title: Nuclear pore scaffold structure analyzed by super-resolution microscopy and particle averaging.
abstract: Much of life's essential molecular machinery consists of large protein assemblies that currently pose challenges for structure determination. A prominent example is the nuclear pore complex (NPC), for which the organization of its individual components remains unknown. By combining stochastic super-resolution microscopy, to directly resolve the ringlike structure of the NPC, with single particle averaging, to use information from thousands of pores, we determined the average positions of fluorescent molecular labels in the NPC with a precision well below 1 nanometer. Applying this approach systematically to the largest building block of the NPC, the Nup107-160 subcomplex, we assessed the structure of the NPC scaffold. Thus, light microscopy can be used to study the molecular organization of large protein complexes in situ in whole cells.
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Szymborska
authors.forename: Alex
authors.initials: A
authors.lastname: de Marco
authors.forename: Nathalie
authors.initials: N
authors.lastname: Daigle
authors.forename: Volker C
authors.initials: VC
authors.lastname: Cordes
authors.forename: John A G
authors.initials: JA
authors.lastname: Briggs
authors.forename: Jan
authors.initials: J
authors.lastname: Ellenberg
mesh_terms: D045744:Cell Line, Tumor; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D008853:Microscopy / Q000379:methods*; D018613:Microscopy, Confocal / Q000379:methods; D053758:Nanoparticles / Q000737:chemistry; D015530:Nuclear Matrix / Q000648:ultrastructure*; D022022:Nuclear Pore / Q000648:ultrastructure*; D028861:Nuclear Pore Complex Proteins / Q000737:chemistry* / Q000276:immunology; D010316:Particle Size; D061905:Single-Domain Antibodies / Q000737:chemistry
chemical_list: D005456:Fluorescent Dyes; D028861:Nuclear Pore Complex Proteins; D061905:Single-Domain Antibodies