Investigating how the conserved ZNFX-1 protein regulates epigenetic inheritance and germline immortality in Caenorhabditis elegans

研究保守的 ZNFX-1 蛋白如何调节秀丽隐杆线虫的表观遗传和种系永生

• 批准号: 10371223
• 负责人: Daniel Joseph Durning
• 金额: $ 3.17万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10371223
• 关键词: Affect; Animals; Binding; Binding Sites; Biochemical; Biochemistry; Biogenesis; C-terminal; Caenorhabditis elegans; Cell Survival; Chromatin Remodeling Factor; Computational Biology; Cysteine; Cytoplasmic Granules; DNA Damage; DNA Transposable Elements; Data; Development; Epigenetic Process; Equilibrium; Family; Fertility; Future; Gene Expression; Generations; Genes; Genetic; Germ Cells; Goals; Guide RNA; Health; Heritability; Measures; Mutation; Nuclear Pore; Pathway interactions; Pattern; Peptides; Phenotype; Population; Positioning Attribute; Postdoctoral Fellow; Proliferating; Proteins; RNA; RNA Binding; RNA-Directed RNA Polymerase; Reporter; Research; Research Personnel; Signal Transduction; Site; Small RNA; Sterility; System; Systems Biology; Temperature; Testing; Training; Transcript; Transgenes; Zinc Fingers; career; epigenetic silencing; epigenome; genome integrity; helicase; mortality; mutant; null mutation; programs; recruit; transcriptome

Project Summary Germ cell immortality is essential for fertility and for species survival. To proliferate indefinitely, germ cells depend on mechanisms that maintain genome integrity and epigenetic programs. In animals, small RNAs that interact with Argonaute proteins of the PIWI family—called piRNAs—serve as a vanguard of transcriptome and epigenome integrity in the germline, by identifying and silencing transposable elements and by regulating germline gene expression. In many animals, piRNAs are essential for germline health. Defective piRNA biogenesis or function activates transposon expression and mobility, increases DNA damage, disrupts germ cell development, and reduces fertility. In Caenorhabditis elegans, small RNA pathways with opposing activities collaborate to maintain germ cell survival and fertility. Recent studies identified ZNFX-1 as a regulator of epigenetic inheritance in worms. ZNFX- 1 is a highly conserved UPF1-like helicase with C-terminal NF-X1-type zinc finger domains. znfx-1 mutants activate epigenetically silenced reporters, and in some cases they silence normally active reporters, indicating that ZNFX-1 balances opposing epigenetic programs. The targeting pattern of small RNAs redistributes in znfx- 1 mutants, suggesting that ZNFX-1 determines the origin of small RNAs required for silencing and anti-silencing pathways. Preliminary data also show that znfx-1 null mutations cause a mortal germline phenotype at elevated temperatures, suggesting that ZNFX-1 maintains balanced epigenetic signals essential for germline immortality. This proposal seeks to use genetic, computational, and biochemical approaches to test the hypothesis that ZNFX-1 is recruited to targets and identifies sites of small RNA biogenesis. Studies in Aim 1 will determine how ZNFX-1 regulates small RNA biogenesis by identifying how and where it binds to target transcripts, and if it unwinds or moves along RNA. Studies in Aim 2 will determine how ZNFX-1 regulates germline immortality by identifying functional domains of ZNFX-1, potentially redundant proteins with homologous domains, and small RNA and transcriptome features of germline immortality. These studies will reveal how animals transmit heritable epigenetic information and how epigenetic pathways maintain germ cell immortality. In addition, the proposed research will provide training in genetics and epigenetics, quantitative biochemistry, and computational approaches, and prepare the fellow for a postdoc in computational and systems biology and a future career as an independent investigator.

项目摘要 生殖细胞的永生是繁殖和物种生存所必需的。为了无限增殖，生殖细胞 关于维持基因组完整性和表观遗传程序的机制。在动物中， PIWI家族的Argonaute蛋白-称为piRNA-作为转录组的先锋， 通过鉴定和沉默转座因子以及通过调节 生殖系基因表达在许多动物中，piRNA对生殖系健康至关重要。缺陷皮尔纳 生物发生或功能激活转座子表达和移动性，增加DNA损伤，破坏生殖细胞 发展，降低生育率。 在秀丽隐杆线虫中，具有相反活性的小RNA通路合作维持生殖细胞 生存和生育能力。最近的研究确定ZNFX-1作为蠕虫表观遗传的调节因子。公司简介 1是一个高度保守的UPF 1样解旋酶，具有C-末端NF-X1型锌指结构域。znfx-1突变体 激活表观遗传沉默的记者，在某些情况下，他们沉默正常活跃的记者，表明 ZNFX-1平衡了对立的表观遗传程序小RNA的靶向模式在znfx中重新分布， 1突变体，表明ZNFX-1决定了沉默和抗沉默所需的小RNA的来源。 途径。初步数据还表明，znfx-1无效突变导致一个致命的生殖细胞表型， 这表明ZNFX-1保持了平衡的表观遗传信号，这对种系永生至关重要。 该提案试图使用遗传、计算和生物化学方法来检验以下假设： ZNFX-1被募集到靶点并识别小RNA生物合成的位点。目标1中的研究将确定如何 ZNFX-1通过识别它如何以及在哪里与靶转录物结合来调节小RNA的生物合成， 沿着沿着RNA展开或移动。目标2中的研究将确定ZNFX-1如何通过以下方式调节生殖系永生： 鉴定ZNFX-1的功能结构域，具有同源结构域的潜在冗余蛋白，以及小的 生殖细胞永生的RNA和转录组特征。这些研究将揭示动物是如何将遗传的 表观遗传信息以及表观遗传途径如何维持生殖细胞的永生。此外，拟议的 研究将提供遗传学和表观遗传学，定量生物化学和计算 方法，并准备在计算和系统生物学博士后和未来的职业生涯， 独立调查员