Dissecting the mechanisms of translational regulation in Drosophila germ stem cells

剖析果蝇生殖干细胞的翻译调控机制

• 批准号: 1775122
• 金额: --
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1775122
• 关键词: Dissecting; mechanisms; translational; regulation; Drosophila

BackgroundSTAR (signal transduction and activation of RNA) proteins are KH-domaincontaining, RNA-binding proteins whose activity is regulated in response to signals and regulate RNA processes. One family member is Drosophila HOW, which is expressed in the brain, testes and muscles. Mutants exhibit slow heart rates, heart defects and male sterility. Orthologs have been shown to regulate mRNA translation eg. p53 mRNA in C.elegans. HOW is required for germ stem cell maintanence in Drosophila testes and binds to RNAs that regulate differentiation in testes.Objectives1)Identify mRNAs translationally regulated by HOW in testes2)Characterise molecular mechanism of translational regulation by HOW3)Understand role of HOW regulated translation events in germ cellsNoveltyTransciptome-wide studies of translation have only been possible in the last few years, with most determining regions of translation and translational efficiencies. Combining ribosome profiling with structural analysis and Drosophila genetics is an innovative strategy, to dissect the molecular mechanisms of translational control by a specific protein.TimelinessThe advent of ribosome profiling recently allowed global assessments of translational regulation. Improvements to the method, including ours (Aspden eLife 2014) now allow translational regulation by specific proteins to be measured. The dogma has been that the majority of gene expression regulation occurs at transcription but recent work has revealed mRNA translation is also a key point of regulation, eg binding of proteins to mRNAs alters the ability of ribosomes to initiate translation. Structural studies of STAR-proteins are currently at the point (including Edwards group), where mechanistic insight into how binding affects RNA activity can be achieved.Experimental Approach-Next Generation Sequencing to study translational control (ribosome profiling)-Biochemical structural methods to understand how RNA-binding occurs and how this affects the ribosome (X-ray crystallography)-Drosophila genetic analysis to assess how molecular interactions affect germ stem cell biology and fly fertility.

研究背景STAR（signal transduction and activation of RNA）蛋白是一类含有KH结构域的RNA结合蛋白，其活性受信号调节，并调节RNA的加工过程。其中一个家族成员是果蝇HOW，它在大脑、睾丸和肌肉中表达。突变体表现出心率缓慢，心脏缺陷和雄性不育。直系同源物已被证明可以调节mRNA翻译，例如。秀丽隐杆线虫中p53 mRNA。HOW是果蝇睾丸中生殖干细胞维持所必需的，并与调节睾丸分化的RNA结合。目的1）鉴定睾丸中how调控的mRNAs 2）通过how表征翻译调控的分子机制3）了解HOW调控的翻译事件在生殖细胞中的作用新颖性在过去几年中，全转录组的翻译研究才成为可能，具有大多数决定翻译和翻译效率的区域。将核糖体分析与结构分析和果蝇遗传学相结合是一种创新的策略，可以剖析特定蛋白质翻译调控的分子机制。对该方法的改进，包括我们的方法（Aspden eLife 2014），现在允许测量特定蛋白质的翻译调节。教条是大多数基因表达调控发生在转录时，但最近的工作表明mRNA翻译也是调控的关键点，例如蛋白质与mRNA的结合改变了核糖体启动翻译的能力。STAR蛋白的结构研究目前正处于（包括爱德华兹集团），实验方法--新一代测序技术研究翻译调控（核糖体分析）-了解RNA结合如何发生以及如何影响核糖体的生化结构方法（X射线晶体学）-果蝇遗传分析，以评估分子相互作用如何影响生殖干细胞生物学和苍蝇生育力。

项目成果

Optimization of Ribosome Footprinting Conditions for Ribo-Seq in Human and Drosophila melanogaster Tissue Culture Cells.

• DOI: 10.3389/fmolb.2021.791455
• 发表时间: 2021
• 期刊: Frontiers in molecular biosciences
• 影响因子: 5
• 作者: Douka K;Agapiou M;Birds I;Aspden JL
• 通讯作者: Aspden JL

Held out wings RNA binding activity in the cytoplasm during early spermatogenesis

在早期精子发生过程中，在细胞质中保持翅膀 RNA 结合活性

• DOI: 10.1101/2022.05.19.492257
• 发表时间: 2022
• 作者: Agapiou M

Ribosome heterogeneity in Drosophila melanogaster gonads through paralog-switching.

• DOI: 10.1093/nar/gkab606
• 发表时间: 2022-02-28
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Hopes T;Norris K;Agapiou M;McCarthy CGP;Lewis PA;O'Connell MJ;Fontana J;Aspden JL
• 通讯作者: Aspden JL