title: Domain topology of the p62 complex within the 3-D architecture of the nuclear pore complex.
abstract: The nuclear pore complex (NPC) is the only known gateway for exchange of macromolecules between the cytoplasm and nucleus of eukaryotic cells. One key compound of the NPC is the p62 subcomplex, which consists of the nucleoporins p62, p54, and p58/p45 and is supposed to be involved in nuclear protein import and export. Here we show the localization of distinct domains of the p62 complex by immuno-electron microscopy using isolated nuclei from Xenopus oocytes. To determine the exact position of the p62 complex, we examined the localization of the C and N-terminal domains of p62 by immunogold-labeling using domain-specific antibodies against p62. In addition we expressed epitope-tagged versions of p62, p54, and p58 in Xenopus oocytes and localized the domains with antibodies against the tags. This first systematic analysis of the domain topology of the p62 complex within the NPC revealed that the p62 complex is anchored to the cytoplasmic face of the NPC most likely by the coiled-coil domains of the three nucleoporins. Furthermore, we found the phenylalanine-glycine (FG)-repeat domain of p62, but not of p58 and p54, to be of mobile and flexible nature.
authors.affiliation: M.E. Müller Institute for Structural Biology, Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056 Basel, Switzerland.
authors.forename: Kyrill
authors.initials: K
authors.lastname: Schwarz-Herion
authors.forename: Bohumil
authors.initials: B
authors.lastname: Maco
authors.forename: Ursula
authors.initials: U
authors.lastname: Sauder
authors.forename: Birthe
authors.initials: B
authors.lastname: Fahrenkrog
mesh_terms: D000818:Animals; D000939:Epitopes / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D021621:Imaging, Three-Dimensional; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure*; D046529:Microscopy, Electron, Transmission; D008958:Models, Molecular; D022022:Nuclear Pore / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D028861:Nuclear Pore Complex Proteins; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D014982:Xenopus laevis / Q000378:metabolism
chemical_list: D000939:Epitopes; D008562:Membrane Glycoproteins; D028861:Nuclear Pore Complex Proteins; D011994:Recombinant Proteins; C049312:nuclear pore protein p62