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タイトル | A checkpoint function for Nup98 in nuclear pore formation suggested by novel inhibitory nanobodies. |
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ジャーナル・号・ページ | EMBO J, Vol. 43, Issue 11, Page 2198-2232, Year 2024 |
掲載日 | 2024年4月22日 |
![]() | Mireia Solà Colom / Zhenglin Fu / Philip Gunkel / Thomas Güttler / Sergei Trakhanov / Vasundara Srinivasan / Kathrin Gregor / Tino Pleiner / Dirk Görlich / ![]() ![]() |
PubMed 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 ...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. |
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手法 | EM (単粒子) / X線回折 |
解像度 | 1.41 - 4.4 Å |
構造データ | EMDB-14849: Nup93 in complex with 5-Nup93 inhibitory NB and 15-Nup93 tracking NB ![]() PDB-7now: ![]() PDB-7nqa: ![]() PDB-8cds: ![]() PDB-8cdt: ![]() PDB-8ozb: |
化合物 | ![]() ChemComp-NA: ![]() ChemComp-SO4: ![]() ChemComp-HOH: ![]() ChemComp-TRS: ![]() ChemComp-EDO: ![]() ChemComp-PEG: ![]() ChemComp-GOL: |
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