Nuclear pore assembly proceeds by an inside-out extrusion of the nuclear envelope

  1. Shotaro Otsuka
  2. Khanh Huy Bui
  3. Martin Schorb
  4. M Julius Hossain
  5. Antonio Z Politi
  6. Birgit Koch
  7. Mikhail Eltsov
  8. Martin Beck
  9. Jan Ellenberg  Is a corresponding author
  1. European Molecular Biology Laboratory, Germany
  2. McGill University, Canada

Abstract

The nuclear pore complex (NPC) mediates nucleocytoplasmic transport through the nuclear envelope. How the NPC assembles into this double membrane boundary has remained enigmatic. Here, we captured temporally staged assembly intermediates by correlating live cell imaging with high-resolution electron tomography and super-resolution microscopy. Intermediates were dome-shaped evaginations of the inner nuclear membrane (INM), that grew in diameter and depth until they fused with the flat outer nuclear membrane. Live and super-resolved fluorescence microscopy revealed the molecular maturation of the intermediates, which initially contained the nuclear and cytoplasmic ring component Nup107, and only later the cytoplasmic filament component Nup358. EM particle averaging showed that the evagination base was surrounded by an 8-fold rotationally symmetric ring structure from the beginning and that a growing mushroom-shaped density was continuously associated with the deforming membrane. Quantitative structural analysis revealed that interphase NPC assembly proceeds by an asymmetric inside-out extrusion of the INM.

Article and author information

Author details

  1. Shotaro Otsuka

    Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3976-0843
  2. Khanh Huy Bui

    Department of Anatomy and Cell Biology, McGill University, Montreal, Canada
    Competing interests
    The authors declare that no competing interests exist.
  3. Martin Schorb

    Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. M Julius Hossain

    Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3303-5755
  5. Antonio Z Politi

    Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  6. Birgit Koch

    Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  7. Mikhail Eltsov

    Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Martin Beck

    Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  9. Jan Ellenberg

    Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    For correspondence
    jan.ellenberg@embl.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5909-701X

Funding

Deutsche Forschungsgemeinschaft (EL 246/3-2)

  • Jan Ellenberg

European Research Council (309271-NPCAtlas)

  • Martin Beck

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Karsten Weis, ETH Zurich, Switzerland

Publication history

  1. Received: June 23, 2016
  2. Accepted: September 13, 2016
  3. Accepted Manuscript published: September 15, 2016 (version 1)
  4. Version of Record published: October 14, 2016 (version 2)

Copyright

© 2016, Otsuka et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 6,684
    Page views
  • 1,614
    Downloads
  • 88
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, Scopus, PubMed Central.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Shotaro Otsuka
  2. Khanh Huy Bui
  3. Martin Schorb
  4. M Julius Hossain
  5. Antonio Z Politi
  6. Birgit Koch
  7. Mikhail Eltsov
  8. Martin Beck
  9. Jan Ellenberg
(2016)
Nuclear pore assembly proceeds by an inside-out extrusion of the nuclear envelope
eLife 5:e19071.
https://doi.org/10.7554/eLife.19071