title: Nup214 is required for CRM1-dependent nuclear protein export in vivo.
abstract: Nucleoporins mediate transport of macromolecules across the nuclear pore complex, yet the function of many individual nucleoporins is largely unresolved. To address this question, we depleted cells of the cytoplasmic nucleoporins Nup214/CAN and Nup358/RanBP2 by RNA interference. Depletion of Nup214 resulted in codepletion of its binding partner, Nup88. Nuclear pore complexes assembled in the absence of Nup214/Nup88 or Nup358 were fully functional in nuclear protein import, whereas nuclear mRNA export was slightly impaired. Depletion of Nup358 had only a minor effect on nuclear protein export. In contrast, depletion of Nup214/Nup88 led to strongly reduced CRM1-mediated export of the shuttling transcription factor NFAT as well as a human immunodeficiency virus-Rev derivative. A specific role of Nup214 in protein export is furthered by the biochemical properties of a high-affinity complex containing Nup214, CRM1, RanGTP, and an export cargo. Our results show that the Nup214/Nup88 complex is required for efficient CRM1-mediated transport, supporting a model involving a high-affinity binding site for CRM1 at Nup214 in the terminal steps of export.
authors.affiliation: Universität Göttingen, Zentrum für Biochemie und Molekulare Zellbiologie, Humboldtallee 23, 37073 Göttingen, Germany.
authors.forename: Saskia
authors.initials: S
authors.lastname: Hutten
authors.forename: Ralph H
authors.initials: RH
authors.lastname: Kehlenbach
mesh_terms: D021581:Active Transport, Cell Nucleus; D048708:Cell Growth Processes; D003593:Cytoplasm / Q000378:metabolism; D020690:GTPase-Activating Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D028884:Karyopherins / Q000378:metabolism*; D018832:Molecular Chaperones / Q000378:metabolism; D050778:NFATC Transcription Factors / Q000378:metabolism; D022022:Nuclear Pore / Q000378:metabolism*; D028861:Nuclear Pore Complex Proteins / Q000172:deficiency / Q000378:metabolism*; D011061:Poly A / Q000378:metabolism; D011485:Protein Binding; D034622:RNA Interference; D034443:RNA Transport; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D000097606:Exportin 1 Protein
chemical_list: D020690:GTPase-Activating Proteins; D028884:Karyopherins; D018832:Molecular Chaperones; D050778:NFATC Transcription Factors; D028861:Nuclear Pore Complex Proteins; D011061:Poly A; D018160:Receptors, Cytoplasmic and Nuclear; D000097606:Exportin 1 Protein; C085914:NUP214 protein, human; C104835:NUP88 protein, human; C092317:RANGAP1 protein, human; C094493:ran-binding protein 2