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.

细胞组织关键取决于细胞内部各种无膜细胞器的自组装。但是，对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。 该项目整合了最先进的遗传学和分子定量方法，以提供有关负责在模型有机体果蝇（胚芽颗粒）生殖细胞中组装进化保守的膜无细胞组装和功能的机制的主要见解。这些细胞会产生卵和精子细胞，因此导致生命的连续性。这项研究将整合到几个细胞和遗传学本科课程中，并将直接参与定量细胞和分子生物学培训的本科生和研究生。Germ颗粒显示了软凝结物质的特性，它们经常在发育过渡期间改变其形态。最近的研究确定了一种在大型脚手铎式蛋白质上组装的进化保守的生殖颗粒多亚基复合物。都铎支架具有11个蛋白质 - 蛋白质相互作用模块（都铎式结构域），而生殖细胞规格需要都铎式复合物成分。该项目的目的是提供分子理解这种多生tudor支架复合物如何使用纯化的组件和定量生化方法组装。另外，使用超分辨率显微镜，将确定如何将复合物的不同成分组装成体内大的胚芽颗粒中。此外，为了了解其成分在复杂组装上的酶促活性的功能，将会表征其组件的酶促活性，并且该复合物对生殖颗粒形成，形态和生殖细胞规格的意义将由突变方法确定。该奖项奖通过评估NSF的法定任务，并通过评估了基金会的智力效果，并反映了与基金会的构成依据。