P granules and control of germ cell development

P颗粒和生殖细胞发育的控制

基本信息

批准号：
10005391
负责人：
Ekaterina Voronina
金额：
$ 29.34万
依托单位：
UNIVERSITY OF MONTANA
依托单位国家：
美国
项目类别：
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/10005391
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

项目摘要

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.

抽象的生殖细胞具有多细胞生物的生殖功能和正常生殖细胞开发确保该物种的生存。细菌颗粒是保守的细胞质细胞器细胞，对于这些细胞的生存，分化和功能所必需的细胞。胚芽颗粒成分中的突变或失去其表达导致模型生物的不育症，并且与人类不孕有关。经过对比，体细胞中生殖颗粒成分的不当表达与癌变有关人类。发现许多RNA结合蛋白和发育调节的mRNA富含生殖颗粒，导致假设这些细胞器在调节mRNA稳定性或翻译的调节中起作用活动;然而，这些细胞器的分子功能仍然不确定。线虫C.秀丽隐杆线虫对分析种系中的mRNA平移控制有助于发展。通过专注于促进RNA结合调节蛋白FBF-2的定位的辅助因子到种系干细胞中的胚芽颗粒（称为P颗粒），我们已经产生了一组创新工具将解决P颗粒调节FBF-2活动的关键机制。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如何与辅因子蛋白DLC-1的相互作用对于颗粒定位很重要，会影响翻译抑制由FBF-2施加； 2）定义P颗粒重塑的机制，该机制专门降低了FBF-2 当干细胞从增殖到分化过渡时，减数分裂的发作； 3）确定DLC-1的作用与核心P颗粒成分结合，以调节胚胎P颗粒的稳定性和募集瞬态P颗粒成分，例如FBF-2到成人种系P颗粒。我们的实验系统是准备揭示特定的分子机制，用于介导生殖颗粒与翻译法规。由于细菌颗粒是保守的细胞器，我们的研究重点是保守的调节蛋白，该模型系统中的研究将提供对的重要见解人类不孕。