A cyclin-abundance cycle-independent p34cdc2 tyrosine phosphorylation cycle in early sea urchin embryos.
The activity of the cell cycle control protein p34cdc2 is post-translationally regulated in a variety of cell types. Using anti-phosphotyrosine antibodies, we find that p34cdc2-directed tyrosine kinase activity increases at fertilization in sea urchin eggs, leading to a gradual accumulation of phosphotyrosine on p34 during the early part of the cell cycle. Loss of phosphotyrosine from p34 accompanies entry into mitosis and phosphotyrosine reaccumulates as the embryo enters the next cell cycle. A similar pattern is seen when eggs are parthenogenetically activated with ammonium chloride. Tyrosine phosphorylation and phosphorylation/dephosphorylation cycles are suppressed when embryos are treated with the tyrosine kinase inhibitor genistein. On the other hand, a cycle persists when protein synthesis is inhibited with emetine, indicating that it is independent of the synthesis of another class of cell cycle control proteins, the cyclins. Additional experiments with the phorbol ester, phorbol myristate acetate, demonstrate that activating protein synthesis alone in unfertilized eggs does not result in stimulation of p34cdc2 tyrosine kinase activity. Our results indicate that p34 tyrosine phosphorylation cycles are triggered by the fertilization Cai transient. The first cycle is independent of the fertilization pHi signal, confirming that, in sea urchin embryos, the cycle is not tightly coupled to the cycle of cyclin abundance that is a prominent feature of the eukaryotic cell division cycle.
Edgecombe
M
M
Department of Physiology, University College London, UK.
Patel
R
R
Whitaker
M
M
D000818:Animals; D015153:Blotting, Western; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D002453:Cell Cycle; D003429:Cross Reactions; D016213:Cyclins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004640:Emetine / Q000494:pharmacology; D019833:Genistein; D007529:Isoflavones / Q000494:pharmacology; D008938:Mitosis; D010063:Ovum / Q000166:cytology / Q000201:enzymology; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012617:Sea Urchins / Q000196:embryology; D014443:Tyrosine / Q000378:metabolism
D016213:Cyclins; D007529:Isoflavones; D014443:Tyrosine; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D016203:CDC2 Protein Kinase; D004640:Emetine