PT  - JOURNAL ARTICLE
AU  - Shotaro Otsuka
AU  - Jeremy O. B. Tempkin
AU  - Wanlu Zhang
AU  - Antonio Z. Politi
AU  - Arina Rybina
AU  - M. Julius Hossain
AU  - Moritz Kueblbeck
AU  - Andrea Callegari
AU  - Birgit Koch
AU  - Andrej Sali
AU  - Jan Ellenberg
TI  - A quantitative map of nuclear pore assembly reveals two distinct mechanisms
AID  - 10.1101/2021.05.17.444137
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2021.05.17.444137
4099  - http://biorxiv.org/content/early/2022/05/11/2021.05.17.444137.short
4100  - http://biorxiv.org/content/early/2022/05/11/2021.05.17.444137.full
AB  - Understanding how the nuclear pore complex (NPC) assembles is of fundamental importance to grasp the mechanisms behind its essential function and understand its role during evolution of eukaryotes1–4. While we know that at least two NPC assembly pathways exist, one during exit from mitosis and one during nuclear growth in interphase, we currently lack a quantitative map of their molecular events. Here, we use fluorescence correlation spectroscopy (FCS) calibrated live imaging of endogenously fluorescently-tagged nucleoporins to map the changes in 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 employ strikingly different molecular mechanisms, inverting the order of addition of two large structural components, the central ring complex and nuclear filaments. Our dynamic stoichiometry data allows us to perform the first computational simulation that predicts the structure of postmitotic NPC assembly intermediates.Competing Interest StatementThe authors have declared no competing interest.