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Published online 25 June 2001. doi:10.1083/jcb.153.7.1465
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© The Rockefeller University Press, 0021-9525/2001/6/1465/ $5.00
The Journal of Cell Biology, Volume 153, Number 7, June 25, 2001 1465-1478

Original Article

Nup2p Dynamically Associates with the Distal Regions of the Yeast Nuclear Pore Complex

David J. Dilwortha,b, Adisetyantari Supraptoc, Julio C. Padovanc, Brian T. Chaitc, Richard W. Wozniakb, Michael P. Routb, and John D. Aitchisona,b
a Institute for Systems Biology, Seattle, Washington, 98105
b Department of Cell Biology, University of Alberta, Edmonton, Canada, T6G 2H7
c The Rockefeller University, New York, New York, 10021

Correspondence to: John D. Aitchison, Institute for Systems Biology, 4225 Roosevelt Way, Suite 200, Seattle, WA 98105-6099. Tel:(206) 732-1344 Fax:(206) 732-1299 E-mail:jaitchison{at}systemsbiology.org.

Nucleocytoplasmic transport is mediated by the interplay between soluble transport factors and nucleoporins resident within the nuclear pore complex (NPC). Understanding this process demands knowledge of components of both the soluble and stationary phases and the interface between them. Here, we provide evidence that Nup2p, previously considered to be a typical yeast nucleoporin that binds import- and export-bound karyopherins, dynamically associates with the NPC in a Ran-facilitated manner. When bound to the NPC, Nup2p associates with regions corresponding to the nuclear basket and cytoplasmic fibrils. On the nucleoplasmic face, where the Ran–GTP levels are predicted to be high, Nup2p binds to Nup60p. Deletion of NUP60 renders Nup2p nucleoplasmic and compromises Nup2p-mediated recycling of Kap60p/Srp1p. Depletion of Ran–GTP by metabolic poisoning, disruption of the Ran cycle, or in vitro by cell lysis, results in a shift of Nup2p from the nucleoplasm to the cytoplasmic face of the NPC. This mobility of Nup2p was also detected using heterokaryons where, unlike nucleoporins, Nup2p was observed to move from one nucleus to the other. Together, our data support a model in which Nup2p movement facilitates the transition between the import and export phases of nucleocytoplasmic transport.

Key Words: nucleoporin, nuclear transport, karyopherin, importin, exportin


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