Journal Article

Architecture of the symmetric core of the nuclear pore

Lin, Daniel H., Fan, Yanbin, Schilbach, Sandra, Perriches, Thibaud, Davenport, Andrew M., Collins, Leslie N., Thierbach, Karsten, Hoelz, André, Stuwe, Tobias, Jeon, Young E., Mobbs, George, Rundlet, Emily J., Huber, Ferdinand M.

2016

Overview

Nuclear pore complexes (NPCs) consist of around 1000 protein subunits, are embedded in the membrane that surrounds the nucleus, and regulate transport between the nucleus and the cytoplasm. Although the overall shape of NPCs is known, the details of this macromolecular complex have been obscure. Now, Lin et al. have reconstituted the pore components, determined the interactions between them, and fitted them into a tomographic reconstruction. Kosinski et al. have provided an architectural map of the inner ring of the pore. Science , this issue pp. 10.1126/science.aaf1015 and 363 Reconstitution, spectroscopy, and crystallography allow the construction of a model of the human nuclear pore. The nuclear pore complex (NPC) controls the transport of macromolecules between the nucleus and cytoplasm, but its molecular architecture has thus far remained poorly defined. We biochemically reconstituted NPC core protomers and elucidated the underlying protein-protein interaction network. Flexible linker sequences, rather than interactions between the structured core scaffold nucleoporins, mediate the assembly of the inner ring complex and its attachment to the NPC coat. X-ray crystallographic analysis of these scaffold nucleoporins revealed the molecular details of their interactions with the flexible linker sequences and enabled construction of full-length atomic structures. By docking these structures into the cryoelectron tomographic reconstruction of the intact human NPC and validating their placement with our nucleoporin interactome, we built a composite structure of the NPC symmetric core that contains ~320,000 residues and accounts for ~56 megadaltons of the NPC’s structured mass. Our approach provides a paradigm for the structure determination of similarly complex macromolecular assemblies.

• Publisher: American Association for the Advancement of Science,The American Association for the Advancement of Science,AAAS
• Subject: Active Transport, Cell Nucleus / Amino Acid Sequence / Architecture / Biochemistry / Cryoelectron Microscopy / Crystal structure / Crystallography, X-Ray / Cytology / Cytoplasm / Cytoplasm - metabolism / Electron Microscope Tomography / Fungal Proteins - chemistry / Fungal Proteins - genetics / Fungal Proteins - metabolism / Gene expression / Humans / Molecular Sequence Data / Nuclear Pore - chemistry / Nuclear Pore - metabolism / Nuclear Pore - ultrastructure / Nuclear Pore Complex Proteins - chemistry / Nuclear Pore Complex Proteins - genetics / Nuclear Pore Complex Proteins - metabolism / Protein Interaction Maps / Protein Structure, Tertiary / Protein Subunits - chemistry / Protein Subunits - genetics / Protein Subunits - metabolism / RESEARCH ARTICLE SUMMARY / Scientific Concepts / Symmetry