Translational Regulation of Patterning in Drosophila

果蝇图案化的翻译调控

• 批准号: 7650608
• 负责人: ELIZABETH R GAVIS
• 金额: $ 32.2万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7650608
• 关键词: 3' Untranslated Regions; Abdomen; Address; Affect; Afferent Neurons; Binding; Biochemical; Biochemical Genetics; Biological Assay; Cell Cycle; Cell Cycle Regulation; Cell physiology; Cells; Complement; Complex; Cues; Defect; Dendrites; Development; Developmental Process; Differentiation and Growth; Disease; Dissection; Drosophila Proteins; Drosophila genus; Elements; Embryo; Embryonic Development; Event; Fibrinogen; Funding; Gene Expression Regulation; Genetic Translation; Genome; Growth Factor Oncogenes; Growth and Development function; Image; Immune response; Indium; Investigation; Lead; Life; Light; Malignant Neoplasms; Mediating; Mediator of activation protein; Messenger RNA; Modeling; Molecular; Morphogenesis; Neurologic Dysfunctions; Neurons; Oocytes; Pattern; Play; Post-Transcriptional Regulation; Process; Protein Biosynthesis; Proteins; RNA; RNA Interference; RNA-Binding Proteins; RNA-Protein Interaction; Regulation; Regulatory Element; Repression; Research; Role; Staging; Structure; Transgenic Organisms; Translational Regulation; Translational Repression; Translations; Untranslated Regions; Work; base; cytokine; genetic regulatory protein; genome-wide; in vivo; insight; mutant; neuron development; neuronal patterning; protein complex; public health relevance; research study; response; stem; translation factor; tumorigenesis

DESCRIPTION (provided by applicant): Control of mRNA translation plays a key role in temporal and spatial regulation of gene expression during development. Translational regulation of mRNAs involved in a variety of developmental processes, including oocyte polarization, cell cycle regulation, embryonic patterning and neuronal morphogenesis, has been shown to depend on sequences found within their 3' untranslated regions (3'UTRs). Ongoing identification of proteins that interact with these 3'UTR regulatory elements reveals that mRNAs often employ multiple mechanisms to regulate translation, but how these mechanisms are coordinated is poorly understood. The abundance of RNA-binding proteins in metazoan genomes suggests that these and other mechanisms of post-transcriptional control are widespread. The proposed research aims to elucidate translational control mechanisms underlying developmental processes, using the Drosophila translational repressor Nanos (Nos) as a model. Nos, together with its partner Pumilio (Pum), directs abdominal and germline development during early embryogenesis and functions later during larval development for neuronal morphogenesis. Nos provides a unique opportunity for investigating post-transcriptional regulation of developmental events because translation of nos mRNA is itself highly regulated. Translational repression of nos mRNA is mediated by a bipartite translational control element (TCE) in the nos 3'UTR, comprising two stem-loops that function at different developmental stages via distinct binding factors. Aims 1 and 2 focus on TCE-mediated repression of nos mRNA to investigate how such RNA-protein interactions effect multiple layers of translational control: Aim 1 proposes transgenic assays and biochemical characterization of translationally repressed nos mRNA-protein complexes to determine how the TCE and its regulatory factors impact the translational apparatus while Aim 2 dissects the specific contribution of one factor, Glorund. Recent studies highlighting the relevance of translational control to neuronal morphogenesis motivate new directions in Aims 3 and 4 to investigate the roles of Nos, Pum, and other RNA-binding proteins: In Aim 3, live imaging and RNA target isolation approaches will be used to elucidate the role Nos/Pum- mediated regulation in dendrite morphogenesis; Aim 4 proposes an RNAi screen to uncover the underlying contribution of post-transcriptional gene regulation to neuronal development. These studies will lead to an understanding of how 3'UTR sequences and RNA-binding proteins regulate translation during development. More generally, they will shed light on mechanisms by which RNA- protein interactions provide the highly selective control of basic cellular processes needed for development, growth, and differentiation and how the disruption of these process may lead to diseases like cancer or neurological dysfunction. PUBLIC HEALTH RELEVANCE: Control of mRNA translation enables cells to alter protein synthesis rapidly and reversibly in response to external cues. Cell cycle mediators, oncogenes, growth factors, and cytokines have been shown to be translationally regulated and defects that alter levels of proteins that control translation are associated with tumorigenesis and disruption of the immune response. By investigating translational regulatory factors and the mechanisms by which they function, the proposed work will provide fundamental insight into how these factors control development, growth and differentiation, and how the disruption of translational control can result in diseases such as cancer.

描述（由申请人提供）：对mRNA翻译的控制在发育过程中基因表达的时间和空间调节中起关键作用。已证明，参与各种发育过程的mRNA的翻译调节，包括卵母细胞极化，细胞周期调节，胚胎模式和神经元形态发生，已证明取决于在其3'未翻译区域中发现的序列（3'UTRS）。与这些3'UTR调节元件相互作用的蛋白质的持续鉴定表明，mRNA通常采用多种机制来调节翻译，但是如何对这些机制进行了协调的了解很少了解。后生基因组中的RNA结合蛋白的丰度表明，这些转录后控制的这些机制和其他机制广泛。拟议的研究旨在将果蝇转化阻遏物纳米（NOS）作为模型阐明发展过程的转化控制机制。 NOS及其伴侣Pumilio（PUM）指导幼虫形态发生的幼虫发育期间的早期胚胎发生和功能，指导腹部和种系发育。 NOS为调查发育事件的转录后调节提供了独特的机会，因为NOS mRNA的翻译本身是高度调节的。 NOS mRNA的翻译抑制是由NOS 3'UTR中的两部分翻译控制元件（TCE）介导的，其中包括两个通过不同的结合因子在不同发育阶段发挥作用的茎环。目的1和2关注TCE介导的NOS mRNA的抑制，以研究这种RNA-蛋白质相互作用如何影响转化控制的多个层：目标1提出了转基因测定和翻译抑制的NOS mRNA蛋白配合物的生化表征，以确定TCE及其调节因素如何影响转化式的转化设备，而目标是贡献的贡献。最近的研究强调了转化控制与神经形态发生的相关性，激发了目标3和4中的新方向，以研究NOS，PUM和其他RNA结合蛋白的作用：在AIM 3中，实时成像和RNA靶隔离方法将用于阐明NOS/PUM介导的NOS/PUM介导的调节法规在树突中的作用； AIM 4提出了一个RNAi筛选，以发现转录后基因调节对神经元发育的基本贡献。这些研究将导致对3'UTR序列和RNA结合蛋白如何调节发育过程中的翻译。更普遍地，他们将阐明RNA蛋白相互作用提供对发育，生长和分化所需的基本细胞过程的高度选择性控制的机制，以及这些过程的破坏如何导致癌症或神经功能障碍等疾病。公共卫生相关性：mRNA翻译的控制使细胞能够响应外部提示而快速和可逆地改变蛋白质合成。细胞循环介质，癌基因，生长因子和细胞因子已被证明是受翻译调节的，并且缺陷改变了控制翻译的蛋白质水平与肿瘤发生和免疫反应的破坏有关。通过研究翻译调节因素及其功能的机制，拟议的工作将提供有关这些因素如何控制发展，生长和分化以及转化控制的破坏如何导致癌症等疾病的基本见解。