Localization of the Ran-GTP binding protein RanBP2 at the cytoplasmic side of the nuclear pore complex.
A partial cDNA clone coding for the mouse homologue of the human Ran-GTP binding protein, RanBP2, has been isolated by screening of a murine expression library with antibodies to nup180, a previously identified nuclear pore complex protein (nucleoporin). Whether the antibodies cross-reacted with the polypeptide encoded by the cDNA clone or, alternatively, nup180 is proteolytically related to RanBP2, has not been determined. The 3795-bp open reading frame of the cDNA encodes a polypeptide consisting of 1265 amino acids with three Ran-GTP binding domains (RanBD) that are almost identical with published partial amino acid sequences of human RanBP2 as deduced from several partial cDNA clones of other authors. Sequence analysis further revealed that murine RanBP2 contains tandemly repeated zinc fingers of Cys2-Cys2 type and multiple copies of the FXFG nucleoporin "signature" motif clustered in regions preceding the RanBDs. Antibodies raised against a synthetic peptide of the derived amino acid sequence decorated the cytoplasmic rings of nuclear pore complexes (NPCs) as shown by immunogold electron microscopy. We suggest that the cytoplasmically disposed nucleoporin RanBP2 provides docking sites for import substrate-receptor complexes and, further, that the affinity of these sites to the transport substrate is modulated in a Ran-dependent fashion.
Wilken
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N
Department of Cell and Developmental Biology, Theodor-Boveri-Institute, University of Würzburg, Germany.
Senécal
J L
JL
Scheer
U
U
Dabauvalle
M C
MC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000737:chemistry*; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000235:genetics; D006801:Humans; D051379:Mice; D018832:Molecular Chaperones; D008969:Molecular Sequence Data; D009685:Nuclear Envelope / Q000737:chemistry*; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D016335:Zinc Fingers; D020931:ran GTP-Binding Protein
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D018832:Molecular Chaperones; C079377:NUP153 protein, human; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; C513084:Nup153 protein, mouse; C513083:Nup153 protein, rat; C094493:ran-binding protein 2; D020931:ran GTP-Binding Protein