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 Torre, Patrick Onck, Hendrik Dietz () and Cees Dekker ()
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Philip Ketterer: Technische Universität München, Am Coulombwall 4a
Adithya N. Ananth: Delft University of Technology
Diederik S. Laman Trip: Delft University of Technology
Ankur Mishra: University of Groningen, Nijenborgh 4
Eva Bertosin: Technische Universität München, Am Coulombwall 4a
Mahipal Ganji: Delft University of Technology
Jaco Torre: Delft University of Technology
Patrick Onck: University of Groningen, Nijenborgh 4
Hendrik Dietz: Technische Universität München, Am Coulombwall 4a
Cees Dekker: Delft University of Technology

Nature Communications, 2018, vol. 9, issue 1, 1-8

Abstract: 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.

Date: 2018
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DOI: 10.1038/s41467-018-03313-w

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