Reconstitution of Nup157 and Nup145N into the Nup84 complex.
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.
Lutzmann
Malik
M
Biochemie-Zentrum der Universität Heidelberg, Im Neuenheimer Feld 328, 69120 Heidelberg, Germany.
Kunze
Ruth
R
Stangl
Karin
K
Stelter
Philipp
P
Tóth
Katalin Fejes
KF
Böttcher
Bettina
B
Hurt
Ed
E
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
C088627:NUP145 protein, S cerevisiae; C097179:NUP157 protein, S cerevisiae; C097894:NUP84 protein, S cerevisiae; D028861:Nuclear Pore Complex Proteins; D029701:Saccharomyces cerevisiae Proteins