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Title: The liquid state of FG-nucleoporins mimics permeability barrier properties of nuclear pore complexes

Abstract

Nuclear pore complexes (NPCs) regulate all cargo traffic across the nuclear envelope. The transport conduit of NPCs is highly enriched in disordered phenylalanine/glycine-rich nucleoporins (FG-Nups), which form a permeability barrier of still elusive and highly debated molecular structure. Here we present a microfluidic device that triggered liquid-to-liquid phase separation of FG-Nups, which yielded droplets that showed typical properties of a liquid state. On the microfluidic chip, droplets were perfused with different transport-competent or -incompetent cargo complexes, and then the permeability barrier properties of the droplets were optically interrogated. We show that the liquid state mimics permeability barrier properties of the physiological nuclear transport pathway in intact NPCs in cells: that is, inert cargoes ranging from small proteins to large capsids were excluded from liquid FG-Nup droplets, but functional import complexes underwent facilitated import into droplets. Collectively, these data provide an experimental model of how NPCs can facilitate fast passage of cargoes across an order of magnitude in cargo size.

Authors:
ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1];  [2]; ORCiD logo [2]; ORCiD logo [1]
  1. Biocentre, Johannes Gutenberg-University Mainz, Mainz, Germany, Institute of Molecular Biology, Mainz, Germany, Structural and Computational Biology Unit and Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany
  2. Center for Integrated Nanotechnologies, Sandia National Laboratories, Albuquerque, NM
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1574083
Resource Type:
Published Article
Journal Name:
Journal of Cell Biology
Additional Journal Information:
Journal Name: Journal of Cell Biology; Journal ID: ISSN 0021-9525
Publisher:
Rockefeller University Press
Country of Publication:
United States
Language:
English

Citation Formats

Celetti, Giorgia, Paci, Giulia, Caria, Joana, VanDelinder, Virginia, Bachand, George, and Lemke, Edward A. The liquid state of FG-nucleoporins mimics permeability barrier properties of nuclear pore complexes. United States: N. p., 2019. Web. doi:10.1083/jcb.201907157.
Celetti, Giorgia, Paci, Giulia, Caria, Joana, VanDelinder, Virginia, Bachand, George, & Lemke, Edward A. The liquid state of FG-nucleoporins mimics permeability barrier properties of nuclear pore complexes. United States. doi:10.1083/jcb.201907157.
Celetti, Giorgia, Paci, Giulia, Caria, Joana, VanDelinder, Virginia, Bachand, George, and Lemke, Edward A. Wed . "The liquid state of FG-nucleoporins mimics permeability barrier properties of nuclear pore complexes". United States. doi:10.1083/jcb.201907157.
@article{osti_1574083,
title = {The liquid state of FG-nucleoporins mimics permeability barrier properties of nuclear pore complexes},
author = {Celetti, Giorgia and Paci, Giulia and Caria, Joana and VanDelinder, Virginia and Bachand, George and Lemke, Edward A.},
abstractNote = {Nuclear pore complexes (NPCs) regulate all cargo traffic across the nuclear envelope. The transport conduit of NPCs is highly enriched in disordered phenylalanine/glycine-rich nucleoporins (FG-Nups), which form a permeability barrier of still elusive and highly debated molecular structure. Here we present a microfluidic device that triggered liquid-to-liquid phase separation of FG-Nups, which yielded droplets that showed typical properties of a liquid state. On the microfluidic chip, droplets were perfused with different transport-competent or -incompetent cargo complexes, and then the permeability barrier properties of the droplets were optically interrogated. We show that the liquid state mimics permeability barrier properties of the physiological nuclear transport pathway in intact NPCs in cells: that is, inert cargoes ranging from small proteins to large capsids were excluded from liquid FG-Nup droplets, but functional import complexes underwent facilitated import into droplets. Collectively, these data provide an experimental model of how NPCs can facilitate fast passage of cargoes across an order of magnitude in cargo size.},
doi = {10.1083/jcb.201907157},
journal = {Journal of Cell Biology},
number = ,
volume = ,
place = {United States},
year = {2019},
month = {11}
}

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Works referenced in this record:

Integrative structure and functional anatomy of a nuclear pore complex
journal, March 2018

  • Kim, Seung Joong; Fernandez-Martinez, Javier; Nudelman, Ilona
  • Nature, Vol. 555, Issue 7697
  • DOI: 10.1038/nature26003

Cell type‐specific nuclear pores: a case in point for context‐dependent stoichiometry of molecular machines
journal, January 2013

  • Ori, Alessandro; Banterle, Niccolò; Iskar, Murat
  • Molecular Systems Biology, Vol. 9, Issue 1
  • DOI: 10.1038/msb.2013.4

