Tudor Domain Proteins in Germline Genome Defense

种系基因组防御中的 Tudor 结构域蛋白

• 批准号: 10394251
• 负责人: Chen Chen
• 金额: $ 32.28万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10394251
• 关键词: Affinity; Animals; Area; Bacteria; Binding; Binding Proteins; Biochemical; Biogenesis; Biology; Cells; DNA Damage; DNA Transposable Elements; Data; Defense Mechanisms; Development; Developmental Biology; Drosophila genus; Family; Fertility; G-Quartets; Gametogenesis; Genetic Transcription; Genome; Genomics; Germ Cells; Immune system; Individual; Infertility; Knowledge; Light; Male Infertility; Meiosis; Mus; Mutation; N-terminal; Parasites; Pathway interactions; Phase; Plants; Play; Production; Property; Proteins; RNA; RNA Binding; RNA Helicase; RNA Precursors; RNA Processing; RNA-Binding Proteins; RNA-Protein Interaction; Regulation; Reporting; Research; Role; Small RNA; Spermatocytes; Spermatogenesis; Structure; Subgroup; Tertiary Protein Structure; Testing; Transcriptional Silencer Elements; base; fetal; genetic information; genome integrity; helicase; insight; male fertility; mouse model; next generation; novel; sperm cell

PROJECT SUMMARY Transposable elements (TEs) are “genomic parasites” that can replicate and re-integrate into the host cell genome. Uncontrolled TE activity in germ cells leads to DNA damage, disruption of gametogenesis, and infertility. In mammalian male germ cells, the PIWI- piRNA pathway uses small RNAs as a guide to silence mobile TEs to protect genome integrity and sustain fertility. The proper production of piRNAs is critical for TE silencing and spermatogenesis. However, the mechanisms governing piRNA biogenesis are not well understood. In particular, the roles of many RNA binding proteins during piRNA biogenesis remain elusive. By studying a subgroup of Tudor domain proteins that also harbor LOTUS domains, we discovered that TDRD5 is a novel RNA binding protein critical for piRNA biogenesis in mice. Strikingly, we have discovered a novel RNA binding property of LOTUS domains that is conserved in bacteria, plants and animals. This binding property is different from reported protein binding feature of some animal LOTUS domains. We hypothesize that animal LOTUS domains have both RNA and protein binding activities and that the LOTUS group of Tudor domain proteins together play critical roles in mammalian piRNA biogenesis. To test this hypothesis, we will use biochemical approaches and mouse models to: 1) Clarify the RNA and protein binding activities of the LOTUS domain superfamily; 2) Determine the mechanism of a specific LOTUS domain-RNA interaction; and 3) Define the functional involvement of LOTUS domain proteins in piRNA biogenesis and Vasa regulation in mice. These studies will provide valuable new insights into the mechanisms of piRNA biogenesis that safeguard germline genome integrity to sustain male fertility and expand our knowledge in broader RNA biology and developmental biology.

项目概要 转座元件（TE）是可以复制和重新整合的“基因组寄生虫” 进入宿主细胞基因组。生殖细胞中不受控制的 TE 活性会导致 DNA 损伤， 配子发生破坏和不育。在哺乳动物雄性生殖细胞中，PIWI- piRNA 途径使用小 RNA 作为指导来沉默移动 TE 以保护基因组 完整性和维持生育能力。 piRNA 的正确生产对于 TE 沉默至关重要 和精子发生。然而，piRNA 生物合成的调控机制并不明确。 很好理解。特别是，许多 RNA 结合蛋白在 piRNA 过程中的作用 生物起源仍然难以捉摸。通过研究 Tudor 结构域蛋白的一个亚组， 含有LOTUS结构域，我们发现TDRD5是一种新型RNA结合蛋白 对小鼠体内的 piRNA 生物合成至关重要。令人惊讶的是，我们发现了一种新的RNA LOTUS 结构域的结合特性在细菌、植物和动物中是保守的。 这种结合特性与报道的某些动物的蛋白质结合特性不同 LOTUS 域。我们假设动物 LOTUS 结构域同时具有 RNA 和 蛋白质结合活性与 Tudor 结构域蛋白质的 LOTUS 组结合在一起 在哺乳动物 piRNA 生物发生中发挥关键作用。为了检验这个假设，我们将使用 生化方法和小鼠模型：1) 阐明 RNA 和蛋白质结合 LOTUS 域超家族的活动； 2）确定具体的机制 LOTUS 结构域-RNA 相互作用； 3) 定义 LOTUS 的功能参与 小鼠 piRNA 生物合成和 Vasa 调节中的结构域蛋白。这些研究将 为 piRNA 生物发生机制提供有价值的新见解，从而保障 种系基因组完整性可维持男性生育能力并扩大我们在更广泛领域的知识 RNA生物学和发育生物学。