Molecular Cell, Год журнала: 2023, Номер 83(18), С. 3232 - 3233
Опубликована: Сен. 1, 2023
Язык: Английский
Cell, Год журнала: 2024, Номер 187(19), С. 5267 - 5281.e13
Опубликована: Авг. 9, 2024
The nuclear pore complex (NPC) is the sole mediator of nucleocytoplasmic transport. Despite great advances in understanding its conserved core architecture, peripheral regions can exhibit considerable variation within and between species. One such structure cage-like basket. crucial roles mRNA surveillance chromatin organization, an architectural has remained elusive. Using in-cell cryo-electron tomography subtomogram analysis, we explored NPC's structural variations basket across fungi (yeast; S. cerevisiae), mammals (mouse; M. musculus), protozoa (T. gondii). integrative modeling, computed a model yeast that revealed how hub nucleoporins (Nups) ring binds to basket-forming Mlp/Tpr proteins: coiled-coil domains form struts basket, while their unstructured termini constitute distal densities, which potentially serve as docking site for preprocessing before
Язык: Английский
Процитировано
27
Nature, Год журнала: 2025, Номер unknown
Опубликована: Март 19, 2025
The nuclear pore complex (NPC) mediates nucleocytoplasmic exchange, catalysing a massive flux of protein and nucleic acid material in both directions1. Distinct trafficking pathways for import export would be an elegant solution to avoid unproductive collisions opposing movements. However, the three-dimensional (3D) nanoscale spatiotemporal dynamics macromolecules traversing NPC remains challenging visualize on timescale millisecond-scale transport events. Here we used 3D MINFLUX2 identify scaffold then simultaneously monitor export, thereby establishing that processes occur overlapping regions central pore. Whereas translocation-arrested complexes bound at periphery, tracks translocating within region revealed preference approximately 40- 50-nm diameter annulus with minimal circumferential movement, indicating activity-dependent confinement permeability barrier. Movement was 1,000-fold slower than interspersed pauses, highly restricted environment structural constraints and/or transient binding events during transport. These results demonstrate high precision reduced photobleaching is major advantage MINFLUX tracking, barrier divided into annular rings distinct functional properties.
Язык: Английский
Процитировано
1
Analytical Chemistry, Год журнала: 2024, Номер 96(26), С. 10765 - 10771
Опубликована: Июнь 21, 2024
The nuclear pore complex (NPC) is a proteinaceous nanopore that solely and selectively regulates the molecular transport between cytoplasm nucleus of eukaryotic cell. ∼50 nm-diameter NPC perforates double-membrane envelope to mediate both passive facilitated transport, thereby playing paramount biological biomedical roles. Herein, we visualize single NPCs by scanning electrochemical microscopy (SECM). high spatial resolution accomplished employing ∼25 ion-selective nanopipets monitor tetrabutylammonium at individual NPCs. SECM images are quantitatively analyzed finite element method confirm this work represents highest-resolution nanoscale imaging samples. Significantly, apply powerful technique address long-debated origin central plug NPC. Nanoscale demonstrates unplugged more permeable small probe ion than plugged This result supports hypothesis not an intrinsic transporter, but impermeable macromolecule, e.g., ribonucleoprotein, trapped in nanopore. Moreover, also mechanism where divided into pathway for RNA export peripheral protein import efficiently bidirectional traffic.
Язык: Английский
Процитировано
6
Nature Cell Biology, Год журнала: 2024, Номер 26(9), С. 1504 - 1519
Опубликована: Авг. 13, 2024
The nuclear basket attaches to the nucleoplasmic side of pore complex (NPC), coupling transcription mRNA quality control and export. expands functional repertoire a subset NPCs in Saccharomyces cerevisiae by drawing unique RNA/protein interactome. Yet, how docks onto NPC core remains unknown. By integrating AlphaFold-based interaction screens, electron microscopy membrane-templated reconstitution, we uncovered membrane-anchored tripartite junction between core. subunit Nup60 harbours three adjacent short linear motifs, which connect Mlp1, parallel homodimer consisting coiled-coil segments interrupted flexible hinges, Nup85 Y-complex. We reconstituted Y-complex•Nup60•Mlp1 assembly on synthetic membrane validated protein interfaces vivo. Here explain motif-based can substantially reshape structure function, advancing our understanding compositional conformational heterogeneity.
