A quantitative map of nuclear pore assembly reveals two distinct mechanisms

Shotaro Otsuka (), Jeremy O. B. Tempkin, Wanlu Zhang, Antonio Z. Politi, Arina Rybina, M. Julius Hossain, Moritz Kueblbeck, Andrea Callegari, Birgit Koch, Natalia Rosalia Morero, Andrej Sali and Jan Ellenberg ()
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Shotaro Otsuka: European Molecular Biology Laboratory
Jeremy O. B. Tempkin: University of California, San Francisco
Wanlu Zhang: European Molecular Biology Laboratory
Antonio Z. Politi: European Molecular Biology Laboratory
Arina Rybina: European Molecular Biology Laboratory
M. Julius Hossain: European Molecular Biology Laboratory
Moritz Kueblbeck: European Molecular Biology Laboratory
Andrea Callegari: European Molecular Biology Laboratory
Birgit Koch: European Molecular Biology Laboratory
Natalia Rosalia Morero: European Molecular Biology Laboratory
Andrej Sali: University of California, San Francisco
Jan Ellenberg: European Molecular Biology Laboratory

Nature, 2023, vol. 613, issue 7944, 575-581

Abstract: Abstract Understanding how the nuclear pore complex (NPC) is assembled is of fundamental importance to grasp the mechanisms behind its essential function and understand its role during the evolution of eukaryotes1–4. There are at least two NPC assembly pathways—one during the exit from mitosis and one during nuclear growth in interphase—but we currently lack a quantitative map of these events. Here we use fluorescence correlation spectroscopy calibrated live imaging of endogenously fluorescently tagged nucleoporins to map the changes in the composition and stoichiometry of seven major modules of the human NPC during its assembly in single dividing cells. This systematic quantitative map reveals that the two assembly pathways have distinct molecular mechanisms, in which the order of addition of two large structural components, the central ring complex and nuclear filaments are inverted. The dynamic stoichiometry data was integrated to create a spatiotemporal model of the NPC assembly pathway and predict the structures of postmitotic NPC assembly intermediates.

Date: 2023
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DOI: 10.1038/s41586-022-05528-w

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