Nuclear import of glycoconjugates is distinct from the classical NLS pathway.
The nuclear import of many proteins depends on a short peptide sequence called the nuclear localization signal. However, glycosylated proteins, which lack such a nuclear localization signal, upon their injection into the cytosol by electroporation, enter the nucleus in a sugar-dependent manner. This paper brings new insights on the mechanism of this process, based on a study of neoglycoprotein nuclear uptake by digitonin-permeabilized cells. The nuclear import of neoglycoproteins is energy dependent: it does not occur when cells are maintained at 4 degrees C or when cells are ATP-depleted by treatment with apyrase. The nuclear import of neoglycoproteins occurs through the nuclear pore: it is inhibited by preincubation of cells with wheat germ agglutinin, a lectin which binds the nuclear pore glycoproteins and blocks the translocation step of nuclear localization signal bearing proteins through the nuclear pore. Furthermore, the nuclear import of neoglycoproteins does not use the pathway of nuclear localization signal bearing proteins: nuclear import of nuclear localization signal bearing proteins depends on cytosolic factors and is inhibited by treatment of cells with N-ethylmaleimide, while the nuclear import of neoglycoproteins neither requires added cytosolic factors nor is sensitive to alkylation by N-ethylmaleimide. In addition, upon incubation in the presence of a large excess of nuclear localization signal bearing protein, the nuclear import of neoglycoproteins is not inhibited.
Duverger
E
E
Centre de Biophysique Moléculaire, Centre National de la Recherche Scientifique, Orléans, France.
Pellerin-Mendes
C
C
Mayer
R
R
Roche
A C
AC
Monsigny
M
M
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004072:Digitonin; D018274:Electroporation; D005033:Ethylmaleimide / Q000494:pharmacology; D006001:Glycoconjugates / Q000378:metabolism*; D006020:Glycopeptides / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D006367:HeLa Cells; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis; D012261:Ribonucleoproteins / Q000378:metabolism
D006001:Glycoconjugates; D006020:Glycopeptides; D006023:Glycoproteins; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D009842:Oligopeptides; D012261:Ribonucleoproteins; D000255:Adenosine Triphosphate; D004072:Digitonin; D005033:Ethylmaleimide