Язык: Английский
Процитировано
6
bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown
Опубликована: Янв. 2, 2024
The Nuclear Pore Complex (NPC) facilitates rapid and selective nucleocytoplasmic transport of molecules as large ribosomal subunits viral capsids. It is not clear how key emergent properties this arise from the system components their interactions. To address question, we constructed an integrative coarse-grained Brownian dynamics model through a single NPC, followed by coupling it with kinetic Ran-dependent in entire cell. microscopic parameters were fitted to reflect experimental data theoretical information regarding transport, without making any assumptions about its properties. resulting reductionist validated reproducing several features used for construction, such morphology central transporter, rates passive facilitated diffusion function size valency, situ radial distributions pre-ribosomal subunits, active suggests that NPC functions essentially virtual gate whose flexible phenylalanine-glycine (FG) repeat proteins raise entropy barrier pore. Importantly, core functionality greatly enhanced design features, including 'fuzzy' transient interactions, multivalency, redundancy copy number FG nucleoporins, exponential kinetics thermodynamics accordance transition state theory, energy-reliant RanGTP concentration gradient. These result robust resilient rate selectivity wide array cargo ranging few kilodaltons megadaltons size. By dissecting these our provides quantitative starting point rationally modulating artificial mimics.
Язык: Английский
Процитировано
5
bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown
Опубликована: Март 28, 2024
The nuclear pore complex (NPC) is the sole mediator of nucleocytoplasmic transport. Despite great advances in understanding its conserved core architecture, peripheral regions can exhibit considerable variation within and between species. One such structure cage-like basket. crucial roles mRNA surveillance chromatin organization, an architectural has remained elusive. Using in-cell cryo-electron tomography subtomogram analysis, we explored NPC's structural variations basket across fungi (yeast;
Язык: Английский
Процитировано
3
QRB Discovery, Год журнала: 2025, Номер 6
Опубликована: Янв. 1, 2025
Abstract Integrative modeling enables structure determination for large macromolecular assemblies by combining data from multiple experiments with theoretical and computational predictions. Recent advancements in AI-based prediction cryo electron-microscopy have sparked renewed enthusiasm integrative modeling; structures methods can be integrated situ maps to characterize assemblies. This approach previously allowed us others determine the architectures of diverse assemblies, such as nuclear pore complexes, chromatin remodelers, cell–cell junctions. Experimental spanning several scales was used these studies, ranging high-resolution data, X-ray crystallography AlphaFold structure, low-resolution cryo-electron tomography co-immunoprecipitation experiments. Two recurrent challenges emerged across a range studies. First, contained significant fractions disordered regions, necessitating development new regions context ordered regions. Second, needed developed utilize information tomography, timely challenge structural biology is increasingly moving towards characterization. Here, we recapitulate recent developments proteins analysis highlight other opportunities method modeling.
Язык: Английский
Процитировано
0
Nucleus, Год журнала: 2025, Номер 16(1)
Опубликована: Июнь 4, 2025
Due to its pivotal role as a regulator of nucleocytoplasmic transport, the structure and dynamic gating mechanism nuclear pore complex (NPC) is subject immense interest. Here, we report key recent advancements discussed at Selective Transport Control in Biological Biomimetic Nanopores meeting (Monte Verità, Switzerland, 2024) that gathered NPC experts from range disciplines. Novel insights were reported cutting-edge super-resolution techniques enable direct interrogation NPC's central transporter; computational models unravel mechanisms selective barrier; synthetic mimics valuable vitro for delineating permeability transport dynamics. Altogether, three major highlighted: (i) presence dynamically organised pathways within NPC, (ii) receptors enrich reinforce barrier, (iii) ability DNA origami nanostructures mimic aspects with unprecedented precision. Overall, marked convergence our understanding function by unraveling nanoscale.
Язык: Английский
Процитировано
0
Current Opinion in Structural Biology, Год журнала: 2024, Номер 87, С. 102841 - 102841
Опубликована: Май 24, 2024
Язык: Английский
Процитировано
2
Molecular & Cellular Proteomics, Год журнала: 2024, Номер 23(5), С. 100758 - 100758
Опубликована: Апрель 2, 2024
The acceleration of advances in proteomics has enabled integration with imaging at the EM and LM levels, cryo-electron microscopy protein structures, artificial intelligence (AI) proteins comprehensively accurately resolved for cell structures nm to sub-nm resolution. Proteomics continues outpace experimentally based structural imaging, but their ultimate is a path towards goal compendium all understand mechanistically structure function.
Язык: Английский
Процитировано
1