Translational Regulation of Patterning in Drosophila

果蝇图案化的翻译调控

基本信息

批准号：
8643240
负责人：
ELIZABETH R GAVIS
金额：
$ 30.98万
依托单位：
PRINCETON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-05-01 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8643240
关键词：
3&apos Untranslated Regions Abdomen Affect Afferent Neurons Affinity Chromatography Animals Anterior Apoptotic Binding Biochemical Biochemical Genetics Cell Cycle Regulation Cell physiology Cells Chest Complement Complex Cytoplasm Data Dendrites Development Developmental Process Differentiation and Growth Disease Dissection Drosophila genus Elements Embryo Embryonic Development Event Gene Expression Gene Expression Regulation Gene Silencing Genes Genetic Translation Germ Grant Growth and Development function Head In Vitro Indium Investigation Light Malignant Neoplasms Mediating Messenger RNA Methods Modeling Molecular Morphogenesis Mutation Neurologic Dysfunctions Neurons Oocytes Oogenesis Ovary Pattern Phenotype Play Post-Transcriptional Regulation Process Protein Biosynthesis Proteins RNA RNA Interference RNA-Binding Proteins RNA-Protein Interaction Recovery Regulation Regulatory Element Repression Repressor Proteins Research Ribosomes Role Staging System Testing Transcript Transgenic Organisms Translation Initiation Translational Regulation Translational Repression Translations Work base egg flexibility genetic regulatory protein in vivo insight neuronal patterning polarized cell prevent protein complex protein structure function public health relevance research study stem

项目摘要

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 often depends on sequences found within their 3' untranslated regions (3'UTRs). Identification of proteins that interact with these 3'UTR regulatory elements has begun to shed light on the diversity of mechanisms that control translation. Recent studies indicate that multiple modes of translational regulation can be imposed on a transcript by different 3'UTR-binding factors, but how RNA-protein interactions affect such layers of control is poorly understood. The proposed research aims to elucidate translational control mechanisms underlying developmental processes, using the Drosophila nanos (nos) gene as a model. nos is ideal these studies, as nos encodes a translational repressor whose own synthesis must be highly regulated for proper embryonic development. During late oogenesis and early embryogenesis, nos translation must be repressed within the bulk cytoplasm for patterning of the anterior-posterior body axis. This repression is mediated by a translational control element (TCE) in the nos 3'UTR comprising two stem-loops that function in oogenesis and embryogenesis, respectively, through their interaction with the repressor proteins Glorund (Glo) and Smaug (Smg). In Aim 1, biochemical experiments that take advantage of a robust in vitro translation system based on ovary extract and a new method for ribosome footprinting are proposed to investigate the molecular mechanisms by which the TCE-Glo interaction inhibits translation by targeting both initiation and post-initiation events. This effort is supported by the identification and characterization of natie nos RNA-protein complexes and Glo interacting proteins proposed in Aim 2. In the early embryo, Nos protein forms a translational repressor complex with its partner Pumilio (Pum) to silence genes that inhibit abdominal and germline development. Identification of roles for Nos, Pum, and numerous additional RNA-binding proteins in neuronal morphogenesis during the previous grant period motivate biochemical and genetic studies proposed in Aim 3 to investigate the neuronal functions of these regulators and how post-transcriptional mechanisms modulate cellular processes that underlie morphogenesis and function of highly polarized cells. Mutations in translational regulatory proteins have been associated with a variety of cancers and diseases with neurological dysfunction. 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 disease.

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。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通常取决于在其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鉴定已经开始阐明控制翻译的机制的多样性。最近的研究表明，可以通过不同的3'UTR结合因子对转录本施加多种翻译调节模式，但是RNA-蛋白质相互作用如何影响这种控制层的了解很少。拟议的研究旨在以果蝇纳米（NOS）基因作为模型来阐明发展过程的转化控制机制。 NOS是理想的这些研究，因为NOS编码了一个翻译阻遏物，其自身的合成必须高度调节以进行适当的胚胎发育。在晚期的卵子发生和早期胚胎发生过程中，必须在整体细胞质中抑制NOS翻译，以使体内轴形成图案。这种抑制是由NOS 3'UTR中的翻译控制元件（TCE）介导的，该元素包括两个在卵子发生和胚胎发生中起作用的茎环，它们通过与抑制剂蛋白Glorund（Glo）和Smaug（Smg）（SMG）的相互作用。在AIM 1中，提出了利用基于卵巢提取物的强大体外翻译系统的生化实验，并提出了一种新的核糖体足迹方法来研究TCE-GLO相互作用通过同时启动和放电后靶向TCE-GLO相互作用来抑制翻译的分子机制。 Natie NOS NOS RNA-蛋白复合物和ALIC 2中提出的GLO相互作用蛋白的鉴定和表征支持了这一努力。在早期的胚胎中，NOS蛋白与其伴侣Pumilio（PUM）形成了一种抑制腹部和生殖线发育的转化抑制剂复合物。在上一个赠款期间，在神经元形态发生中的NOS，PUM和许多其他RNA结合蛋白的作用鉴定，在AIM 3中提出的生化和遗传学研究旨在研究这些调节剂的神经元功能以及后转录机制的神经元功能调节细胞如何调节高度层状细胞和高度偏光化的细胞的细胞过程。翻译调节蛋白的突变与神经功能障碍的多种癌症和疾病有关。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疾病的基本见解。