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在日本林蛙卵母细胞成熟过程中,细胞周期蛋白B1或B2对于诱导生发泡破裂是必需且充分的。

Either cyclin B1 or B2 is necessary and sufficient for inducing germinal vesicle breakdown during frog (Rana japonica) oocyte maturation.

作者信息

Ihara J, Yoshida N, Tanaka T, Mita K, Yamashita M

机构信息

Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo, Japan.

出版信息

Mol Reprod Dev. 1998 Aug;50(4):499-509. doi: 10.1002/(SICI)1098-2795(199808)50:4<499::AID-MRD14>3.0.CO;2-Y.

Abstract

Oocyte maturation is finally triggered by the maturation-promoting factor (MPF), which consists of Cdc2 and cyclin B. We have cloned cDNAs encoding frog (Rana japonica) cyclins B1 and B2 and produced antibodies against their products. Using the antibodies, we investigated changes in protein states and levels of Cdc2 and cyclins B1 and B2 during oocyte maturation. In immature oocytes, all Cdc2 was a monomeric unphosphorylated inactive 35 kDa form and neither cyclin B1 nor cyclin B2 was present. Mature oocytes contained the MPF complex consisting of an active 34 kDa Cdc2 phosphorylated on threonine161 and a 49 kDa cyclin B1 or a 51 kDa cyclin B2. After progesterone stimulation, both cyclins B1 and B2 were synthesized from their stored mRNAs and bound to the preexisting 35 kDa Cdc2. The binding of Cdc2 with cyclin B and its activation probably through the phosphorylation on threonine161 occurred at almost the same time, in accordance with an electrophoretic mobility shift of Cdc2 from 35 to 34 kDa. Microinjection into immature oocytes of cyclin B1 or B2 mRNA alone, or a mixture of them, induced germinal vesicle breakdown (GVBD) with similar dose-dependence. When the translation of endogenous mRNAs of both cyclins B1 and B2 was inhibited with antisense RNAs, progesterone failed to induce GVBD in the oocytes, but the inhibition of only one of the two was unable to inhibit the progesterone-induced GVBD. These results indicate that either cyclin B1 or B2 is necessary and sufficient for inducing GVBD during Rana oocyte maturation.

摘要

卵母细胞成熟最终由成熟促进因子(MPF)触发,MPF由细胞周期蛋白依赖性激酶2(Cdc2)和细胞周期蛋白B组成。我们克隆了编码青蛙(日本林蛙)细胞周期蛋白B1和B2的cDNA,并制备了针对其产物的抗体。利用这些抗体,我们研究了卵母细胞成熟过程中Cdc2以及细胞周期蛋白B1和B2的蛋白质状态和水平的变化。在未成熟卵母细胞中,所有Cdc2均为单体形式的未磷酸化无活性35 kDa蛋白,细胞周期蛋白B1和B2均不存在。成熟卵母细胞含有由苏氨酸161位点磷酸化的活性34 kDa Cdc2和49 kDa细胞周期蛋白B1或51 kDa细胞周期蛋白B2组成的MPF复合物。孕酮刺激后,细胞周期蛋白B1和B2均从储存的mRNA中合成,并与预先存在的35 kDa Cdc2结合。Cdc2与细胞周期蛋白B的结合及其通过苏氨酸161位点磷酸化的激活几乎同时发生,这与Cdc2的电泳迁移率从35 kDa变为34 kDa一致。单独向未成熟卵母细胞显微注射细胞周期蛋白B1或B2的mRNA,或它们的混合物,会以相似剂量依赖性诱导生发泡破裂(GVBD)。当用反义RNA抑制细胞周期蛋白B1和B2的内源性mRNA翻译时,孕酮无法诱导卵母细胞发生GVBD,但仅抑制其中之一则无法抑制孕酮诱导的GVBD。这些结果表明,在日本林蛙卵母细胞成熟过程中,细胞周期蛋白B1或B2对于诱导GVBD是必要且充分的。

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