Yeast nucleoporins involved in passive nuclear envelope permeability

Nataliya Shulga, Nima Mosammaparast, Richard Wozniak, David S. Goldfarb

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84 Scopus citations

Abstract

The vertebrate nuclear pore complex (NPC) harbors an ~ 10-nm diameter diffusion channel that is large enough to admit 50-kD polypeptides. We have analyzed the permeability properties of the Saccharomyces cerevisiae nuclear envelope (NE) using import (NLS) and export (NES) signal-containing green fluorescent protein (GFP) reporters. Compared with wild-type, passive export rates of a classical karyopherin/importin (Kap) Kap60p/Kap95p-targeted NLS- GFP reporter (cNLS-GFP) were significantly faster in nup188-Δ and nup170-Δ cells. Similar results were obtained using two other NLS-GFP reporters, containing either the Kap104p-targeted Nab2p NLS (rgNLS) or the Kap121p- targeted Pho4p NLS (pNLS). Elevated levels of Hsp70 stimulated cNLS-GFP import, but had no effect on the import of rgNLS-GFP. Thus, the role of Hsp70 in NLS-directed import may be NLS- or targeting pathway-specific. Equilibrium sieving limits for the diffusion channel were assessed in vivo using NES-GFP reporters of 36-126 kD and were found to be greater than wild-type in nup188- Δ and nup170-Δ cells. We propose that Nup170p and Nup188p are involved in establishing the functional resting diameter of the NPC's central transport channel.

Original languageEnglish
Pages (from-to)1027-1038
Number of pages12
JournalJournal of Cell Biology
Volume149
Issue number5
DOIs
StatePublished - May 29 2000

Keywords

  • Diffusion channel
  • Green fluorescent protein
  • Import/export signals
  • Nuclear pore complex
  • Saccharomyces cerevisiae

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