Kobayashi H, Minshull J, Ford C, Golsteyn R, Poon R, Hunt T
Imperial Cancer Research Fund Clare Hall Laboratories, South Mimms, Potters Bar, Herts, England.
J Cell Biol. 1991 Aug;114(4):755-65. doi: 10.1083/jcb.114.4.755.
We have measured the levels of cyclin mRNAs and polypeptides during oogenesis, progesterone-induced oocyte maturation, and immediately after egg activation in the frog, Xenopus laevis. The mRNA for each cyclin is present at a constant level of approximately 5 x 10(7) molecules per oocyte from the earliest stages of oogenesis until after fertilization. The levels of polypeptides show more complex patterns of accumulation. The B-type cyclins are first detectable in stage IV and V oocytes. Cyclin B2 polypeptide is present at approximately 2 x 10(9) molecules (150 pg) per oocyte by stage VI. The amount increases after progesterone treatment, but returns to its previous level after GVBD and undergoes no further change until it is destroyed at fertilization. Cyclin B1 is present at 4 x 10(8) molecules per oocyte in stage VI oocytes, and rises steadily during maturation, ultimately reaching similar levels to cyclin B2 in unfertilized eggs. Unlike the B-type cyclins, cyclin A is barely detectable in stage VI oocytes, and only starts to be made in significant amounts after oocytes are exposed to progesterone. A portion of all the cyclins are destroyed after germinal vesicle breakdown (GVBD), and cyclins B1 and B2 also experience posttranslational modifications during oocyte maturation. Progesterone strongly stimulates both cyclin and p34cdc2 synthesis in these oocytes, but whereas cyclin synthesis continues in eggs and after fertilization, synthesis of p34cdc2 declines strongly after GVBD. The significance of these results is discussed in terms of the activation and inactivation of maturation-promoting factor.
我们已测定了非洲爪蟾在卵子发生、孕酮诱导的卵母细胞成熟过程以及卵子激活后即刻,细胞周期蛋白mRNA和多肽的水平。从卵子发生的最早阶段直至受精后,每个细胞周期蛋白的mRNA在每个卵母细胞中以大约5×10⁷个分子的恒定水平存在。多肽水平呈现出更为复杂的积累模式。B型细胞周期蛋白最早在IV期和V期卵母细胞中可检测到。到VI期时,每个卵母细胞中细胞周期蛋白B2多肽的含量约为2×10⁹个分子(150皮克)。孕酮处理后该含量增加,但在生发泡破裂(GVBD)后又恢复到先前水平,直至受精时被降解之前不再发生进一步变化。在VI期卵母细胞中,每个卵母细胞中细胞周期蛋白B1的含量为4×10⁸个分子,在成熟过程中稳步上升,最终在未受精卵中达到与细胞周期蛋白B2相似的水平。与B型细胞周期蛋白不同,在VI期卵母细胞中几乎检测不到细胞周期蛋白A,只有在卵母细胞暴露于孕酮后才开始大量合成。所有细胞周期蛋白的一部分在生发泡破裂(GVBD)后被降解,并且细胞周期蛋白B1和B2在卵母细胞成熟过程中也经历翻译后修饰。孕酮强烈刺激这些卵母细胞中细胞周期蛋白和p34cdc2的合成,但是尽管细胞周期蛋白的合成在卵子中以及受精后仍在继续,p34cdc2的合成在GVBD后却大幅下降。我们从成熟促进因子的激活和失活方面讨论了这些结果的意义。