PT  - JOURNAL ARTICLE
AU  - Vilma Jimenez Sabinina
AU  - M. Julius Hossain
AU  - Jean-Karim Hériché
AU  - Philipp Hoess
AU  - Bianca Nijmeijer
AU  - Shyamal Mosalaganti
AU  - Moritz Kueblbeck
AU  - Andrea Callegari
AU  - Anna Szymborska
AU  - Martin Beck
AU  - Jonas Ries
AU  - Jan Ellenberg
TI  - 3D super-resolution fluorescence microscopy maps the variable molecular architecture of the Nuclear Pore Complex
AID  - 10.1101/2020.11.27.386599
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2020.11.27.386599
4099  - http://biorxiv.org/content/early/2021/04/16/2020.11.27.386599.short
4100  - http://biorxiv.org/content/early/2021/04/16/2020.11.27.386599.full
AB  - Nuclear pore complexes (NPCs) are large macromolecular machines that mediate the traffic between the nucleus and the cytoplasm. In vertebrates, each NPC consists of ~1000 proteins, termed nucleoporins, and has a mass of over 100 MDa. While a pseudo-atomic static model of the central scaffold of the NPC has recently been assembled by integrating data from isolated proteins and complexes, many structural components still remain elusive due to the enormous size and flexibility of the NPC. Here, we explored the power of 3D super-resolution microscopy combined with computational classification and averaging to explore the 3D structure of the NPC in single human cells. We show that this approach can build the first integrated 3D structural map containing both central as well as peripheral NPC subunits with molecular specificity and nanoscale resolution. Our unbiased classification of over ten thousand individual NPCs indicates that the nuclear ring and the nuclear basket can adopt different conformations. Our approach opens up the exciting possibility to relate different structural states of the NPC to function in situ.Competing Interest StatementThe authors have declared no competing interest.