Mechanism of RNA Localization in Drosophila Development

果蝇发育中RNA定位的机制

• 批准号: 7807579
• 负责人: ELIZABETH R GAVIS
• 金额: $ 45.78万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7807579
• 关键词: Afferent Neurons; Bioinformatics; Cell Polarity; Cell division; Cell physiology; Cells; Cellular Morphology; Cis-Acting Sequence; Complex; Databases; Defect; Dendrites; Destinations; Development; Differentiation and Growth; Disease; Drosophila genus; Embryo; Epithelium; Fibroblasts; Funding; Future; Genes; Grant; Growth and Development function; Investigation; Label; Lead; Light; Malignant Neoplasms; Messenger RNA; Methods; Modeling; Neurologic Dysfunctions; Neurons; Play; Process; Proteins; RNA; Recovery; Regulation; Role; Saccharomycetales; Signal Transduction; Synaptic plasticity; Testing; Tissue-Specific Gene Expression; Tissues; Transcript; United States National Institutes of Health; Xenopus; cell fate specification; cell motility; cell type; follow-up; genome wide association study; genome-wide; in vivo; insight; migration; novel; polarized cell; protein distribution; public health relevance; response

DESCRIPTION (provided by applicant): Messenger RNA (mRNA) localization has emerged as an important mechanism for generating asymmetric protein distributions that promote morphological and functional cell polarization during development. Polarized cellular functions like motility in fibroblasts and synaptic plasticity in dendrites, asymmetric division of budding yeast, and embryonic axis formation in Xenopus and Drosophila all require proteins that are synthesized from localized mRNAs. To date, investigations of mRNA localization have focused on a relatively small number of mRNAs in select cell types. However, a recent study uncovered over 1500 transcripts with distinct subcellular distributions in the Drosophila embryonic epithelium, indicating that mRNA localization is far more prevalent than previously thought. The full repertoire of mRNAs that undergo localization in specialized, morphologically complex cells is not known, however. This competitive revision application, in response to NOT-OD-09-058: "NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications", proposes to test the hypothesis that numerous mRNAs are localized in a diversity of specialized cell types through a novel genome wide-screen for transcripts with asymmetric subcellular distributions. This major new objective expands the scope of the currently funded grant (R01 GM067758), which focuses on elucidating mechanisms of mRNA localization using Drosophila nanos mRNA as a model. The screen combines a method developed in the lab for in vivo fluorescent labeling of mRNAs with well established methods for gene trapping and tissue specific gene expression. The initial screen will focus on neuronal cells but, in the future, can be expanded to other cell types. Identification of mRNAs with tissue-specific polarized distributions will not only provide insight into cellular morphogenetic mechanisms but will also provide a data-base for bioinformatic analysis to discern cis-acting sequences and/or structural motifs that govern localized mRNA distributions. PUBLIC HEALTH RELEVANCE: Messenger RNA localization produces localized protein distributions required for embryonic axis formation and asymmetric cell division during development and for polarized cellular functions like motility in fibroblasts and synaptic plasticity in dendrites. Altered regulation of messenger RNAs that undergo localization in differentiated cells have been associated with a variety of cancers as well as neuronal defects and may reflect roles for messenger RNA localization in cell fate specification, cell polarity, and migration. The proposed studies will shed light on mechanisms by which localized mRNAs control of basic cellular morphogenetic processes needed for development, growth, and differentiation and how the disruption of these processes may lead to diseases like cancer or neurological dysfunction.

描述（由申请人提供）： 信使 RNA (mRNA) 定位已成为产生不对称蛋白质分布的重要机制，从而促进发育过程中形态和功能细胞的极化。极化细胞功能，如成纤维细胞的运动性和树突的突触可塑性、芽殖酵母的不对称分裂以及非洲爪蟾和果蝇的胚胎轴形成，都需要由局部 mRNA 合成的蛋白质。迄今为止，mRNA 定位的研究主要集中在选定细胞类型中相对少量的 mRNA。然而，最近的一项研究在果蝇胚胎上皮中发现了超过 1500 个具有不同亚细胞分布的转录本，表明 mRNA 定位比之前想象的要普遍得多。然而，在专门的、形态复杂的细胞中进行定位的 mRNA 的全部功能尚不清楚。 此竞争性修订申请是为了回应 NOT-OD-09-058：“NIH 宣布恢复法案基金可用于竞争性修订申请”，提议通过新型基因组宽屏筛选具有不对称亚细胞分布的转录本来测试大量 mRNA 定位于多种特殊细胞类型的假设。这一重大新目标扩大了目前资助的资助范围 (R01 GM067758)，重点是使用果蝇 nanos mRNA 作为模型来阐明 mRNA 定位的机制。该屏幕将实验室开发的 mRNA 体内荧光标记方法与成熟的基因捕获和组织特异性基因表达方法相结合。最初的筛选将集中于神经元细胞，但将来可以扩展到其他细胞类型。具有组织特异性极化分布的 mRNA 的鉴定不仅可以深入了解细胞形态发生机制，而且还可以为生物信息学分析提供数据库，以辨别控制局部 mRNA 分布的顺式作用序列和/或结构基序。 公共健康相关性：信使 RNA 定位产生胚胎轴形成和发育过程中不对称细胞分裂以及极化细胞功能（如成纤维细胞运动和树突突触可塑性）所需的局部蛋白质分布。在分化细胞中定位的信使 RNA 的改变调节与多种癌症以及神经元缺陷有关，并且可能反映了信使 RNA 定位在细胞命运规范、细胞极性和迁移中的作用。拟议的研究将揭示局部 mRNA 控制发育、生长和分化所需的基本细胞形态发生过程的机制，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。