title: Distinct association of the nuclear pore protein Nup153 with A- and B-type lamins.
abstract: The nuclear envelope (NE) is a double membrane physical barrier, which separates the nucleus from the cytoplasm. Underlying the NE are the nuclear lamins, which in combination with inner nuclear membrane proteins form the lamina. The lamina is crucial for maintaining the structural integrity of the nucleus and for positioning of nuclear pore complexes (NPCs) within the NE. The nucleoporin Nup153 has previously been reported to bind to B-type lamins. However, the specificity of this interaction is not well established. Here we show that Nup153 exhibits multiple binding sites for A- and B-type lamins. Using GST-pull down assays, we found that both the N-terminal domain of Nup153 and its C terminus associate with the Ig-fold domain of A- and B-type lamins. By employing purified Nup153 and lamin proteins in blot overlay assays we revealed that both the N-terminal and the C-terminal domain of Nup153 are directly interacting with the lamins. Moreover, we provide evidence that mutations in the lamin A Ig-fold domain selectively affect Nup153-binding, suggesting that Nup153 may play a role in lamin-associated diseases, known as laminopathies. Together our results indicate a far more intricate interplay between Nup153 and nuclear lamins than previously accepted.
authors: M.E. Mueller Institute for Structural Biology, Biozentrum, University of Basel, Switzerland.
authors: Teiba
authors: T
authors: Al-Haboubi
authors: Dale K
authors: DK
authors: Shumaker
authors: Joachim
authors: J
authors: Köser
authors: Manfred
authors: M
authors: Wehnert
authors: Birthe
authors: B
authors: Fahrenkrog
mesh_terms: D001665:Binding Sites; D002478:Cells, Cultured; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D034904:Lamin Type A / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D034921:Lamin Type B / Q000737:chemistry / Q000378:metabolism*; D028861:Nuclear Pore Complex Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D034904:Lamin Type A; D034921:Lamin Type B; C079377:NUP153 protein, human; D028861:Nuclear Pore Complex Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase