Nuclear pore assembly proceeds by an inside-out extrusion of the nuclear envelope
- European Molecular Biology Laboratory, Germany
- 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
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
- Karsten Weis, ETH Zurich, Switzerland
Version history
- Received: June 23, 2016
- Accepted: September 13, 2016
- Accepted Manuscript published: September 15, 2016 (version 1)
- 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.
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Nuclear pore assembly proceeds by an inside-out extrusion of the nuclear envelope
eLife 5:e19071.
https://doi.org/10.7554/eLife.19071
