title: Snapshots of nuclear pore complexes in action captured by cryo-electron tomography.
abstract: Nuclear pore complexes reside in the nuclear envelope of eukaryotic cells and mediate the nucleocytoplasmic exchange of macromolecules. Traffic is regulated by mobile transport receptors that target their cargo to the central translocation channel, where phenylalanine-glycine-rich repeats serve as binding sites. The structural analysis of the nuclear pore is a formidable challenge given its size, its location in a membranous environment and its dynamic nature. Here we have used cryo-electron tomography to study the structure of nuclear pore complexes in their functional environment, that is, in intact nuclei of Dictyostelium discoideum. A new image-processing strategy compensating for deviations of the asymmetric units (protomers) from a perfect eight-fold symmetry enabled us to refine the structure and to identify new features. Furthermore, the superposition of a large number of tomograms taken in the presence of cargo, which was rendered visible by gold nanoparticles, has yielded a map outlining the trajectories of import cargo. Finally, we have performed single-molecule Monte Carlo simulations of nuclear import to interpret the experimentally observed cargo distribution in the light of existing models for nuclear import.
authors.affiliation: Max Planck Institute of Biochemistry, D-82152 Martinsried, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Beck
authors.forename: Vladan
authors.initials: V
authors.lastname: Lucić
authors.forename: Friedrich
authors.initials: F
authors.lastname: Förster
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Baumeister
authors.forename: Ohad
authors.initials: O
authors.lastname: Medalia
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D020285:Cryoelectron Microscopy; D004023:Dictyostelium / Q000166:cytology* / Q000648:ultrastructure*; D008958:Models, Molecular; D008968:Molecular Conformation; D022022:Nuclear Pore / Q000378:metabolism* / Q000648:ultrastructure*; D014054:Tomography