Localized mRNA translation on the spindle -an essential mechanism for embryonic cell regulation

项目摘要/摘要 将mRNAs定位到胚胎的特定细胞或细胞中的亚细胞区域，能够 空间控制的蛋白质生产。不同浓度的蛋白质导致不同的细胞命运 许多动物在发育过程中的决定、身体模式和细胞功能。尤其是这个， 在胚胎细胞中，局部翻译的机制似乎是必不可少的，这些细胞很大，并经历了 快速的细胞分裂，需要立即输入特定的蛋白质，而不依赖于从 在牢房的其他地方翻译。事实上，有丝分裂纺锤体是一个亚细胞区域，在那里定位 在各种生物体的卵/胚胎中已有mRNAs的报道。……的机制或过程 然而，主轴上的局部平移几乎没有被识别出来。建议的研究重点 在保守的死盒RNA解旋酶Vasa上，该酶定位于每个卵裂球的纺锤体上 胚胎发生。我们之前确定了VASA在一般翻译调节中的功能及其 功能障碍会导致海胆胚胎蛋白质合成减少约80%，并导致有丝分裂缺陷。 因此，我们假设在这些胚胎细胞中，VasA参与了mRNA定位的过程 主轴上的平移。为了验证我们的假设，使用海胆胚胎，这些胚胎表现出恒定的和 在每个卵裂球的纺锤体上都有强大的VASA表达，我们将在体内和实时地可视化 本地化翻译并验证每个mRNA/蛋白质在每个亚细胞中的差异调控方式 区域在胚胎发育过程中。然后我们将通过操纵VASA的亚细胞功能来确定VASA的亚细胞功能 使用先进的成像技术，如光遗传学和生色团，在细胞中进行定位- 辅助光灭活(CALI)。一个本地化的mRNA翻译的生物事件以及一个死亡- 盒RNA解旋酶Vasa在各种生物和细胞中高度保守。这样做的结果是 因此，该项目将对所有研究人员都有用，特别是那些研究亚细胞水平的mrna 蛋白质调节影响任何细胞和生物体的细胞调节和发育。

Project Summary/Abstract Localization of mRNAs to specific cells of an embryo, or to sub-cellular regions in the cell, enables spatially controlled protein production. Distinct concentrations of proteins lead to differential cell fate determination, body patterning, and cell function during development in many animals. Especially, this mechanism of localized translation appears essential in embryonic cells that are large and that undergo rapid cell divisions, requiring immediate input of specific protein without relying on diffusion kinetics from translation elsewhere in the cell. Indeed, the mitotic spindle is a sub-cellular region where localization of mRNAs has been reported in eggs/ embryos of various organisms. The mechanism or process of localized translation on the spindle has been, however, little identified. The proposed research focuses on a conserved DEAD-box RNA helicase Vasa that is localized on the spindle of every blastomere during embryogenesis. We previously identified Vasa functions in general translational regulation and its dysfunction results in ~80% reduction of protein synthesis and mitotic defects in the sea urchin embryo. We, therefore, hypothesize in these embryonic cells, Vasa contributes to the process of localized mRNA translation on the spindle. To test our hypothesis, using sea urchin embryos that show constant and robust Vasa expression on the spindle of every blastomere, we will visualize in vivo and in real time localized translation and validate how each mRNA/protein is differentially regulated at each sub-cellular region during embryogenesis. We will then identify sub-cellular function of Vasa by manipulating its localization in the cell using advanced imaging techniques such as optogenetics and Chromophore- Assisted Light Inactivation (CALI). A biological event of localized mRNA translation as well as a DEAD- box RNA-helicase Vasa is highly conserved among various organisms and cells. The outcomes of this project will be thus useful to all researchers, especially those who study how sub-cellular level of mRNA and protein regulation impacts cellular regulation and development in any cells and organisms.