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

纺锤体上的局部 mRNA 翻译——胚胎细胞调节的重要机制

• 批准号: 10529553
• 负责人: Mamiko Yajima
• 金额: $ 6.8万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-26 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10529553
• 关键词: Animals; Biological; Body Patterning; Cell division; Cell physiology; Cells; Defect; Development; Diffusion; Embryo; Embryonic Development; Event; Functional disorder; Genetic Translation; Imaging Techniques; Kinetics; Lead; Light; Messenger RNA; Mitotic; Mitotic spindle; Organism; Outcome; Process; Production; Protein Biosynthesis; Proteins; RNA Helicase; Regulation; Reporting; Research; Research Personnel; Sea Urchins; Testing; Time; Translational Regulation; Translations; blastomere structure; cell growth regulation; cellular development; chromophore; egg; embryo cell; embryo stage 2; in vivo; insight; optogenetics

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.

项目概要/摘要 将 mRNA 定位到胚胎的特定细胞或细胞中的亚细胞区域，使得 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 许多动物发育过程中的决定、身体模式和细胞功能。尤其是这个 局部翻译机制对于较大且经历过的胚胎细胞来说似乎是至关重要的 快速细胞分裂，需要立即输入特定蛋白质，而不依赖于扩散动力学 翻译到细胞的其他地方。事实上，有丝分裂纺锤体是一个亚细胞区域，其中 据报道，各种生物体的卵/胚胎中都存在 mRNA。其机制或过程 然而，主轴上的局部平​​移却鲜为人知。拟议的研究重点 位于每个卵裂球纺锤体上的保守 DEAD-box RNA 解旋酶 Vasa 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 功能障碍导致海胆胚胎中蛋白质合成减少约 80% 和有丝分裂缺陷。 因此，我们假设在这些胚胎细胞中，Vasa 有助于本地化 mRNA 的过程 主轴上的平移。为了检验我们的假设，使用表现出恒定和 每个卵裂球纺锤体上都有强大的 Vasa 表达，我们将在体内实时可视化 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节方式 胚胎发生过程中的区域。然后我们将通过操纵 Vasa 的亚细胞功能来识别其 使用先进的成像技术（例如光遗传学和发色团）在细胞中进行定位 辅助光失活（CALI）。局部 mRNA 翻译以及 DEAD- 的生物学事件 Box RNA 解旋酶 Vasa 在各种生物体和细胞中高度保守。这样做的结果 因此，该项目将对所有研究人员有用，尤其是那些研究 mRNA 的亚细胞水​​平如何 蛋白质调节影响任何细胞和生物体的细胞调节和发育。

项目成果

The germline factor DDX4 contributes to the chemoresistance of small cell lung cancer cells.

• DOI: 10.1038/s42003-023-04444-7
• 发表时间: 2023-01-18
• 期刊: COMMUNICATIONS BIOLOGY
• 影响因子: 5.9
• 作者: Noyes, Christopher;Kitajima, Shunsuke;Li, Fengkai;Suita, Yusuke;Miriyala, Saradha;Isaac, Shakson;Ahsan, Nagib;Knelson, Erik;Vajdi, Amir;Tani, Tetsuo;Thai, Tran C.;Xu, Derek;Murai, Junko;Tapinos, Nikos;Takahashi, Chiaki;Barbie, David A.;Yajima, Mamiko
• 通讯作者: Yajima, Mamiko

Light-induced, spatiotemporal control of protein in the developing embryo of the sea urchin.

• DOI: 10.1016/j.ydbio.2021.06.006
• 发表时间: 2021-10
• 期刊: Developmental biology
• 影响因子: 2.7
• 作者: Wavreil FDM;Poon J;Wessel GM;Yajima M
• 通讯作者: Yajima M

Micromere formation and its evolutionary implications in the sea urchin.

• DOI: 10.1016/bs.ctdb.2021.10.008
• 发表时间: 2022
• 期刊: Current topics in developmental biology
• 作者: Emura N;Yajima M
• 通讯作者: Yajima M

Vasa nucleates asymmetric translation along the mitotic spindle during unequal cell divisions.

• DOI: 10.1038/s41467-022-29855-8
• 发表时间: 2022-04-20
• 期刊: Nature communications
• 影响因子: 16.6

CRISPR-Cas9 editing of non-coding genomic loci as a means of controlling gene expression in the sea urchin.

• DOI: 10.1016/j.ydbio.2021.01.003
• 发表时间: 2021-04
• 期刊: Developmental biology
• 影响因子: 2.7
• 作者: Pieplow A;Dastaw M;Sakuma T;Sakamoto N;Yamamoto T;Yajima M;Oulhen N;Wessel GM
• 通讯作者: Wessel GM