Nup2p dynamically associates with the distal regions of the yeast nuclear pore complex.
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.
Dilworth
D J
DJ
Institute for Systems Biology, Seattle, Washington, 98105, USA.
Suprapto
A
A
Padovan
J C
JC
Chait
B T
BT
Wozniak
R W
RW
Rout
M P
MP
Aitchison
J D
JD
D001665:Binding Sites / Q000502:physiology; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D003847:Deoxyglucose / Q000494:pharmacology; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006160:Guanosine Triphosphate / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics; D022022:Nuclear Pore / Q000378:metabolism*; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D018272:Porins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D021381:Protein Transport / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012440:Saccharomyces; D029701:Saccharomyces cerevisiae Proteins; D019810:Sodium Azide / Q000494:pharmacology; D013211:Staphylococcus aureus / Q000235:genetics; D028961:beta Karyopherins; D020931:ran GTP-Binding Protein / Q000378:metabolism
D005656:Fungal Proteins; D006360:Heat-Shock Proteins; D008164:Luminescent Proteins; C400753:NUP2 protein, S cerevisiae; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; D018272:Porins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; C491115:TIR1 protein, S cerevisiae; D028961:beta Karyopherins; D049452:Green Fluorescent Proteins; D006160:Guanosine Triphosphate; D019810:Sodium Azide; D003847:Deoxyglucose; D020931:ran GTP-Binding Protein