNA origami scaffold for studying intrinsically disordered proteins of the nuclear pore complex

Philip Ketterer, Adithya N Ananth, Diederik S Laman Trip, Ankur Mishra, Eva Bertosin, Mahipal Ganji, Jaco van der Torre, Patrick Onck, Hendrik Dietz, Cees Dekker

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Abstract

The nuclear pore complex (NPC) is the gatekeeper for nuclear transport in eukaryotic cells. A key component of the NPC is the central shaft lined with intrinsically disordered proteins (IDPs) known as FG-Nups, which control the selective molecular traffic. Here, we present an approach to realize artificial NPC mimics that allows controlling the type and copy number of FG-Nups. We constructed 34 nm-wide 3D DNA origami rings and attached different numbers of NSP1, a model yeast FG-Nup, or NSP1-S, a hydrophilic mutant. Using (cryo) electron microscopy, we find that NSP1 forms denser cohesive networks inside the ring compared to NSP1-S. Consistent with this, the measured ionic conductance is lower for NSP1 than for NSP1-S. Molecular dynamics simulations reveal spatially varying protein densities and conductances in good agreement with the experiments. Our technique provides an experimental platform for deciphering the collective behavior of IDPs with full control of their type and position.

Original languageEnglish
Article number902
Number of pages8
JournalNature Communications
Volume9
Issue number1
DOIs
Publication statusPublished - 2-Mar-2018

Keywords

  • Journal Article
  • SOLID-STATE NANOPORES
  • SELECTIVE TRANSPORT
  • ACCESS RESISTANCE
  • NANOSCALE SHAPES
  • FOLDING DNA
  • PERMEABILITY
  • NANOSTRUCTURES
  • ARCHITECTURE
  • Gatekeepers
  • MECHANISM

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