Nuclear protein import is inhibited by an antibody to a lumenal epitope of a nuclear pore complex glycoprotein.
Gp210 is a major transmembrane glycoprotein associated with the nuclear pore complex that is suggested to be important for organizing pore complex architecture and assembly. A mouse monoclonal IgG directed against an epitope in the lumenal domain of rat gp210 was expressed in cultured rat cells by microinjection of mRNA prepared from a hybridoma cell line. The expressed IgG, which becomes assembled into a functional antibody in the lumen of the endoplasmic reticulum, bound to the nuclear envelope in vivo. Expression of anti-gp210 antibody in interphase cells specifically reduced approximately fourfold the mediated nuclear import of a microinjected nuclear protein (nucleoplasmin) coupled to gold particles. The antibody also significantly decreased nuclear influx of a 10-kD dextran by passive diffusion. This transport inhibition did not result from removal of pore complexes from nuclear membranes or from gross alterations in pore complex structure, as shown by EM and immunocytochemistry. A physiological consequence of this transport inhibition was inhibition of cell progression from G2 into M phase. Hence, binding of this antibody to the lumenal side of gp210 must have a transmembrane effect on the structure and functions of the pore complex. These data argue that gp210 is directly or indirectly connected to pore complex constituents involved in mediated import and passive diffusion.
Department of Cell Biology, Scripps Research Institute, La Jolla, California 92037.
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Gerace
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D004259:DNA-Directed DNA Polymerase / Q000032:analysis / Q000378:metabolism; D000939:Epitopes / Q000276:immunology; D016195:G2 Phase; D007074:Immunoglobulin G / Q000235:genetics; D008562:Membrane Glycoproteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008938:Mitosis; D008970:Molecular Weight; D009685:Nuclear Envelope / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007074:Immunoglobulin G; D008562:Membrane Glycoproteins; D009687:Nuclear Proteins; D012333:RNA, Messenger; D004259:DNA-Directed DNA Polymerase