RUI: Biochemical and genetic analysis of conserved molecular scaffold Tudor complex required for germ cell specification in Drosophila

RUI：果蝇生殖细胞规范所需的保守分子支架 Tudor 复合物的生化和遗传分析

• 批准号: 2130162
• 负责人: Alexey Arkov
• 金额: $ 82.24万
• 依托单位: Murray State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2130162&HistoricalAwards=false
• 关键词: RUI; Biochemical; genetic; analysis; conserved

Cellular organization is critically dependent on the self-assembly of various membraneless organelles inside the cell. However, the principles and molecular mechanisms that drive and regulate the assembly of these organelles and define their functions are poorly understood. This project integrates cutting-edge genetics and molecular quantitative approaches to provide major insights into the mechanisms responsible for the assembly and function of evolutionarily conserved membraneless organelles assembled in germ cells of model organism Drosophila (germ granules). These cells give rise to egg and sperm cells, and therefore, are responsible for continuity of life. This research will be integrated into several cell and genetics undergraduate courses and will directly engage undergraduate and graduate students in quantitative cell and molecular biology training.Germ granules show the properties of soft condensed matter and they often change their morphology during developmental transitions. Recent research identified an evolutionarily conserved germ granule multisubunit complex assembled on the large scaffold Tudor protein. Tudor scaffold has 11 protein-protein interaction modules (Tudor domains) and Tudor complex components are required for germ cell specification. The goal of this project is to provide molecular understanding of how this multisubunit Tudor scaffold complex is assembled using purified components and quantitative biochemical approaches. In addition, using super-resolution microscopy, it will be determined how different components of the complex are assembled into large germ granules in vivo. Furthermore, to understand the function of Tudor complex, enzymatic activities of its components upon the complex assembly will be characterized and the significance of the complex for germ granule formation, morphology and germ cell specification will be determined by mutational approaches.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

细胞组织严重依赖于细胞内各种无膜细胞器的自组装。然而，驱动和调节这些细胞器的组装并定义其功能的原理和分子机制知之甚少。 该项目整合了尖端的遗传学和分子定量方法，为负责模式生物果蝇（生殖颗粒）生殖细胞中进化保守的无膜细胞器的组装和功能的机制提供了重要见解。这些细胞产生卵子和精子细胞，因此，负责生命的延续。这项研究将被整合到几个细胞和遗传学本科课程，并将直接从事定量细胞和分子生物学培训本科生和研究生。胚芽颗粒显示出软凝聚态物质的性质，它们经常在发育过渡期间改变形态。最近的研究确定了一个进化上保守的胚芽颗粒多亚基复合物组装在大支架Tudor蛋白。Tudor支架具有11个蛋白质-蛋白质相互作用模块（Tudor结构域），Tudor复合物组分是生殖细胞特异性所需的。该项目的目标是提供分子理解如何使用纯化的组分和定量生化方法组装这种多亚基Tudor支架复合物。此外，使用超分辨率显微镜，将确定复合物的不同组分如何在体内组装成大的胚芽颗粒。此外，为了了解Tudor复合物的功能，将表征其组分在复合物组装时的酶活性，并通过突变方法确定复合物对生殖颗粒形成、形态和生殖细胞特化的重要性。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。