{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Sola Colom M"],"funding":["European Research Council","Boehringer Ingelheim Fonds"],"pagination":["2198-2232"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC11148069"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["43(11)"],"pubmed_abstract":["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."],"journal":["The EMBO journal"],"pubmed_title":["A checkpoint function for Nup98 in nuclear pore formation suggested by novel inhibitory nanobodies."],"pmcid":["PMC11148069"],"funding_grant_id":["StuDySARCOMERE"],"pubmed_authors":["Guttler T","Gorlich D","Gunkel P","Fu Z","Gregor K","Trakhanov S","Srinivasan V","Sola Colom M","Pleiner T"],"additional_accession":[]},"is_claimable":false,"name":"A checkpoint function for Nup98 in nuclear pore formation suggested by novel inhibitory nanobodies.","description":"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.","dates":{"release":"2024-01-01T00:00:00Z","publication":"2024 Jun","modification":"2026-06-02T08:28:49.09Z","creation":"2026-04-16T03:11:36.859Z"},"accession":"S-EPMC11148069","cross_references":{"pubmed":["38649536"],"doi":["10.1038/s44318-024-00081-w"]}}