Nup180, a novel nuclear pore complex protein localizing to the cytoplasmic ring and associated fibrils.
Using an autoimmune serum from a patient with overlap connective tissue disease we have identified by biochemical and immunocytochemical approaches an evolutionarily conserved nuclear pore complex (NPC) protein with an estimated molecular mass of 180 kD and an isoelectric point of approximately 6.2 which we have designated as nup180. Extraction of isolated nuclear envelopes with 2 M urea and chromatography of the solubilized proteins on WGA-Sepharose demonstrated that nup180 is a peripheral membrane protein and does not react with WGA. Affinity-purified antibodies yielded a punctate immunofluorescent pattern of the nuclear surface of mammalian cells and stained brightly the nuclear envelope of cryosectioned Xenopus oocytes. Nuclei reconstituted in vitro in Xenopus egg extract were also stained in the characteristic punctate fashion. Immunogold EM localized nup180 exclusively to the cytoplasmic ring of NPCs and short fibers emanating therefrom into the cytoplasm. Antibodies to nup180 did not inhibit nuclear protein transport in vivo nor in vitro. Despite the apparent lack of involvement in NPC assembly or nucleocytoplasmic transport processes, the conservation of nup180 across species and its exclusive association with the NPC cytoplasmic ring suggests an important, though currently undefined function for this novel NPC protein.
Wilken
N
N
Department of Cell and Developmental Biology, Theodor-Boveri-Institute, University of Würzburg, Germany.
Kossner
U
U
Senécal
J L
JL
Scheer
U
U
Dabauvalle
M C
MC
D000818:Animals; D000918:Antibody Specificity; D001323:Autoantibodies / Q000276:immunology; D001324:Autoantigens / Q000737:chemistry / Q000378:metabolism; D001693:Biological Transport, Active; D003240:Connective Tissue Diseases / Q000276:immunology; D003593:Cytoplasm / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D007526:Isoelectric Point; D008970:Molecular Weight; D009685:Nuclear Envelope / Q000648:ultrastructure*; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D009865:Oocytes / Q000648:ultrastructure; D014982:Xenopus laevis
D001323:Autoantibodies; D001324:Autoantigens; C079377:NUP153 protein, human; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins