The nucleoporin Nup153 is required for nuclear pore basket formation, nuclear pore complex anchoring and import of a subset of nuclear proteins.
The nuclear pore complex (NPC) is a large proteinaceous structure through which bidirectional transport of macromolecules across the nuclear envelope (NE) takes place. Nup153 is a peripheral NPC component that has been implicated in protein and RNP transport and in the interaction of NPCs with the nuclear lamina. Here, Nup153 is localized by immunogold electron microscopy to a position on the nuclear ring of the NPC. Nuclear reconstitution is used to investigate the role of Nup153 in nucleo- cytoplasmic transport and NPC architecture. NPCs assembled in the absence of Nup153 lacked several nuclear basket components, were unevenly distributed in the NE and, unlike wild-type NPCs, were mobile within the NE. Importin alpha/beta-mediated protein import into the nucleus was strongly reduced in the absence of Nup153, while transportin-mediated import was unaffected. This was due to a reduction in import complex translocation rather than to defective receptor recycling. Our results therefore reveal functions for Nup153 in NPC assembly, in anchoring NPCs within the NE and in mediating specific nuclear import events.
Walther
T C
TC
European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.
Fornerod
M
M
Pickersgill
H
H
Goldberg
M
M
Allen
T D
TD
Mattaj
I W
IW
D000818:Animals; D002417:Cattle; D005260:Female; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D007150:Immunohistochemistry; D028884:Karyopherins / Q000378:metabolism; D046911:Macromolecular Substances; D008297:Male; D016253:Microscopy, Immunoelectron; D022022:Nuclear Pore / Q000502:physiology* / Q000648:ultrastructure; D028861:Nuclear Pore Complex Proteins / Q000502:physiology*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D056489:Nucleoplasmins; D009865:Oocytes; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012261:Ribonucleoproteins / Q000235:genetics / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000235:genetics / Q000378:metabolism; D014982:Xenopus laevis
D034441:Heterogeneous-Nuclear Ribonucleoproteins; D028884:Karyopherins; D046911:Macromolecular Substances; C079377:NUP153 protein, human; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; D056489:Nucleoplasmins; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; D012261:Ribonucleoproteins; D012710:Serum Albumin, Bovine