A quantitative map of nuclear pore assembly reveals two distinct mechanisms
A quantitative map of nuclear pore assembly reveals two distinct mechanisms
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 eukaryotes1,2,3,4. 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.
575–581
Otsuka, Shotaro
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Tempkin, Jeremy O.B.
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Zhang, Wanlu
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Politi, Antonio Z.
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Rybina, Arina
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Hossain, M. Julius
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Kueblbeck, Moritz
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Callegari, Andrea
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Koch, Birgit
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Morero, Natalia Rosalia
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Sali, Andrej
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Ellenberg, Jan
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19 January 2023
Otsuka, Shotaro
f379d77f-b076-43cf-9359-b177f5a2015a
Tempkin, Jeremy O.B.
11e9377a-8b24-41d1-84ed-630cb21bfcd5
Zhang, Wanlu
93ba36d4-5605-4f27-9597-c41e60bd7d0b
Politi, Antonio Z.
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Rybina, Arina
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Hossain, M. Julius
bba1b875-7604-462b-a55b-ba0b54f728e8
Kueblbeck, Moritz
21b89290-812b-44e0-b46f-24e8456b934d
Callegari, Andrea
83bb8749-a25a-482d-9cb0-1f8e128d759c
Koch, Birgit
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Morero, Natalia Rosalia
b0efbc0e-75cc-4874-bdf8-a61c895d032e
Sali, Andrej
204ff034-66ba-4252-be00-4d2184692079
Ellenberg, Jan
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Otsuka, Shotaro, Tempkin, Jeremy O.B., Zhang, Wanlu, Politi, Antonio Z., Rybina, Arina, Hossain, M. Julius, Kueblbeck, Moritz, Callegari, Andrea, Koch, Birgit, Morero, Natalia Rosalia, Sali, Andrej and Ellenberg, Jan
(2023)
A quantitative map of nuclear pore assembly reveals two distinct mechanisms.
Nature, 613 (7944), .
(doi:10.1038/s41586-022-05528-w).
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 eukaryotes1,2,3,4. 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.
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s41586-022-05528-w
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Accepted/In Press date: 4 November 2022
e-pub ahead of print date: 4 January 2023
Published date: 19 January 2023
Additional Information:
Funding Information:
We thank the EMBL Advanced Light Microscopy Facility (ALMF) for their support in STED super-resolution microscopy; S. Schnorrenberg for help with 3D STED; A. Brunner for help with automated live-cell imaging; B. Webb for help with IMP and J. Baker-Lepain for managing the Wynton computer cluster at QBI@UCSF. This work was supported by grants from the Baden Wuerttemberg Foundation (J.E. and A.S.), NIH/NIGMS R01GM083960 (A.S.), NIH/NIGMS P41GM109824 (A.S.), NIH/NIGMS R01GM112108 (A.S.) and the European Molecular Biology Laboratory (EMBL; S.O., A.Z.P., A.R., M.J.H., M.K., A.C., B.K. and J.E.). S.O. and A.R. were further supported by the EMBL Interdisciplinary Postdoc Programme (EIPOD) under Marie Curie Actions COFUND. S.O. was additionally supported by a Japan Society for the Promotion of Science fellowship (postdoctoral fellowship for research abroad).
Publisher Copyright:
© 2023, The Author(s).
Identifiers
Local EPrints ID: 473994
URI: http://eprints.soton.ac.uk/id/eprint/473994
ISSN: 0028-0836
PURE UUID: 92f8be71-6e8e-4eb8-8939-3d024310efac
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Date deposited: 08 Feb 2023 17:32
Last modified: 17 Mar 2024 04:12
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Contributors
Author:
Shotaro Otsuka
Author:
Jeremy O.B. Tempkin
Author:
Wanlu Zhang
Author:
Antonio Z. Politi
Author:
Arina Rybina
Author:
M. Julius Hossain
Author:
Moritz Kueblbeck
Author:
Andrea Callegari
Author:
Birgit Koch
Author:
Natalia Rosalia Morero
Author:
Andrej Sali
Author:
Jan Ellenberg
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