Structure and gating of the nuclear pore complex
journal, June 2015

  • Eibauer, Matthias; Pellanda, Mauro; Turgay, Yagmur
  • Nature Communications, Vol. 6, Issue 1
  • DOI: 10.1038/ncomms8532

Formation and Maturation of Phase-Separated Liquid Droplets by RNA-Binding Proteins
journal, October 2015


Molecular architecture of the inner ring scaffold of the human nuclear pore complex
journal, April 2016


DNA origami scaffold for studying intrinsically disordered proteins of the nuclear pore complex
journal, March 2018

  • Ketterer, Philip; Ananth, Adithya N.; Laman Trip, Diederik S.
  • Nature Communications, Vol. 9, Issue 1
  • DOI: 10.1038/s41467-018-03313-w

Architecture of the symmetric core of the nuclear pore
journal, April 2016


Dynamics of single mRNP nucleocytoplasmic transport and export through the nuclear pore in living cells
journal, May 2010

  • Mor, Amir; Suliman, Shimrit; Ben-Yishay, Rakefet
  • Nature Cell Biology, Vol. 12, Issue 6
  • DOI: 10.1038/ncb2056

A Saturated FG-Repeat Hydrogel Can Reproduce the Permeability Properties of Nuclear Pore Complexes
journal, August 2007


Promiscuous Binding of Karyopherin β 1 Modulates FG Nucleoporin Barrier Function and Expedites NTF2 Transport Kinetics
journal, February 2015

  • Wagner, Raphael S.; Kapinos, Larisa E.; Marshall, Neil J.
  • Biophysical Journal, Vol. 108, Issue 4
  • DOI: 10.1016/j.bpj.2014.12.041

Considerations and Challenges in Studying Liquid-Liquid Phase Separation and Biomolecular Condensates
journal, January 2019


Surface Properties Determining Passage Rates of Proteins through Nuclear Pores
journal, June 2018


Phase behaviour of disordered proteins underlying low density and high permeability of liquid organelles
journal, June 2017

  • Wei, Ming-Tzo; Elbaum-Garfinkle, Shana; Holehouse, Alex S.
  • Nature Chemistry, Vol. 9, Issue 11
  • DOI: 10.1038/nchem.2803

Nuclear protein import in permeabilized mammalian cells requires soluble cytoplasmic factors
journal, September 1990


Single Molecule Study of the Intrinsically Disordered FG-Repeat Nucleoporin 153
journal, October 2011


Ultrafast microfluidic mixer with three-dimensional flow focusing for studies of biochemical kinetics
journal, January 2010

  • Gambin, Yann; Simonnet, Claire; VanDelinder, Virginia
  • Lab Chip, Vol. 10, Issue 5
  • DOI: 10.1039/B914174J

In vivo imaging of labelled endogenous β-actin mRNA during nucleocytoplasmic transport
journal, September 2010


Plasticity of an Ultrafast Interaction between Nucleoporins and Nuclear Transport Receptors
journal, October 2015


A Programmable DNA Origami Platform for Organizing Intrinsically Disordered Nucleoporins within Nanopore Confinement
journal, January 2018


Brownian Dynamics Simulation of Nucleocytoplasmic Transport: A Coarse-Grained Model for the Functional State of the Nuclear Pore Complex
journal, June 2011


Continuous throughput and long-term observation of single-molecule FRET without immobilization
journal, January 2014

  • Tyagi, Swati; VanDelinder, Virginia; Banterle, Niccolò
  • Nature Methods, Vol. 11, Issue 3
  • DOI: 10.1038/nmeth.2809

A physical model describing the interaction of nuclear transport receptors with FG nucleoporin domain assemblies
journal, April 2016


Cohesiveness tunes assembly and morphology of FG nucleoporin domain meshworks – Implications for nuclear pore permeability
journal, October 2013


Nanomechanical Basis of Selective Gating by the Nuclear Pore Complex
journal, October 2007

  • Lim, Roderick Y. H.; Fahrenkrog, Birthe; Köser, Joachim
  • Science, Vol. 318, Issue 5850
  • DOI: 10.1126/science.1145980

Real-Time Single-Molecule Imaging of the Infection Pathway of an Adeno-Associated Virus
journal, November 2001


Inhibition of amyloid-β plaque formation by α-synuclein
journal, June 2015

  • Bachhuber, Teresa; Katzmarski, Natalie; McCarter, Joanna F.
  • Nature Medicine, Vol. 21, Issue 7
  • DOI: 10.1038/nm.3885

Nuclear transport of baculovirus: Revealing the nuclear pore complex passage
journal, January 2012


A Bimodal Distribution of Two Distinct Categories of Intrinsically Disordered Structures with Separate Functions in FG Nucleoporins
journal, April 2010

