Ferrell J E, Wu M, Gerhart J C, Martin G S
Department of Zoology, University of Wisconsin-Madison 53706.
Mol Cell Biol. 1991 Apr;11(4):1965-71. doi: 10.1128/mcb.11.4.1965-1971.1991.
We have examined the time course of protein tyrosine phosphorylation in the meiotic cell cycles of Xenopus laevis oocytes and the mitotic cell cycles of Xenopus eggs. We have identified two proteins that undergo marked changes in tyrosine phosphorylation during these processes: a 42-kDa protein related to mitogen-activated protein kinase or microtubule-associated protein-2 kinase (MAP kinase) and a 34-kDa protein identical or related to p34cdc2. p42 undergoes an abrupt increase in its tyrosine phosphorylation at the onset of meiosis 1 and remains tyrosine phosphorylated until 30 min after fertilization, at which point it is dephosphorylated. p42 also becomes tyrosine phosphorylated after microinjection of oocytes with partially purified M-phase-promoting factor, even in the presence of cycloheximide. These findings suggest that MAP kinase, previously implicated in the early responses of somatic cells to mitogens, is also activated at the onset of meiotic M phase and that MAP kinase can become tyrosine phosphorylated downstream from M-phase-promoting factor activation. We have also found that p34 goes through a cycle of tyrosine phosphorylation and dephosphorylation prior to meiosis 1 and mitosis 1 but is not detectable as a phosphotyrosyl protein during the 2nd through 12th mitotic cell cycles. It may be that the delay between assembly and activation of the cyclin-p34cdc2 complex that p34cdc2 tyrosine phosphorylation provides is not needed in cell cycles that lack G2 phases. Finally, an unidentified protein or group of proteins migrating at 100 to 116 kDa increase in tyrosine phosphorylation throughout maturation, are dephosphorylated or degraded within 10 min of fertilization, and appear to cycle between low-molecular-weight forms and high-molecular-weight forms during early embryogenesis.
我们研究了非洲爪蟾卵母细胞减数分裂细胞周期以及非洲爪蟾卵有丝分裂细胞周期中蛋白质酪氨酸磷酸化的时间进程。我们鉴定出两种在这些过程中酪氨酸磷酸化发生显著变化的蛋白质:一种42 kDa的蛋白质,与丝裂原活化蛋白激酶或微管相关蛋白2激酶(MAP激酶)相关;另一种34 kDa的蛋白质,与p34cdc2相同或相关。p42在减数分裂Ⅰ开始时酪氨酸磷酸化突然增加,并一直保持酪氨酸磷酸化状态,直到受精后30分钟,此时它被去磷酸化。即使在存在环己酰亚胺的情况下,用部分纯化的M期促进因子显微注射卵母细胞后,p42也会发生酪氨酸磷酸化。这些发现表明,之前认为与体细胞对丝裂原的早期反应有关的MAP激酶,在减数分裂M期开始时也被激活,并且MAP激酶可以在M期促进因子激活的下游发生酪氨酸磷酸化。我们还发现,p34在减数分裂Ⅰ和有丝分裂Ⅰ之前经历了酪氨酸磷酸化和去磷酸化的循环,但在第2至12个有丝分裂细胞周期中未检测到其作为磷酸酪氨酸蛋白的存在。可能在缺乏G2期的细胞周期中,不需要p34cdc2酪氨酸磷酸化所提供的细胞周期蛋白 - p34cdc2复合物组装和激活之间的延迟。最后,一种迁移率在100至116 kDa之间的未鉴定蛋白质或一组蛋白质,在整个成熟过程中酪氨酸磷酸化增加,在受精后10分钟内被去磷酸化或降解,并且在早期胚胎发育过程中似乎在低分子量形式和高分子量形式之间循环。