P granules and control of germ cells development

P颗粒和生殖细胞发育的控制

• 批准号: 8613908
• 负责人: Ekaterina Voronina
• 金额: $ 26.03万
• 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 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8613908
• 关键词: Address; Amino Acid Sequence; Animal Model; Basic Science; Binding; Biochemical; Biological Models; Caenorhabditis elegans; Cell Maintenance; Cell physiology; Cells; Complex; Cytoplasmic Granules; Cytoplasmic Organelle; Development; Ectopic Expression; Ensure; Family Study; Genes; Genetic; Genetic Complementation Test; Germ; Germ Cells; Human; Image; Immunoglobulin Variable Region; Infertility; Lead; Learning; Link; Mediating; Membrane Proteins; Messenger RNA; Modeling; Molecular; Molecular Analysis; Molecular Genetics; Mutation; Nematoda; Organelles; Organism; Peptide Sequence Determination; Post-Transcriptional Regulation; Process; Production; Protein Family; Proteins; Proteomics; RNA; RNA Binding; RNA Interference; RNA-Binding Proteins; Regulation; Regulatory Pathway; Repression; Role; Set protein; Somatic Cell; Specific qualifier value; Stem cells; System; Testing; Transgenes; Transgenic Organisms; Translational Regulation; Translational Repression; Work; base; carcinogenesis; cell type; cofactor; defined contribution; genetic analysis; genetic regulatory protein; insight; mRNA Stability; member; mutant; public health relevance; reproductive function; tumor; tumorigenesis

Abstract Germ cells carry out the reproductive function of the multicellular organisms, and normal germ cell development ensures survival of the species. Germ granules are conserved cytoplasmic organells of the germ cells, essential for the 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 the 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 understanding translational regulation of germline development. Our previous studies demonstrated specific contribution of C. elegans germ granules (P granules) to the regulation exerted by an RNA-binding regulatory protein FBF-2 in germline stem cells, yet much remains to be learned about the mechanistic basics of this contribution. Our experimental system is poised to address this question in molecular detail. Our studies will focus on FBF-2 as a paradigm of germ granule contribution to regulating the activity of RNA-binding proteins in germline. By integrating biochemical, molecular, genetic, and imaging-based approaches, we will: 1) Define the specific components of the FBF-2 regulatory complex; 2) Identify the cofactors of FBF-2 that depend on P granule integrity for their assembly with FBF-2; 3) Determine the sequences of FBF-2 mediating P granule recruitment and FBF-2-specific regulatory activity. These studies will reveal general mechanisms of germ granule-dependent regulation. Since the germ granules are conserved organelles and FBF-2 is a member of conserved PUF protein family, 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，正常生殖细胞 开发确保该物种的生存。生殖颗粒是菌种保守的细胞质细胞器 细胞，对于这些细胞的生存，分化和功能必不可少的细胞。胚芽颗粒中的突变 组件或表达的丧失导致模型生物的不孕症，并且与不孕有关 人类。相比之下，体细胞中生殖颗粒成分的不当表达与 人类的致癌作用。发现许多RNA结合蛋白和发育调节的mRNA 富集在细菌颗粒中，导致假设这些细胞器在MRNA调节中起作用 稳定或翻译活动；然而，这些细胞器的分子功能仍然不确定。 线虫C.秀丽隐杆线虫对理解种系的翻译调节具有重要作用 发展。我们以前的研究表明了秀丽隐杆线虫生殖颗粒的特定贡献（P 颗粒）由RNA结合调节蛋白FBF-2在种系干细胞中施加的调节，但 关于这项贡献的机械基础知识还有很多待汲取的知识。我们的实验系统是 准备通过分子细节解决这个问题。我们的研究将集中于FBF-2作为细菌的范式 颗粒对调节种系中RNA结合蛋白活性的贡献。通过整合生化， 分子，遗传和基于成像的方法，我们将：1）定义FBF-2的特定成分 调节络合物； 2）确定依赖于P颗粒完整性的FBF-2的辅因子的组装 FBF-2; 3）确定FBF-2介导P颗粒募集和FBF-2特异性调节的序列 活动。这些研究将揭示细菌颗粒依赖性调节的一般机制。自从胚芽以来 颗粒是保守的细胞器，FBF-2是保守的PUF蛋白家族的成员。 模型系统将为人类不孕的原因提供批判性的洞察力。