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Molecular Mechanisms of the Inititaiton of Protein Translation during Oocyte Maturation

卵母细胞成熟过程中蛋白质翻译启动的分子机制

基本信息

批准号：
09680716
负责人：
YAMASHITA Masakane
金额：
$ 2.11万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680716/
关键词：
oocyte maturation protein translation MRF cyclin B mRNA

项目摘要

This study was undertaken to investigate molecular mechanisms of the initiation of cyclin BETA mRNA translation during oocyte maturation, with special reference to the following three subjects.1) Significance of cyclin BETA translation during oocyte maturationOur previous finding that cyclin BETA mRNA translation is necessary for initiating goldfish oocyte maturation has been also confirmed for other species, including fishes other than goldfish and amphibians (frog, toad and newt) except for Xenopus laevis, suggesting the general adoption of this mechanism in lower vertebrates.2) Identification of cyclin BETA mRNA-binding proteins and the effect of hormone on translation.An attempt to identify cyclin BETA mRNA-binding protein purified with a biotin-labeled RNA affinity chromatography was unsuccessful because of the poor yield. To examine the effect of hormonal stimulation on the translational control, we injected cyclin BETA mRNA into honnone-treated oocytes. We found that the progression of oocyte maturation induced by the injection of cyclin BETA mRNA occurred earlier than that induced by the hormone, but later than that induced by the injection cyclin BETA protein. These results indicate that the hormonal treatment enhances the synthesis of cyclin BETA protein from the injected mRNA.3) Involvement of known proteins in cyclin BETA mRNA translational initiationTo investigate the functions during oocyte maturation of proteins that have been already suggested to be translational regulators in other systems, we have isolated cDNA clones and produced antibodies against their products. Those include cytoplasmic polyadenylation element-binding-protein (CPEB), Nanos, Pumilio, poly(A) polymerase, poly(A) binding proteins, Y-box proteins. We have shown that Y-box proteins and Pumilio are a cyclin B mRNA-binding protein, suggesting the involvement of these proteins in the translational control during oocyte maturation.

本研究旨在探讨卵母细胞成熟过程中cyclin BETA mRNA翻译启动的分子机制，主要涉及以下三个方面：1）cyclin BETA mRNA翻译在卵母细胞成熟过程中的意义我们以前的研究发现，cyclin BETA mRNA翻译是启动金鱼卵母细胞成熟所必需的，这一发现也在其他物种中得到了证实，包括金鱼以外的鱼类和两栖类（蛙、蟾蜍和蝾螈），表明该机制在低等脊椎动物中普遍采用。2）细胞周期蛋白BETA mRNA结合蛋白的鉴定及激素对翻译的影响。为了研究激素刺激对翻译控制的影响，我们将细胞周期蛋白BETA mRNA注射到honone处理的卵母细胞中。我们发现，注射cyclin BETA mRNA诱导的卵母细胞成熟进程早于激素诱导的成熟进程，但晚于注射cyclin BETA蛋白诱导的成熟进程。这些结果表明激素处理增强了从注射的mRNA合成细胞周期蛋白BETA蛋白。3）已知蛋白参与细胞周期蛋白BETA mRNA翻译起始为了研究已经被认为是其他系统中的翻译调节因子的蛋白在卵母细胞成熟过程中的功能，我们分离了cDNA克隆并制备了针对其产物的抗体。这些包括细胞质聚腺苷酸化元件结合蛋白（CPEB），Nanos，Pumilio，poly（A）聚合酶，poly（A）结合蛋白，Y盒蛋白。我们已经表明，Y盒蛋白和Pumilio是一个细胞周期蛋白B mRNA结合蛋白，这表明这些蛋白质在卵母细胞成熟过程中的翻译控制的参与。