The yeast nucleoporin Nup53p specifically interacts with Nic96p and is directly involved in nuclear protein import.
The bidirectional nucleocytoplasmic transport of macromolecules is mediated by the nuclear pore complex (NPC) which, in yeast, is composed of approximately 30 different proteins (nucleoporins). Pre-embedding immunogold-electron microscopy revealed that Nic96p, an essential yeast nucleoporin, is located about the cytoplasmic and the nuclear periphery of the central channel, and near or at the distal ring of the yeast NPC. Genetic approaches further implicated Nic96p in nuclear protein import. To more specifically explore the potential role of Nic96p in nuclear protein import, we performed a two-hybrid screen with NIC96 as the bait against a yeast genomic library to identify transport factors and/or nucleoporins involved in nuclear protein import interacting with Nic96p. By doing so, we identified the yeast nucleoporin Nup53p, which also exhibits multiple locations within the yeast NPC and colocalizes with Nic96p in all its locations. Whereas Nup53p is directly involved in NLS-mediated protein import by its interaction with the yeast nuclear import receptor Kap95p, it appears not to participate in NES-dependent nuclear export.
Fahrenkrog
B
B
M. E. Müller Institute for Structural Biology, Biozentrum, University of Basel, CH-4056 Basel, Switzerland. Birthe.Fahrenkrog@embl-heidelberg.de
Hübner
W
W
Mandinova
A
A
Panté
N
N
Keller
W
W
Aebi
U
U
D021581:Active Transport, Cell Nucleus; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D008565:Membrane Proteins; D016253:Microscopy, Immunoelectron; D009154:Mutation; D019913:Nuclear Localization Signals; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism*; D018272:Porins / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics / Q000378:metabolism*; D028961:beta Karyopherins
D005656:Fungal Proteins; D008565:Membrane Proteins; C082533:NIC96 protein, S cerevisiae; C116843:NUP53 protein, S cerevisiae; D019913:Nuclear Localization Signals; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; D018272:Porins; D029701:Saccharomyces cerevisiae Proteins; D028961:beta Karyopherins