Dynamic regulation of mRNA transport and translation in the Drosophila germline

果蝇种系中 mRNA 运输和翻译的动态调控

• 批准号: 420693198
• 负责人: Dr. Anne Ephrussi, Ph.D.
• 金额: --
• 依托单位: Europäisches Laboratorium für Molekularbiologie (EMBL)
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/420693198?language=en
• 关键词: Dynamic; regulation; mRNA; transport; translation

mRNA localization is a powerful and evolutionarily conserved strategy that coupled to translational control allows precise spatial and temporal control of gene expression within cells. oskar mRNA, whose protein product induces germline formation and posterior patterning in the Drosophila embryo, is a paradigm for mRNA localization and translational control. Key components regulating oskar ribonucleoprotein particle (RNP) behavior include the Exon Junction Complex (EJC) and the associated spliced oskar localization element (SOLE), the double-stranded RNA-binding proteins Egalitarian and Staufen, and importantly dynein and kinesin-1, microtubule motor proteins of opposite polarity that transport oskar mRNA sequentially from its site of synthesis in the nurse cells to the oocyte posterior pole. To gain deeper insight into RNP assembly and regulation, in the first funding period, we developed a stringent biochemical assay, ipaRT, with which we elucidated the Drosophila EJC binding landscape and the factors affecting EJC positioning transcriptome-wide in adult flies, including on oskar mRNA. We also developed a transcript-specific RNP purification method and applied it to oskar under conditions where the mRNA is either transported or sequestered in processing bodies, and thus identified previously known and new candidate oskar mRNP components, including translation regulators. In the second funding period, we will (1) assess the roles of several of these candidate proteins in RNA regulation in vivo and (2) address the roles of the EJC, Egalitarian and Staufen in mRNP assembly and the switch from dynein to kinesin during oskar mRNA transport.

mRNA定位是一种强大而进化的保守策略，耦合到翻译控制的策略可以精确地对细胞内基因表达进行精确的空间和临时控制。 Oskar mRNA的蛋白质产物在果蝇胚胎中诱导种系形成和后型，是mRNA定位和转化控制的范式。 Key components regulating oskar ribonucleoprotein particle (RNP) behavior include the Exon Junction Complex (EJC) and The associated spliced ​​oskar localization element (SOLE), the double-stranded RNA-binding proteins Egalitarian and Staufen, and importantly dynein and kinesin-1, microtubule motor proteins of opposite polarity that transport oskar mRNA sequentially from its site of护士细胞与卵母细胞后极的合成。为了更深入地了解RNP组装和调节，在第一个资金期间，我们开发了一种严格的生化测定，IPART，我们阐明了果蝇EJC结合景观以及影响EJC定位在成年蝇中（包括Oskar mRNA）的EJC定位的因素。我们还开发了一种转录本特异性的RNP纯化方法，并将其应用于OSKAR，在MRNA在处理体中运输或隔离的条件下，因此鉴定出了先前已知的和新的候选OSKAR MRNP组件，包括翻译调节剂。在第二个融资期间，我们将（1）评估这些候选蛋白中的几种在体内RNA调节中的作用，（2）解决EJC，egalitaranian和Staufen在MRNP组装中的作用，以及在OSKAR mRNA运输过程中从Dynein到运动素的转换。