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Keywords:

  • calmodulin;
  • CaM kinase;
  • fertilization;
  • kinase;
  • p34cdc2;
  • sea urchin

Upon fertilization, the sea urchin egg synthesizes proteins which impart a Ca2+ dependence to M-phase onset. A potential target of this Ca2+ dependence may be CaM kinase-II (the multifunctional [type II] Ca2+/calmodulin [CaM]-dependent protein kinase) which is necessary for nuclear envelope breakdown in fertilized sea urchin eggs. This study was intended to determine whether sea urchin CaMK-II is activated after fertilization and whether it interacts with other known M-phase regulators, such as p34cdc2. We report that total CaMK-II activity, measured by solution assays, increases after fertilization, peaking just prior to cleavage. Interestingly, total CaMK-II activity continues to fluctuate, peaking again prior to second and third cleavage. Gel assays also reveal enhanced levels of the 56 and 62 kDa potential CaMK-II phosphoproteins after fertilization. Finally, CaMK-II activity and only the 62 kDa phosphoprotein physically associate with p34cdc2, but again only after fertilization. These changes in CaMK-II activity and p34cdc2-association after fertilization may ensure that Ca2+ signals are targeted to the M-phase machinery at the appropriate developmental times.