Removal of a single pore subcomplex results in vertebrate nuclei devoid of nuclear pores

Mol Cell. 2003 Apr;11(4):853-64. doi: 10.1016/s1097-2765(03)00116-3.


The vertebrate nuclear pore complex, 30 times the size of a ribosome, assembles from a library of soluble subunits and two membrane proteins. Using immunodepletion of Xenopus nuclear reconstitution extracts, it has previously been possible to assemble nuclei lacking pore subunits tied to protein import, export, or mRNA export. However, these altered pores all still possessed the bulk of pore structure. Here, we immunodeplete a single subunit, the Nup107-160 complex, using antibodies to Nup85 and Nup133, two of its components. The resulting reconstituted nuclei are severely defective for NLS import and DNA replication. Strikingly, they show a profound defect for every tested nucleoporin. Even the integral membrane proteins POM121 and gp210 are absent or unorganized. Scanning electron microscopy reveals pore-free nuclei, while addback of the Nup107-160 complex restores functional pores. We conclude that the Nup107-160 complex is a pivotal determinant for vertebrate nuclear pore complex assembly.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Active Transport, Cell Nucleus / genetics*
  • Animals
  • DNA Replication / genetics*
  • Female
  • HeLa Cells
  • Humans
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Microscopy, Electron, Scanning
  • Minor Histocompatibility Antigens
  • Nuclear Pore / metabolism*
  • Nuclear Pore / ultrastructure
  • Nuclear Pore Complex Proteins / deficiency*
  • Nuclear Pore Complex Proteins / genetics
  • Nuclear Pore Complex Proteins / metabolism
  • Nuclear Proteins*
  • Oocytes / metabolism
  • Platelet Membrane Glycoproteins / genetics
  • Platelet Membrane Glycoproteins / metabolism
  • Protein Subunits / deficiency*
  • Protein Subunits / genetics
  • Saccharomyces cerevisiae Proteins / genetics
  • Vertebrates / metabolism*
  • Xenopus Proteins
  • Xenopus laevis


  • GP 210 protein, human
  • Membrane Proteins
  • Minor Histocompatibility Antigens
  • NUP107 protein, human
  • NUP133 protein, human
  • NUP160 protein, human
  • NUP85 protein, S cerevisiae
  • Nuclear Pore Complex Proteins
  • Nuclear Proteins
  • Nup160 protein, Xenopus
  • Platelet Membrane Glycoproteins
  • Pom121 protein, rat
  • Protein Subunits
  • Saccharomyces cerevisiae Proteins
  • Xenopus Proteins