P granules and control of germ cell development

P颗粒和生殖细胞发育的控制

• 批准号: 10174938
• 负责人: Ekaterina Voronina
• 金额: $ 29.32万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10174938
• 关键词: Address; Adult; Animal Model; Animals; Binding; Binding Sites; Biochemical; Biological Models; Caenorhabditis elegans; Cell physiology; Characteristics; Cytoplasmic Granules; Cytoplasmic Organelle; Development; Embryo; Ensure; Fertility; Functional disorder; Gene Expression Profile; Genes; Genetic; Genetic Translation; Germ; Germ Cells; Human; In Vitro; Infertility; Lead; Link; Mediating; Meiosis; Messenger RNA; Molecular; Mutagenesis; Mutate; Mutation; Nematoda; Organelles; Organism; Peptides; Play; Post-Transcriptional Regulation; Proteins; RNA Binding; RNA-Binding Proteins; Regulation; Regulatory Pathway; Reproduction; Research; Role; Site; Somatic Cell; System; Testing; Translational Regulation; Translational Repression; Work; carcinogenesis; cofactor; genetic analysis; genetic regulatory protein; germline stem cells; imaging approach; in vivo; innovation; insight; mRNA Stability; prevent; public health relevance; recruit; reproductive function; stem cells; tool; translational impact

Abstract Germ cells carry out the reproductive function of multicellular organisms, and normal germ cell development ensures survival of the species. Germ granules are conserved cytoplasmic organelles of germ cells, essential for survival, differentiation, and function of these cells. Mutations in germ granule components or loss of their expression lead to infertility in model organisms and are associated with infertility in humans. By contrast, inappropriate expression of germ granule components in somatic cells is linked to carcinogenesis in humans. Many RNA-binding proteins and developmentally regulated mRNAs are found enriched in germ granules, leading to a hypothesis that these organelles function in regulation of mRNA stability or translational activity; yet, the molecular function of these organelles is still undefined. The nematode C. elegans has been instrumental for analysis of mRNA translational control in germline development. Through focusing on cofactors that promote localization of RNA-binding regulatory protein FBF-2 to germ granules (called P granules in this species) in germline stem cells we have generated a set of innovative tools that will address key mechanisms through which P granules regulate FBF-2 activity. By integrating genetic, molecular, biochemical and imaging-based approaches, we will: 1) Determine how FBF-2 interaction with a cofactor protein DLC-1 important for P granule localization impacts translational repression exerted by FBF-2; 2) Define the mechanisms of P granule remodeling that specifically degrade FBF-2 at the onset of meiosis when stem cells transition from proliferation to differentiation; 3) Determine the role of DLC-1 binding to core P granule components in regulation of stability of embryonic P granules and in recruitment of transient P granule components such as FBF-2 to adult germline P granules. Our experimental system is poised to reveal specific molecular mechanisms mediating the interplay between germ granules and translational regulation. Since germ granules are conserved organelles and our research focuses on conserved regulatory proteins, studies in this model system will provide critical insight into the causes of infertility in humans.

抽象的 生殖细胞执行多细胞生物的生殖功能，正常生殖细胞 发展保证了物种的生存。胚芽颗粒是胚芽保守的细胞质细胞器 细胞，对于这些细胞的生存、分化和功能至关重要。胚芽颗粒成分突变 它们的表达或丧失会导致模式生物不育，并与人类不育有关。经过 相反，体细胞中胚芽颗粒成分的不当表达与癌症的发生有关 人类。细菌中富含许多 RNA 结合蛋白和发育调节 mRNA 颗粒，导致假设这些细胞器在调节 mRNA 稳定性或翻译中起作用 活动;然而，这些细胞器的分子功能仍不清楚。 线虫 C. elegans 有助于分析种系中 mRNA 翻译控制 发展。通过关注促进 RNA 结合调节蛋白 FBF-2 定位的辅助因子 对于生殖系干细胞中的生殖颗粒（在该物种中称为 P 颗粒），我们生成了一组 创新工具将解决 P 颗粒调节 FBF-2 活性的关键机制。经过 整合遗传、分子、生化和基于成像的方法，我们将： 1) 确定 FBF-2 如何 与对 P 颗粒定位很重要的辅助因子蛋白 DLC-1 的相互作用会影响翻译抑制 由FBF-2施加； 2）定义P颗粒重塑的机制，该机制特异性地降解FBF-2 当干细胞从增殖转变为分化时减数分裂开始； 3）确定DLC-1的作用 与核心 P 颗粒成分结合，调节胚胎 P 颗粒的稳定性并招募 瞬时 P 颗粒成分（例如 FBF-2）到成体种系 P 颗粒。我们的实验系统是 准备揭示介导细菌颗粒和细菌颗粒之间相互作用的特定分子机制 翻译监管。由于胚芽颗粒是保守的细胞器，我们的研究重点是 保守的调节蛋白，对该模型系统的研究将为了解其原因提供重要的见解 人类不育症。