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Functional analysis of a novel gene, Oogenesin, which localizes to the nucleus at the time of ZGA

ZGA 时定位于细胞核的新基因 Oogenesin 的功能分析

基本信息

批准号：
16380187
负责人：
MINAMI Naojiro
金额：
$ 3.9万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16380187/
关键词：
Oog1 oocyte-specific zygotic gene activation nuclear localization RalGDS Ran Importin β Chchd6 Yeast Two-Hybrid Rasシグナル

项目摘要

In the present study, we performed functional analysis of oocyte-specific gene, Oogenesin (Oog1). We recently demonstrated that Oog1 translocates to the nucleus at the late 1-cell to early 2-cell stage in mouse embryos. The periods coincide with the time of the first gene expression during early preimplantation development that has been termed 'zygotic gene activation (ZGA).' The time of ZGA depends on species ; however, the precise mechanism of ZGA remains unknown. We speculate that Oog1 has some role in ZGA because Oog1 disappears suddenly after ZGA. First of all, to identify the interacting protein with Oog1, we performed yeast two-hybrid experiment. As a result, we identified RalGDS and Chchd6 protein that interact with Oog1. These proteins are also maternally derived products and express specifically in oocyte and preimplantation embryos during early development. Since RalGDS is one of the Ras signaling proteins, Oog1 may function under stream of Ras signaling. RalGDS and Chchd6 have nuclear localization signal suggesting that nuclear localization of Oog1 depend on these proteins. A novel protein Chchd6 has CHCH domain that is essential for the interaction with Oog1. The expression pattern of RalGDS and Chchd6 is quite similar with that of Oog1, suggesting the function of these protein in early embryogenesis. In addition, Oog1 interact with Importin b and Ran in GTP dependent manner, suggesting the involvement of G-protein in Oog1 function.

本研究对卵母细胞特异性基因Oog 1进行了功能分析。我们最近证明，Oog 1在小鼠胚胎1细胞晚期至2细胞早期转移到细胞核。这一时期与早期胚胎植入前发育过程中第一次基因表达的时间相吻合，这一时期被称为合子基因激活（ZGA）。ZGA的时间取决于物种;然而，ZGA的确切机制仍然未知。我们推测Oog 1在ZGA中有一定的作用，因为Oog 1在ZGA后突然消失。首先，我们进行了酵母双杂交实验，鉴定了与Oog 1相互作用的蛋白。结果，我们鉴定了与Oog 1相互作用的RalGDS和Chchd 6蛋白。这些蛋白质也是母体衍生的产物，并且在早期发育过程中特异性地在卵母细胞和植入前胚胎中表达。由于RalGDS是Ras信号传导蛋白之一，因此Oog 1可能在Ras信号传导流下起作用。RalGDS和Chchd 6具有核定位信号，表明Oog 1的核定位依赖于这些蛋白。一种新的蛋白质Chchd 6具有CHCH结构域，该结构域对于与Oog 1的相互作用是必需的。RalGDS和Chchd 6的表达模式与Oog 1的表达模式非常相似，提示这些蛋白在早期胚胎发生中的功能。此外，Oog 1与Importin B和Ran以GTP依赖的方式相互作用，提示G蛋白参与了Oog 1的功能。