Authors:	Yu, Miao Heidari, Maziar Mikhaleva, Sofya Tan, Piau Siong Mingu, Sara Ruan, Hao Reinkemeier, Christopher D. Obarska-Kosinska, Agnieszka Siggel, Marc Beck, Martin Hummer, Gerhard Lemke, Edward A.
Title:	Visualizing the disordered nuclear transport machinery in situ
Online publication date:	21-Nov-2023
Year of first publication:	2023
Language:	english
Abstract:	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 intelligence4,5,6,7,8,9,10,11. 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.
DDC:	570 Biowissenschaften 570 Life sciences
Institution:	Johannes Gutenberg-Universität Mainz
Department:	FB 10 Biologie
Place:	Mainz
ROR:	https://ror.org/023b0x485
DOI:	http://doi.org/10.25358/openscience-9682
Version:	Published version
Publication type:	Zeitschriftenaufsatz
Document type specification:	Scientific article
License:	CC BY
Information on rights of use:	https://creativecommons.org/licenses/by/4.0/
Journal:	Nature 617
Pages or article number:	162 169
Publisher:	Nature Publ. Group
Publisher place:	London u.a.
Issue date:	2023
ISSN:	1476-4687 0028-0836
Publisher DOI:	10.1038/s41586-023-05990-0
Appears in collections:	JGU-Publikationen