  • Yamada, Justin; Phillips, Joshua L.; Patel, Samir
  • Molecular & Cellular Proteomics, Vol. 9, Issue 10
  • DOI: 10.1074/mcp.M000035-MCP201

Microfluidic Device for Single-Molecule Experiments with Enhanced Photostability
journal, September 2009

  • Lemke, Edward A.; Gambin, Yann; Vandelinder, Virginia
  • Journal of the American Chemical Society, Vol. 131, Issue 38
  • DOI: 10.1021/ja9027023

Facilitated aggregation of FG nucleoporins under molecular crowding conditions
journal, December 2012

  • Milles, Sigrid; Huy Bui, Khanh; Koehler, Christine
  • EMBO reports, Vol. 14, Issue 2
  • DOI: 10.1038/embor.2012.204

Amyloid-like interactions within nucleoporin FG hydrogels
journal, March 2010

  • Ader, C.; Frey, S.; Maas, W.
  • Proceedings of the National Academy of Sciences, Vol. 107, Issue 14
  • DOI: 10.1073/pnas.0910163107

Simple rules for passive diffusion through the nuclear pore complex
journal, October 2016

  • Timney, Benjamin L.; Raveh, Barak; Mironska, Roxana
  • The Journal of Cell Biology, Vol. 215, Issue 1
  • DOI: 10.1083/jcb.201601004

The molecular mechanism of nuclear transport revealed by atomic-scale measurements
journal, September 2015


Using Single-Particle Tracking to Study Nuclear Trafficking of Viral Genes
journal, October 2004


Investigating molecular crowding within nuclear pores using polarization-PALM
journal, September 2017


Visualizing a one-way protein encounter complex by ultrafast single-molecule mixing
journal, February 2011

  • Gambin, Yann; VanDelinder, Virginia; Ferreon, Allan Chris M.
  • Nature Methods, Vol. 8, Issue 3
  • DOI: 10.1038/nmeth.1568

FUS pathology in ALS is linked to alterations in multiple ALS-associated proteins and rescued by drugs stimulating autophagy
journal, April 2019


The Permeability of Reconstituted Nuclear Pores Provides Direct Evidence for the Selective Phase Model
journal, August 2012


A short linear motif in scaffold Nup145C connects Y-complex with pre-assembled outer ring Nup82 complex
journal, October 2017


Regulation and aggregation of intrinsically disordered peptides
journal, February 2015

  • Levine, Zachary A.; Larini, Luca; LaPointe, Nichole E.
  • Proceedings of the National Academy of Sciences, Vol. 112, Issue 9
  • DOI: 10.1073/pnas.1418155112

Importin-β modulates the permeability of the nuclear pore complex in a Ran-dependent manner
journal, March 2015


Counteracting chemical chaperone effects on the single-molecule  -synuclein structural landscape
journal, July 2012

  • Ferreon, A. C. M.; Moosa, M. M.; Gambin, Y.
  • Proceedings of the National Academy of Sciences, Vol. 109, Issue 44
  • DOI: 10.1073/pnas.1201802109

Minimal nuclear pore complexes define FG repeat domains essential for transport
journal, February 2004

  • Strawn, Lisa A.; Shen, Tianxiang; Shulga, Nataliya
  • Nature Cell Biology, Vol. 6, Issue 3
  • DOI: 10.1038/ncb1097

Karyopherins regulate nuclear pore complex barrier and transport function
journal, September 2017

  • Kapinos, Larisa E.; Huang, Binlu; Rencurel, Chantal
  • The Journal of Cell Biology, Vol. 216, Issue 11
  • DOI: 10.1083/jcb.201702092

The disordered P granule protein LAF-1 drives phase separation into droplets with tunable viscosity and dynamics
journal, May 2015

  • Elbaum-Garfinkle, Shana; Kim, Younghoon; Szczepaniak, Krzysztof
  • Proceedings of the National Academy of Sciences, Vol. 112, Issue 23
  • DOI: 10.1073/pnas.1504822112

Nuclear Pore Complex Is Able to Transport Macromolecules with Diameters of ∼39 nm
journal, February 2002


Effect of polyethylene glycol on the liquid-liquid phase transition in aqueous protein solutions
journal, October 2002

  • Annunziata, O.; Asherie, N.; Lomakin, A.
  • Proceedings of the National Academy of Sciences, Vol. 99, Issue 22
  • DOI: 10.1073/pnas.212507199

A Liquid-to-Solid Phase Transition of the ALS Protein FUS Accelerated by Disease Mutation
journal, August 2015


Artificial nanopores that mimic the transport selectivity of the nuclear pore complex
journal, December 2008

  • Jovanovic-Talisman, Tijana; Tetenbaum-Novatt, Jaclyn; McKenney, Anna Sophia
  • Nature, Vol. 457, Issue 7232
  • DOI: 10.1038/nature07600