Structural analysis of a metazoan nuclear pore complex reveals a fused concentric ring architecture.
The sole gateway for molecular exchange between the cytoplasm and the nucleus is the nuclear pore complex (NPC). This large supramolecular assembly mediates transport of cargo into and out of the nucleus and fuse the inner and outer nuclear membranes to form an aqueous translocation channel. The NPC is composed of eight proteinaceous asymmetric units forming a pseudo-8-fold symmetric passage. Due to its shear size, complexity, and plastic nature, dissecting the high-resolution three-dimensional structure of the NPC in its hydrated state is a formidable challenge. Toward this goal, we applied cryo-electron tomography to spread nuclear envelopes from Xenopus oocytes. To compensate for perturbations of the 8-fold symmetry of individual NPCs, we performed symmetry-independent asymmetric unit averaging of three-dimensional tomographic NPC volumes to eventually yield a refined model at 6.4 nm resolution. This approach revealed novel structural features, particularly in the spoke-ring complex and luminal domains. Fused concentric ring architecture of the spoke-ring complex was found along the translocation channel. Additionally, a comparison of the refined Xenopus model to that of its Dictyostelium homologue yielded similar pore diameters at the level of the three canonical rings, although the Xenopus NPC was found to be 30% taller than the Dictyostelium pore. This discrepancy is attributed primarily to the relatively low homology and different organization of some nucleoporins in the Dictyostelium genome as compared to that of vertebrates. Nevertheless, the experimental conditions impose a preferred axial orientation of the NPCs within spread Xenopus oocyte nuclear envelopes. This may at least in part explain the increased height of the reconstructed vertebrate NPCs compared to those obtained from tomographic reconstruction of intact Dictyostelium nuclei.
Frenkiel-Krispin
Daphna
D
Max-Planck-Institute for Biochemistry, D-82152 Martinsried, Germany.
Maco
Bohumil
B
Aebi
Ueli
U
Medalia
Ohad
O
D000818:Animals; D020285:Cryoelectron Microscopy; D004023:Dictyostelium; D005260:Female; D021621:Imaging, Three-Dimensional; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000737:chemistry / Q000648:ultrastructure; D022022:Nuclear Pore / Q000737:chemistry* / Q000648:ultrastructure*; D009865:Oocytes / Q000737:chemistry / Q000648:ultrastructure; D020836:Protein Structure, Quaternary; D021122:Protein Subunits; D013045:Species Specificity; D014054:Tomography; D014982:Xenopus laevis
D046912:Multiprotein Complexes; D021122:Protein Subunits