Xenopus nucleoplasmin: egg vs. oocyte.
Nucleoplasmin has been purified from either oocytes or unfertilized eggs of the frog, Xenopus laevis. We find that the pentameric form of egg nucleoplasmin exhibits an apparent molecular mass approximately 15 000 daltons larger than its oocyte counterpart upon sodium dodecyl sulfate (SDS)-acrylamide gel electrophoresis. Egg nucleoplasmin monomers are more heterogeneous, substantially more acidic, and overall larger in apparent molecular weight than oocyte nucleoplasmin monomers when analyzed by isoelectric focusing or SDS gel electrophoresis. Protease digestions indicate that the structural differences between egg and oocyte nucleoplasmin are primarily confined to the N-terminal halves of the proteins. The structural diversity observed is accompanied by a difference in the ability of nucleoplasmin from the two sources to act as a nucleosome assembly agent in vitro. Egg nucleoplasmin efficiently promotes the formation of nucleosomes onto circular pBR322 DNA in vitro at physiological ionic strength and at physiological histone:DNA ratios, while oocyte nucleoplasmin is markedly deficient in serving as an in vitro chromatin assembly agent under all conditions which we have tested. Treatment of egg nucleoplasmin in vitro with alkaline phosphatase demonstrates that the structural diversity between egg and oocyte nucleoplasmin results primarily from extensive additional phosphorylation of the egg protein. The relevance of nucleoplasmin phosphorylation in leading to differences in the chromatin assembly activity of this protein both in vitro and in vivo is considered.
D000818:Animals; D000911:Antibodies, Monoclonal; D002843:Chromatin / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D046911:Macromolecular Substances; D008970:Molecular Weight; D009687:Nuclear Proteins; D056489:Nucleoplasmins; D009698:Nucleoproteins / Q000302:isolation & purification*; D009865:Oocytes / Q000032:analysis*; D010063:Ovum / Q000032:analysis*; D010446:Peptide Fragments / Q000032:analysis; D010750:Phosphoproteins; D010957:Plasmids; D014357:Trypsin; D014981:Xenopus
D000911:Antibodies, Monoclonal; D002843:Chromatin; D046911:Macromolecular Substances; D009687:Nuclear Proteins; D056489:Nucleoplasmins; D009698:Nucleoproteins; D010446:Peptide Fragments; D010750:Phosphoproteins; D014357:Trypsin