Tachibana K, Machida T, Nomura Y, Kishimoto T
Laboratory of Cell and Developmental Biology, Faculty of Biosciences and Biotechnology, Tokyo Institute of Technology, Midoriku, Yokohama, Japan.
EMBO J. 1997 Jul 16;16(14):4333-9. doi: 10.1093/emboj/16.14.4333.
The mechanism by which fertilization initiates S-phase in the zygote is examined by manipulating the activity of MAP kinase in mature starfish eggs. These unfertilized eggs, which are arrested at G1-phase after the completion of meiosis, have high MAP kinase activity but undetectable cdc2 kinase activity. Either fertilization or inhibition of protein synthesis causes a decrease in MAP kinase activity, which is followed by DNA synthesis. Inactivation of MAP kinase with its specific phosphatase, CL100, initiates DNA synthesis in the absence of fertilization, while constitutive activation of MAP kinase with MEK represses the initiation of DNA synthesis following fertilization. Thus, in unfertilized mature starfish eggs, a capacity for DNA replication is already acquired, but entry into S-phase is negatively regulated by MAP kinase activity that is supported by a continuously synthesized protein(s) but not by cdc2 kinase. Upon fertilization, downregulation of MAP kinase activity is necessary and sufficient for triggering the G1/S-phase transition.
通过操纵成熟海星卵中丝裂原活化蛋白激酶(MAP激酶)的活性,研究了受精引发合子进入S期的机制。这些未受精的卵在减数分裂完成后停滞于G1期,具有高MAP激酶活性,但检测不到细胞周期蛋白依赖性激酶2(cdc2激酶)活性。受精或抑制蛋白质合成都会导致MAP激酶活性降低,随后进行DNA合成。用其特异性磷酸酶CL100使MAP激酶失活,在未受精的情况下启动DNA合成,而用丝裂原活化蛋白激酶激酶(MEK)组成性激活MAP激酶则会抑制受精后DNA合成的起始。因此,在未受精的成熟海星卵中,已经具备了DNA复制的能力,但进入S期受到MAP激酶活性的负调控,这种活性由持续合成的一种或多种蛋白质维持,而非由cdc2激酶维持。受精后,MAP激酶活性的下调对于触发G1/S期转换是必要且充分的。