Edgecombe M, Patel R, Whitaker M
Department of Physiology, University College London, UK.
EMBO J. 1991 Dec;10(12):3769-75. doi: 10.1002/j.1460-2075.1991.tb04946.x.
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.
细胞周期调控蛋白p34cdc2的活性在多种细胞类型中受到翻译后调控。利用抗磷酸酪氨酸抗体,我们发现海胆卵受精时,p34cdc2导向的酪氨酸激酶活性增加,导致细胞周期早期p34上磷酸酪氨酸逐渐积累。进入有丝分裂时,p34上的磷酸酪氨酸消失,当胚胎进入下一个细胞周期时,磷酸酪氨酸重新积累。用氯化铵孤雌激活卵时也观察到类似模式。用酪氨酸激酶抑制剂染料木黄酮处理胚胎时,酪氨酸磷酸化和磷酸化/去磷酸化循环受到抑制。另一方面,用放线菌酮抑制蛋白质合成时,该循环持续存在,表明它独立于另一类细胞周期调控蛋白——细胞周期蛋白的合成。用佛波酯(佛波醇肉豆蔻酸酯乙酸酯)进行的额外实验表明,仅激活未受精卵中的蛋白质合成不会刺激p34cdc2酪氨酸激酶活性。我们的结果表明,p34酪氨酸磷酸化循环由受精时的钙瞬变触发。第一个循环独立于受精时的细胞内pH信号,这证实了在海胆胚胎中,该循环与真核细胞分裂周期的一个显著特征——细胞周期蛋白丰度的循环没有紧密耦合。