Inhibition of nuclear accumulation of karyophilic proteins in living cells by microinjection of the lectin wheat germ agglutinin.
The lectin wheat germ agglutinin (WGA), which has been reported to inhibit nuclear protein uptake in vitro by isolated nuclei (Finlay et al. (1987) J. Cell Biol. 104, 189), also blocks, on microinjection into living cells, the migration of proteins into the cell nucleus. Radioactively labeled nuclear proteins were injected into the cytoplasm of Xenopus oocytes and their reentry into the nucleus was analyzed in the presence or absence of WGA by two-dimensional gel electrophoresis. In another set of experiments, fluorescently labeled nucleoplasmin was injected, alone or together with WGA, into the cytoplasm of rat hepatoma cells, and its nucleocytoplasmic distribution was studied by quantitative laser fluorescence microscopy. The results indicate that WGA inhibits the uptake of karyophilic proteins in general, independent of their sizes. Since the nucleocytoplasmic flux of a dextran with Mr 10,000 was not affected it can be excluded that WGA acts by a general blockade or constriction of the functional pore channel. At reduced WGA concentrations, the rate but not the final extent of nuclear protein accumulation was decreased. These findings support the concept that the O-glycosidically bound carbohydrates of certain nuclear pore complex proteins are exposed to the pore interior and that these regions are probably involved in nucleocytoplasmic translocation processes.
Dabauvalle
M C
MC
Institute of Zoology I, University of Würzburg, Federal Republic of Germany.
Schulz
B
B
Scheer
U
U
Peters
R
R
D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D008845:Microinjections; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism*; D056489:Nucleoplasmins; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D010750:Phosphoproteins; D014909:Wheat Germ Agglutinins / Q000494:pharmacology*; D014982:Xenopus laevis
D009687:Nuclear Proteins; D056489:Nucleoplasmins; D010750:Phosphoproteins; D014909:Wheat Germ Agglutinins