Characterization of a major nucleoplasmin-like germinal vesicle protein which is rapidly phosphorylated before germinal vesicle breakdown in Spisula solidissima.

Oocytes of the surf clam, Spisula solidissima, are arrested at the G2/M boundary of meiotic prophase I. At this stage, they possess a prominent germinal vesicle (GV), comprising about 25% of the total oocyte volume, in which pools of mRNAs and proteins that facilitate the rapid rounds of cell division which occur early in development are stored. We have isolated and characterized an abundant 49-kDa phosphoprotein, localized exclusively to the GV, which shares properties with nucleoplasmin. Like nucleoplasmin, this 49-kDa protein is a heat-stable, highly acidic phosphoprotein [containing over 32% (Glx + Asx) with an isoelectric point of about 4.0] and is soluble in 80% ammonium sulfate. In contrast to the pentameric nucleoplasmin, much of this protein is isolated as a disulfide-linked multimer which migrates at 120 kDa. In addition, a fraction of the 49-kDa protein is associated via disulfide bonds to a doublet of 42-kDa proteins. In vivo, this 49-kDa protein is phosphorylated prior to germinal vesicle breakdown (GVBD) and at 5 min after oocyte activation rapidly incorporates 32P to 60% of maximal level, suggesting that this protein plays a major role in the cascade of events which lead to oocyte activation and GVBD.