RNA binding protein networks and translational control in mammalian oocytes

哺乳动物卵母细胞中的RNA结合蛋白网络和翻译控制

• 批准号: 10388166
• 负责人: Marco Conti
• 金额: $ 34.04万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-06 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10388166
• 关键词: 3' Untranslated Regions; Binding; Biochemical; Biological Process; CDC2 gene; Cell Cycle; Cell Cycle Regulation; Cell Differentiation process; Cell division; Cell physiology; Cells; Chromosome abnormality; Code; Competence; Complex; Data; Data Set; Development; Drosophila genus; Elements; Embryo; Embryonic Development; Epigenetic Process; Event; Experimental Models; Gametogenesis; Gene Expression; Gene Expression Regulation; Genetic Transcription; Genetic Translation; Germ Cells; Growth; Human Pathology; In Vitro; Light; Link; Longevity; Maternal Messenger RNA; Meiosis; Messenger RNA; MicroRNAs; Modeling; Molecular; Monitor; Mus; Normal Cell; Nuclear Envelope; Oocytes; Outcome Study; Pattern; Physiological; Play; Post-Transcriptional Regulation; Process; Property; Proteins; RNA-Binding Proteins; Rana; Regulation; Repression; Ribosomes; Role; Somatic Cell; Testing; Time; Totipotency; Totipotent; Transcriptional Regulation; Translating; Translational Regulation; Translational Repression; Translations; Zebrafish; carcinogenesis; cis acting element; cyclin B1; cyclin B2; experimental study; gastrulation; genome-wide; gonad development; human disease; in vivo; insight; loss of function; mRNA Stability; novel; novel strategies; nuclear reprogramming; oocyte maturation; programs; protein complex; tool; translation assay; zygote

Throughout the lifespan of a cell, a combination of transcriptional and post-transcriptional regulations control gene expression, particularly at the time of critical developmental decisions and during the cell cycle. This is especially true for the development of gametes. Indeed, many pivotal transitions required for the differentiation of these cells rely more on translational regulation of mRNA than on transcription. To overcome the technical hurdles inherent in the germ cell experimental model, we have developed novel strategies to investigate how translation is regulated in mouse oocytes. Monitoring the association of maternal mRNAs with ribosomes together with single oocyte translation assays has allowed us to view the translational landscape in oocytes during growth and maturation in ways previously not possible. By monitoring translation at a genome-wide level, we have identified a global switch in translation that occurs at the time of oocyte reentry into the meiotic cell cycle. Translation of mRNAs required for oocyte growth ceases, while translation of mRNA coding for cell cycle components is activated together with translation of components that will be necessary for embryo development. Using the translation of the critical cell cycle components Cyclin B1 and Cyclin B2 as a paradigm, we have demonstrated that translation defines some unique properties of the meiotic cell cycle. Here, we propose to extend these observations by exploring further how differential translation of maternal mRNAs is regulated during oocyte growth and maturation. The experiments are organized along three specific aims. With the first Specific Aim, we will investigate the intrinsic features of the 3'UTR of mRNAs translated at high levels during oocyte growth and quiescence. We will use novel strategies to identify functional elements in the 3' UTR of an mRNA including those present in the Cyclin B2 mRNA, and to understand how they contribute to translational control in vitro and in vivo. With the second Specific Aim, we will explore the biochemical properties of protein complexes that function as repressors of translation, including those involved in repression of CyclinB1 translation. In the last Specific Aim, we will investigate how repression of translation is triggered at the time of oocyte reentry into the cell cycle. We will establish links between translational repression and regulation of cell cycle components. In addition, we will explore the 3' UTR code that defines timely repression of translation and destabilization. The major outcome of these studies will be a better understanding of the mechanism of repression and activation of maternal mRNA during gamete development. These studies are significant because they will shed light on how mRNA translation is intimately involved in regulation of the cell cycle and how translational programs direct development.

在细胞的整个生命周期中，转录和转录后调控的组合控制 基因表达，特别是在关键发育决策时和细胞周期期间。这是 对于配子的发育尤其如此。事实上，差异化需要许多关键转变 这些细胞更多地依赖于 mRNA 的翻译调节而不是转录。为克服技术难题 由于生殖细胞实验模型固有的障碍，我们开发了新的策略来研究如何 翻译在小鼠卵母细胞中受到调节。监测母体 mRNA 与核糖体的关联 与单卵母细胞翻译测定一起使我们能够观察卵母细胞的翻译景观 以以前不可能的方式生长和成熟。通过在全基因组水平上监测翻译， 我们已经确定了在卵母细胞重新进入减数分裂细胞时发生的全局翻译转换 循环。卵母细胞生长所需的 mRNA 翻译停止，而编码细胞周期的 mRNA 翻译停止 成分与胚胎发育所需成分的翻译一起被激活。 以细胞周期关键成分 Cyclin B1 和 Cyclin B2 的翻译为范例，我们有 证明翻译定义了减数分裂细胞周期的一些独特特性。在此，我们建议 通过进一步探索母体 mRNA 的差异翻译是如何在 卵母细胞的生长和成熟。这些实验是按照三个具体目标组织的。随着第一个具体 目的，我们将研究卵母细胞过程中高水平翻译的 mRNA 3'UTR 的内在特征 生长和静止。我们将使用新的策略来识别 mRNA 3' UTR 中的功能元件 包括 Cyclin B2 mRNA 中存在的那些，并了解它们如何有助于翻译控制 体外和体内。通过第二个具体目标，我们将探索蛋白质复合物的生化特性 发挥翻译抑制因子的作用，包括那些参与抑制 CyclinB1 翻译的因子。在 最后一个具体目标，我们将研究在卵母细胞重新进入卵母细胞时如何触发翻译抑制 细胞周期。我们将建立翻译抑制和细胞周期成分调节之间的联系。 此外，我们将探索定义及时抑制翻译和不稳定的 3' UTR 代码。这 这些研究的主要成果将是更好地理解抑制和激活的机制。 配子发育过程中的母体 mRNA。这些研究意义重大，因为它们将揭示如何 mRNA 翻译与细胞周期的调节以及翻译程序如何指导密切相关 发展。