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.

摘要 生殖细胞执行多细胞生物的生殖功能，正常生殖细胞 人类的发展确保了物种的生存。生殖颗粒是生殖细胞中保守的细胞质细胞器 细胞，对这些细胞的存活、分化和功能至关重要。胚粒突变 在模式生物中，它们的表达的缺失导致不育，并且与模式生物中的不育相关。 人类相比之下，体细胞中胚粒成分的不适当表达与 致癌作用许多RNA结合蛋白和发育调控的mRNA被发现 在胚芽颗粒中富集，导致这些细胞器在mRNA的调节中起作用的假设 稳定性或翻译活性;然而，这些细胞器的分子功能仍然不确定。 线虫C. elegans对于理解生殖细胞的翻译调控是很有帮助的 发展我们以前的研究证明了C.线虫生殖颗粒（P 颗粒）对生殖系干细胞中RNA结合调节蛋白FBF-2的调节作用， 关于这一贡献的机制基础，仍有许多有待了解的地方。我们的实验系统是 准备从分子层面来解决这个问题。我们的研究将集中在FBF-2作为一个范例的细菌 颗粒的贡献，调节生殖细胞中的RNA结合蛋白的活性。通过整合生物化学， 通过分子、遗传和成像技术，我们将：1）确定FBF-2的具体组成部分 2）鉴定FBF-2的辅因子，其依赖于P颗粒完整性用于它们的组装， 3）确定FBF-2介导P颗粒募集和FBF-2特异性调节P颗粒募集的序列。 活动这些研究将揭示胚芽颗粒依赖性调节的一般机制。自从细菌 颗粒是保守的细胞器，FBF-2是保守的PUF蛋白家族的成员， 该模型系统将为人类不孕症的原因提供重要的见解。