Reversible inhibition of protein import into the nucleus by wheat germ agglutinin injected into cultured cells.
The importance of glycoproteins located in the nuclear envelope in nuclear transport was tested by microinjection of karyophilic proteins into the cytoplasm of cultured human cells together with various lectins. Wheat germ agglutinin (WGA) blocked the nuclear transport of nucleoplasmin, a nuclear protein of Xenopus laevis oocytes, and of nonnuclear proteins conjugated with a synthetic peptide containing the nuclear localization signal sequence for simian virus 40 (SV40) large T antigen. Its inhibitory activity persisted for about 1 h after its injection into the cells and then gradually decreased. Export of at least some kinds of RNA from the nucleus seemed not to be affected by WGA even when import of the proteins into the nucleus was completely blocked (within 1 h after WGA injection). Moreover, WGA did not inhibit the passive diffusion of fluorescein isothiocyanate (FITC)-dextran (average Mr 17,900) into the nucleus. Wistaria floribunda agglutinin (WFA), concanavalin A (Con A), and lentil lectin did not block nuclear transport. These results indicate that WGA specifically blocks active protein import, but not passive diffusion of materials into the nucleus.
D000952:Antigens, Polyomavirus Transforming / Q000378:metabolism*; D001667:Binding, Competitive / Q000187:drug effects*; D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D003593:Cytoplasm / Q000378:metabolism; D004058:Diffusion; D006801:Humans; D008845:Microinjections; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000378:metabolism*; D056489:Nucleoplasmins; D010750:Phosphoproteins; D021382:Protein Sorting Signals / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis; D014909:Wheat Germ Agglutinins / Q000378:metabolism / Q000494:pharmacology*
D000952:Antigens, Polyomavirus Transforming; D009687:Nuclear Proteins; D056489:Nucleoplasmins; D010750:Phosphoproteins; D021382:Protein Sorting Signals; D014909:Wheat Germ Agglutinins; D012313:RNA