Visualizing the disordered nuclear transport machinery in situ
© 2023. The Author(s)..
The approximately 120 MDa mammalian nuclear pore complex (NPC) acts as a gatekeeper for the transport between the nucleus and cytosol1. The central channel of the NPC is filled with hundreds of intrinsically disordered proteins (IDPs) called FG-nucleoporins (FG-NUPs)2,3. Although the structure of the NPC scaffold has been resolved in remarkable detail, the actual transport machinery built up by FG-NUPs-about 50 MDa-is depicted as an approximately 60-nm hole in even highly resolved tomograms and/or structures computed with artificial intelligence4-11. Here we directly probed conformations of the vital FG-NUP98 inside NPCs in live cells and in permeabilized cells with an intact transport machinery by using a synthetic biology-enabled site-specific small-molecule labelling approach paired with highly time-resolved fluorescence microscopy. Single permeabilized cell measurements of the distance distribution of FG-NUP98 segments combined with coarse-grained molecular simulations of the NPC allowed us to map the uncharted molecular environment inside the nanosized transport channel. We determined that the channel provides-in the terminology of the Flory polymer theory12-a 'good solvent' environment. This enables the FG domain to adopt expanded conformations and thus control transport between the nucleus and cytoplasm. With more than 30% of the proteome being formed from IDPs, our study opens a window into resolving disorder-function relationships of IDPs in situ, which are important in various processes, such as cellular signalling, phase separation, ageing and viral entry.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:617 |
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| Enthalten in: |
Nature - 617(2023), 7959 vom: 26. Mai, Seite 162-169 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Yu, Miao [VerfasserIn] |
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| Links: |
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| Themen: |
Intrinsically Disordered Proteins |
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| Anmerkungen: |
Date Completed 17.05.2023 Date Revised 11.12.2023 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1038/s41586-023-05990-0 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 520 | |a The approximately 120 MDa mammalian nuclear pore complex (NPC) acts as a gatekeeper for the transport between the nucleus and cytosol1. The central channel of the NPC is filled with hundreds of intrinsically disordered proteins (IDPs) called FG-nucleoporins (FG-NUPs)2,3. Although the structure of the NPC scaffold has been resolved in remarkable detail, the actual transport machinery built up by FG-NUPs-about 50 MDa-is depicted as an approximately 60-nm hole in even highly resolved tomograms and/or structures computed with artificial intelligence4-11. Here we directly probed conformations of the vital FG-NUP98 inside NPCs in live cells and in permeabilized cells with an intact transport machinery by using a synthetic biology-enabled site-specific small-molecule labelling approach paired with highly time-resolved fluorescence microscopy. Single permeabilized cell measurements of the distance distribution of FG-NUP98 segments combined with coarse-grained molecular simulations of the NPC allowed us to map the uncharted molecular environment inside the nanosized transport channel. We determined that the channel provides-in the terminology of the Flory polymer theory12-a 'good solvent' environment. This enables the FG domain to adopt expanded conformations and thus control transport between the nucleus and cytoplasm. With more than 30% of the proteome being formed from IDPs, our study opens a window into resolving disorder-function relationships of IDPs in situ, which are important in various processes, such as cellular signalling, phase separation, ageing and viral entry | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
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| 700 | 1 | |a Heidari, Maziar |e verfasserin |4 aut | |
| 700 | 1 | |a Mikhaleva, Sofya |e verfasserin |4 aut | |
| 700 | 1 | |a Tan, Piau Siong |e verfasserin |4 aut | |
| 700 | 1 | |a Mingu, Sara |e verfasserin |4 aut | |
| 700 | 1 | |a Ruan, Hao |e verfasserin |4 aut | |
| 700 | 1 | |a Reinkemeier, Christopher D |e verfasserin |4 aut | |
| 700 | 1 | |a Obarska-Kosinska, Agnieszka |e verfasserin |4 aut | |
| 700 | 1 | |a Siggel, Marc |e verfasserin |4 aut | |
| 700 | 1 | |a Beck, Martin |e verfasserin |4 aut | |
| 700 | 1 | |a Hummer, Gerhard |e verfasserin |4 aut | |
| 700 | 1 | |a Lemke, Edward A |e verfasserin |4 aut | |
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