A checkpoint function for Nup98 in nuclear pore formation suggested by novel 
inhibitory nanobodies

Solà Colom, Mireia; Fu, Zhenglin; Gunkel, Philip; Güttler, Thomas; Trakhanov, Sergei; Srinivasan, Vasundara; Gregor, Kathrin; Pleiner, Tino; Görlich, Dirk

doi:10.1038/s44318-024-00081-w

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A checkpoint function for Nup98 in nuclear pore formation suggested by novel inhibitory nanobodies

MPS-Authors

/persons/resource/persons297996

Solà Colom, Mireia
Department of Cellular Logistics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

/persons/resource/persons297998

Fu, Zhenglin
Department of Cellular Logistics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

/persons/resource/persons267154

Gunkel, Philip
Department of Cellular Logistics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

/persons/resource/persons15158

Güttler, Thomas
Department of Cellular Logistics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

/persons/resource/persons181080

Trakhanov, Sergei
Department of Cellular Logistics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

/persons/resource/persons298000

Srinivasan, Vasundara
Department of Cellular Logistics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

/persons/resource/persons267148

Gregor, Kathrin
Department of Cellular Logistics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

/persons/resource/persons135005

Pleiner, Tino
Department of Cellular Logistics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

/persons/resource/persons15132

Görlich, Dirk
Department of Cellular Logistics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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solà-colom-et-al-2024
(出版社版), 15MB

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引用

Solà Colom, M., Fu, Z., Gunkel, P., Güttler, T., Trakhanov, S., Srinivasan, V., Gregor, K., Pleiner, T., & Görlich, D. (2024). A checkpoint function for Nup98 in nuclear pore formation suggested by novel inhibitory nanobodies. The EMBO Journal. doi:10.1038/s44318-024-00081-w.

引用: https://hdl.handle.net/21.11116/0000-000F-39DE-8

要旨

Nuclear pore complex (NPC) biogenesis is a still enigmatic example of protein self-assembly. We now introduce several cross-reacting anti-Nup nanobodies for imaging intact nuclear pore complexes from frog to human. We also report a simplified assay that directly tracks postmitotic NPC assembly with added fluorophore-labeled anti-Nup nanobodies. During interphase, NPCs are inserted into a pre-existing nuclear envelope. Monitoring this process is challenging because newly assembled NPCs are indistinguishable from pre-existing ones. We overcame this problem by inserting Xenopus-derived NPCs into human nuclear envelopes and using frog-specific anti-Nup nanobodies for detection. We further asked whether anti-Nup nanobodies could serve as NPC assembly inhibitors. Using a selection strategy against conserved epitopes, we obtained anti-Nup93, Nup98, and Nup155 nanobodies that block Nup–Nup interfaces and arrest NPC assembly. We solved structures of nanobody-target complexes and identified roles for the Nup93 α-solenoid domain in recruiting Nup358 and the Nup214·88·62 complex, as well as for Nup155 and the Nup98 autoproteolytic domain in NPC scaffold assembly. The latter suggests a checkpoint linking pore formation to the assembly of the Nup98-dominated permeability barrier.