In situ structural analysis of the human nuclear pore complex
Alexander von Appen,Jan Kosinski,Lenore Sparks,Alessandro Ori,Amanda L. DiGuilio,Benjamin Vollmer,Marie-Therese Mackmull,Niccolò Banterle,Luca Parca,Panagiotis L. Kastritis,Katarzyna Buczak,Shyamal Mosalaganti,Wim J. H. Hagen,Amparo Andres-Pons,Edward A. Lemke,Peer Bork,Wolfram Antonin,Joseph S. Glavy,Khanh Huy Bui,Martin Beck +19 more
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TLDR
This work combines cryo-electron tomography with mass spectrometry, biochemical analysis, perturbation experiments and structural modelling to generate the most comprehensive architectural model of the human nuclear pore complex to date.Abstract:
Nuclear pore complexes are fundamental components of all eukaryotic cells that mediate nucleocytoplasmic exchange. Determining their 110-megadalton structure imposes a formidable challenge and requires in situ structural biology approaches. Of approximately 30 nucleoporins (Nups), 15 are structured and form the Y and inner-ring complexes. These two major scaffolding modules assemble in multiple copies into an eight-fold rotationally symmetric structure that fuses the inner and outer nuclear membranes to form a central channel of ~60 nm in diameter. The scaffold is decorated with transport-channel Nups that often contain phenylalanine-repeat sequences and mediate the interaction with cargo complexes. Although the architectural arrangement of parts of the Y complex has been elucidated, it is unclear how exactly it oligomerizes in situ. Here we combine cryo-electron tomography with mass spectrometry, biochemical analysis, perturbation experiments and structural modelling to generate, to our knowledge, the most comprehensive architectural model of the human nuclear pore complex to date. Our data suggest previously unknown protein interfaces across Y complexes and to inner-ring complex members. We show that the transport-channel Nup358 (also known as Ranbp2) has a previously unanticipated role in Y-complex oligomerization. Our findings blur the established boundaries between scaffold and transport-channel Nups. We conclude that, similar to coated vesicles, several copies of the same structural building block--although compositionally identical--engage in different local sets of interactions and conformations.read more
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The nuclear pore complex: understanding its function through structural insight.
Martin Beck,Ed Hurt +1 more
TL;DR: The recent convergence of crystallographic and biochemical in vitro analysis of nucleoporins, the components of the NPC, with cryo-electron microscopic imaging of the entire NPC in situ has provided first pseudo-atomic view of its central core and revealed that an unexpected network of short linear motifs is an important spatial organization principle.
Journal ArticleDOI
Visualizing the molecular sociology at the HeLa cell nuclear periphery
Julia Mahamid,Stefan Pfeffer,Miroslava Schaffer,Elizabeth Villa,Radostin Danev,Luis Kuhn Cuellar,Friedrich Förster,Anthony A. Hyman,Jürgen M. Plitzko,Wolfgang Baumeister +9 more
TL;DR: Cryo-ET was used to visualize previously elusive structures, such as nucleosome chains and the filaments of the nuclear lamina, in situ, which revealed the native structure and organization of the cytoplasmic translation machinery.
Journal ArticleDOI
Integrative structure and functional anatomy of a nuclear pore complex
Seung Joong Kim,Javier Fernandez-Martinez,Ilona Nudelman,Yi Shi,Wenzhu Zhang,Barak Raveh,Thurston Herricks,Brian D. Slaughter,Joanna A. Hogan,Paula Upla,Ilan E. Chemmama,Riccardo Pellarin,Ignacia Echeverria,Manjunatha Shivaraju,Azraa S. Chaudhury,Junjie Wang,Rosemary Williams,Jay R. Unruh,Charles H. Greenberg,Erica Y. Jacobs,Zhiheng Yu,M. Jason de la Cruz,Roxana Mironska,David L. Stokes,John D. Aitchison,John D. Aitchison,Martin F. Jarrold,Jennifer L. Gerton,Steven J. Ludtke,Christopher W. Akey,Brian T. Chait,Andrej Sali,Michael P. Rout +32 more
TL;DR: The structure of the entire 552-protein nuclear pore complex of the yeast Saccharomyces cerevisiae is determined at sub-nanometre precision by satisfying a wide range of data relating to the molecular arrangement of its constituents.
Journal ArticleDOI
The Nuclear Pore Complex as a Flexible and Dynamic Gate.
TL;DR: The current status of NPC research is reviewed with a focus on the functional implications of its structural and compositional heterogeneity.
Journal ArticleDOI
MINFLUX nanoscopy delivers 3D multicolor nanometer resolution in cells.
Klaus C. Gwosch,Jasmin K. Pape,Francisco Balzarotti,Philipp Hoess,Jan Ellenberg,Jonas Ries,Stefan W. Hell +6 more
TL;DR: It is shown that by localizing individual switchable fluorophores with a probing donut-shaped excitation beam, MINFLUX nanoscopy can provide resolutions in the range of 1 to 3 nm for structures in fixed and living cells.
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