title: Reconstitution of Nup157 and Nup145N into the Nup84 complex.
abstract: About 30 different nucleoporins (Nups) constitute the nuclear pore complex. We have affinity-purified 28 of these nuclear pore proteins and identified new nucleoporin interactions by this analysis. We found that Nup157 and Nup170, two members of the large structural Nups, and the Gly-Leu-Phe-Gly nucleoporin Nup145N specifically co-purified with members of the Nup84 complex. In addition, Nup145N co-enriched during Nup157 purification. By in vitro reconstitution, we demonstrate that Nup157 and Nup145N form a nucleoporin subcomplex. Moreover, we show that Nup157 and Nup145N bind to the heptameric Nup84 complex. This assembly thus represents approximately one-third of all nucleoporins. To characterize Nup157 structurally, we purified and analyzed it by electron microscopy. Nup157 is a hollow sphere that resembles a clamp or a gripping hand. Thus, we could reconstitute an interaction between a large structural Nup, an FG repeat Nup, and a major structural module of the nuclear pore complex.
authors.affiliation: Biochemie-Zentrum der Universität Heidelberg, Im Neuenheimer Feld 328, 69120 Heidelberg, Germany.
authors.forename: Malik
authors.initials: M
authors.lastname: Lutzmann
authors.forename: Ruth
authors.initials: R
authors.lastname: Kunze
authors.forename: Karin
authors.initials: K
authors.lastname: Stangl
authors.forename: Philipp
authors.initials: P
authors.lastname: Stelter
authors.forename: Katalin Fejes
authors.initials: KF
authors.lastname: Tóth
authors.forename: Bettina
authors.initials: B
authors.lastname: Böttcher
authors.forename: Ed
authors.initials: E
authors.lastname: Hurt
mesh_terms: D019281:Dimerization; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D025941:Protein Interaction Mapping; D040901:Proteomics / Q000379:methods; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: C088627:NUP145 protein, S cerevisiae; C097179:NUP157 protein, S cerevisiae; C097894:NUP84 protein, S cerevisiae; D028861:Nuclear Pore Complex Proteins; D029701:Saccharomyces cerevisiae Proteins