RNA binding proteins and mRNA localisation in Drosophila

果蝇中的 RNA 结合蛋白和 mRNA 定位

• 批准号: 2435443
• 金额: --
• 依托单位: CARDIFF UNIVERSITY
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2435443
• 关键词: RNA; binding; proteins; mRNA; localisation

mRNA localisation to specific sub-cellular regions is a widespread phenomenon in polarised cells and is critical both during development (eg for patterning) and in differentiated cell types (eg for long term memory). Sub-cellularly localised mRNAs must interact with specific RNA-binding proteins to ensure their normal localisation/ anchoring. Typically they are also under translational controls, to ensure they are only translated when at the target location. These phenomena have been studied in several systems, most notably in developmental patterning, cell migration and neurons. However many sub-cellularly localised mRNAs have been discovered through high-throughput studies, and the mechanisms underlying their localisation and translation have not been investigated. We have discovered a set of localised mRNAs, which are found specifically at the growing ends of Drosophila spermatids, in patterns resembling comets or cups. We also discovered a set of known RNA-binding proteins that also localise to this region. In this project you will investigate potential roles of the RNA-binding proteins in localising these specific mRNAs. RNA localisation and translation will be investigated in RNA-binding protein mutants. Protein-RNA interactions will be assayed in vitro with both purified components and extracts (to allow ternary complex formation). You will also determine whether, and how, mutations in the localised mRNAs and RNA-binding proteins affect the intricate structure of developing spermatid tail tips. This will provide the basis for a further analysis of this novel and virtually uncharacterised set of localised mRNAs. For example, systems biology and mathematical modelling approaches can be applied once the basic parameters of localisations, using super-resolution methods, and interactions at the biochemical and functional levels have been determined. Objectives - To describe and compare the comet and cup mRNAs', and RNA-binding proteins', localisations at the growing ends of spermatids. - To determine whether the known RNA-binding proteins are important for localisation of any comet and cup mRNAs. - To identify and characterise direct (or indirect) protein-RNA binding interactions between the localised mRNAs and the RNA-binding proteins. - To investigate whether mutations in comet and cup genes, and the RNA-binding proteins, cause defects in the cellular structure at the growing ends of elongating spermatids. - To uncover the relationship between "comet" and "cup" transcript localisation patterns. - To determine whether the known RNA-binding proteins regulate comet and cup mRNA translation.

信使核糖核酸定位于特定的亚细胞区域是极化细胞中的一种普遍现象，在发育过程(如构图)和分化细胞类型(如长时记忆)中都是至关重要的。亚细胞定位的mRNAs必须与特定的RNA结合蛋白相互作用，以确保它们的正常定位/锚定。通常情况下，它们也受平移控制，以确保它们仅在目标位置时才被翻译。这些现象已经在几个系统中进行了研究，最显著的是在发育模式、细胞迁移和神经元中。然而，通过高通量研究发现了许多亚细胞定位的mRNAs，而其定位和翻译的机制尚未被研究。我们已经发现了一组定位的mRNAs，它们专门在果蝇精子细胞的生长末端发现，其模式类似于彗星或杯子。我们还发现了一组已知的RNA结合蛋白，它们也定位于这个区域。在本项目中，您将研究RNA结合蛋白在定位这些特定mRNAs中的潜在作用。RNA定位和翻译将在RNA结合蛋白突变体中进行研究。蛋白质-RNA相互作用将在体外用纯化的成分和提取物进行分析(以形成三元复合体)。你还将确定局部mRNAs和RNA结合蛋白的突变是否以及如何影响发育中的精子细胞尾端的复杂结构。这将为进一步分析这一新颖的、几乎没有特征的局部化mRNA集提供基础。例如，一旦利用超分辨方法确定了定位的基本参数，并确定了生化和功能层面的相互作用，就可以应用系统生物学和数学建模方法。目的-描述和比较精子细胞生长末端的彗星和杯状mRNAs以及RNA结合蛋白的定位。-确定已知的RNA结合蛋白对任何彗星和杯状mRNAs的定位是否重要。-鉴定和表征定位的mRNAs和RNA结合蛋白之间的直接(或间接)蛋白质-RNA结合作用。-研究彗星和CUP基因以及RNA结合蛋白的突变是否会导致细长精子细胞生长末端的细胞结构缺陷。-揭示“彗星”和“杯”文字记录本地化模式之间的关系。-确定已知的RNA结合蛋白是否调节彗星和杯mRNA的翻译。