Functions of Piwi proteins and short RNAs in germline

Piwi 蛋白和短 RNA 在种系中的功能

• 批准号: 8046405
• 负责人: ALEXEI A. ARAVIN
• 金额: $ 24.65万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-15 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8046405
• 关键词: Amaze; Animals; Biochemical; Biochemical Genetics; Biogenesis; Biological; Biological Models; Biological Process; Comparative Study; Complex; Cytoplasmic Granules; Defect; Development; Diagnostic; Drosophila genus; Epigenetic Process; Exhibits; Family; Foundations; Gametogenesis; Gene Expression; Gene Silencing; Genes; Genetic Transcription; Genomics; Germ Cells; Human; Infertility; Investigation; Lead; Learning; Mediating; Mentors; Methods; MicroRNAs; Modeling; Molecular; Mus; Organism; Pathway interactions; Phase; Play; Process; Production; Protein Family; Proteins; RNA; RNA Interference; RNA Processing; Reporter; Reporter Genes; Research; Role; Site; Small RNA; Sterility; Structure; System; Testing; Transcript; Transgenic Organisms; Yeasts; in vivo; insight; member; mouse model; mutant; operation; piRNA; programs; therapeutic target; tool

Gene silencing by the RNA interference (RNAi) machinery is an evolutionary conserved process that is critical for control of genes expression in organisms from yeast to human. Targets of RNAi are recognized through complementary interactions with small RNAs that act as guides in the silencing process. Several discrete classes, encompassing thousands of small RNAs have been identified in recent years. For example, microRNAs interact with members of the ubiquitously expressed Argonaute protein family and together these regulate gene expression networks that impact diverse biological processes. Members ofthe other branch of the Argonaute family, the Piwi proteins, are expressed specifically in germ cells with mutant animals showing severe defects in gametogenesis that lead to sterility. Until recently, the small RNA partners of Piwi proteins were unknown, and the molecular functions of Piwis in gametogenesis remain so. We comprehensively characterized Piwi interacting RNA (piRNA) expression in germ cells of mouse and Drosophila. We have also identified the protein partners of Piwi proteins and additional components ofthe cytoplasmic granules that represent the main sites of piRNA pathway operation. In this proposal I present a focused strategy for analyzing piRNA biogenesis and function in the germline. In aim 1,1 will probe the mechanism of piRNA biogenesis in vivo. In aim 2,1 will determine the mechanism of piRNA-Piwi mediated silencing.

通过RNA干扰（RNAi）机械的基因沉默是一种进化保守的过程 从酵母到人的生物中控制基因表达至关重要。 RNAi的靶标被识别 通过与小型RNA的互补相互作用，在沉默过程中充当指南。一些 近年来已经确定了成千上万的小型RNA。例如， MicroRNA与普遍表达的Argonaute蛋白家族的成员相互作用，并将其共同 调节影响各种生物过程的基因表达网络。另一个分支的成员 Argonaute家族，PIWI蛋白，在突变动物的生殖细胞中特别表达 在配子发生中严重缺陷，导致不育。直到最近，Piwi蛋白的小RNA伴侣 是未知的，Piwis在配子发生中的分子功能仍然如此。我们全面 在小鼠和果蝇的生殖细胞中表征了PIWI相互作用的RNA（PIRNA）表达。我们有 还确定了PIWI蛋白的蛋白质伴侣和细胞质颗粒的其他成分 代表了Pirna Pathway操作的主要地点。在此提案中，我提出了一个集中的策略 分析piRNA生物发生和生殖线的功能。在AIM 1,1中将探测piRNA的机制 体内生物发生。在AIM 2,1中，将确定Pirna-Piwi介导的沉默的机理。