supplement- The Mobile FG Nucleoporin Nup98 Is a Cofactor for Crm1-dependent Protein Export

  • Oka M
  • Asally M
  • Yasuda Y
 et al. 
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Abstract

Nup98 is a mobile nucleoporin that forms distinct dots in the nucleus, and, although a role for Nup98 in nuclear transport has been suggested, its precise function remains unclear. Here, we show that Nup98 plays an important role in Crm1-mediated nuclear protein export. Nuclear, but not cytoplasmic, dots of EGFP-tagged Nup98 disappeared rapidly after cell treatment with leptomycin B, a specific inhibitor of the nuclear export receptor, Crm1. Mutational analysis demonstrated that Nup98 physically and functionally interacts with Crm1 in a RanGTP-dependent manner through its N-terminal phenylalanine-glycine (FG) repeat region. Moreover, the activity of the Nup98-Crm1 complex was modulated by RanBP3, a known cofactor for Crm1-mediated nuclear export. Finally, cytoplasmic microinjection of anti-Nup98 inhibited the Crm1-dependent nuclear export of proteins, concomitant with the accumulation of anti-Nup98 in the nucleus. These results clearly demonstrate that Nup98 functions as a novel shuttling cofactor for Crm1-mediated nuclear export in conjunction with RanBP3.

Author-supplied keywords

  • Active Transport
  • Animals
  • Cell Nucleolus
  • Cell Nucleolus: metabolism
  • Cell Nucleus
  • Cell Nucleus: physiology
  • Cytoplasmic and Nuclear
  • Cytoplasmic and Nuclear: genetics
  • Cytoplasmic and Nuclear: metabolism
  • DNA Mutational Analysis
  • Dactinomycin
  • Dactinomycin: metabolism
  • HeLa Cells
  • Humans
  • Karyopherins
  • Karyopherins: genetics
  • Karyopherins: metabolism
  • Mice
  • NIH 3T3 Cells
  • Nuclear Pore Complex Proteins
  • Nuclear Pore Complex Proteins: genetics
  • Nuclear Pore Complex Proteins: metabolism
  • Nuclear Proteins
  • Nuclear Proteins: genetics
  • Nuclear Proteins: metabolism
  • Nucleocytoplasmic Transport Proteins
  • Nucleocytoplasmic Transport Proteins: genetics
  • Nucleocytoplasmic Transport Proteins: metabolism
  • Protein Synthesis Inhibitors
  • Protein Synthesis Inhibitors: metabolism
  • Receptors
  • Recombinant Fusion Proteins
  • Recombinant Fusion Proteins: genetics
  • Recombinant Fusion Proteins: metabolism
  • ran GTP-Binding Protein
  • ran GTP-Binding Protein: genetics
  • ran GTP-Binding Protein: metabolism

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Authors

  • Masahiro Oka

  • Munehiro Asally

  • Yoshinari Yasuda

  • Yutaka Ogawa

  • Taro Tachibana

  • Yoshihiro Yoneda

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