Regulation of zygotic gene activation in mammals.

哺乳动物合子基因激活的调节。

• 批准号: 10660266
• 负责人: AOKI Fugaku
• 金额: $ 2.3万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10660266/
• 关键词: preimplantation embryo; gene expression; poly(A) tail; 母性mRNA

The aim of this study is to examine the involvement of poly(A) tail elongation of maternal mRNA in the regulation of zygotic gene activation after fertilization. The experiments were conducted that the effect of 3'-deoxiadenosine (3'-dA) on gene activation in mouse 1-cell embryos. After incorporated into the embryos, 3'-dA is expected to be metabolized into 3'-dATP, which inhibits the addition of adenosine to poly(A) tail, working as a chain terminator.The inhibitory effect of 3'-dA on poly(A) tail elongation was confirmed by examining the cyclin A mRNA ; the poly(A) tail of cyclin A mRNA is elongated and its translation starts after fertilization, The results showed that 3'-dA inhibited the elongation of poly(A) tail and translation of cyclin A mRNA. Such an inhibition of translation was specific for cyclin A, since total protein synthesis estimated by [ィイD135ィエD1S] methionine incorporation into macro molecule was not changed by 3'-dA.Embryos were incubated with 3'-dA and the transcriptional activity was measured by in vitro transcription assay 14 h after fertilization. Two mM 3'-dA almost completely inhibited the transcriptional activity. 3'-dG examined as a control did not show any effect.These results suggest that poly(A) tail elongation of maternal mRNA triggers the zygotic gene activation after fertilization.

本研究的目的是探讨母体mRNA的poly(A)尾部延长在受精后合子基因激活的调节中的作用。通过实验观察3'-脱氧腺苷(3'-dA)对基因的影响。小鼠 1 细胞胚胎中的激活后，3'-dA 预计会代谢为 3'-dATP，从而抑制腺苷的添加。通过检测cyclin A mRNA，证实了3'-dA对poly(A)尾延长的抑制作用；cyclin A mRNA的poly(A)尾被延长，其受精后开始翻译，结果表明，3'-dA 抑制聚腺苷酸尾的延长和细胞周期蛋白 A mRNA 的翻译，这种翻译抑制对细胞周期蛋白 A 具有特异性。因为通过[D135D1S]甲硫氨酸掺入大分子估计的总蛋白质合成不会被3'-dA改变。将胚胎与3'-dA一起孵育，并在受精后14小时通过体外转录测定来测量转录活性。 '-dA 几乎完全抑制了作为对照的 3'-dG 的转录活性，但没有显示出任何效果。这些结果表明，poly(A) 尾部。受精后母体 mRNA 的延伸触发合子基因激活。

项目成果

Kim, K-J., Mizoguchi, Y., Kuraishi, T., Ygamaguchi, H., Enami. J., Aoki, F., Imakawa, K. and Sakai, S.: "Effects of wearinga nad suckloing on g-casein and prolactin receptor mRNA levels in the mouse mammary gland during lactation."Anim. Sci. Tech.. 69-8.

金，K-J.，沟口，Y.，仓石，T.，Ygamaguchi，H.，Enami。

Kim, J.-Y.: "Effect of weaning and suckling on γ-casein and prolactin receptor mRNA levels in the mouse mammary gland during lactation"Anim. Sci. Tech. 69. 728-732 (1998)

Kim, J.-Y.：“断奶和哺乳对哺乳期小鼠乳腺中 γ-酪蛋白和催乳素受体 mRNA 水平的影响”Anim.Tech. 69. 728-732 (1998)

Aoki, F., Sakai, S. and Kohmato, K.: "Regulation of flagellar bending by cAMP and Ca2+ in hmaster sperm."Mol. Rreprod. Dev.. 53-1. 77-83 (1999)

Aoki, F.、Sakai, S. 和 Kohmato, K.：“hmaster 精子中 cAMP 和 Ca2 对鞭毛弯曲的调节。”Mol。

Kuraishi, T., Sun, Y., Aoki, F., Imakawa, K. and Sakai, S.: "The lactationg mammary gland changes the poly(A) tail length of casein mRNA depending upon the accumulation and removal of milk."Biochem. J.. (in press).

Kuraishi, T.、Sun, Y.、Aoki, F.、Imakawa, K. 和 Sakai, S.：“哺乳期乳腺根据乳汁的积累和排出改变酪蛋白 mRNA 的聚 (A) 尾长度。

Aoki,F.: "Regulation of flagellar bending by cAMP and Ca^ in hamster sperm"Mol. Reprod, Dev.. 53・1. 77-83 (1999)

青木 F.：“仓鼠精子中 cAMP 和 Ca^<2+> 的鞭毛弯曲调节”Mol Reprod，Dev.. 53・1（1999）。