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）： Messenger 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学和功能性细胞极化。极化细胞在成纤维细胞中的运动能力和树突中的突触可塑性，出现酵母的不对称分裂以及Xenopus和果蝇中的胚胎轴形成都需要从局部mRNA合成的蛋白质。迄今为止，对MRNA定位的研究集中在某些细胞类型中相对少量的mRNA上。然而，最近的一项研究发现了果蝇胚胎上皮中具有不同亚细胞分布的1500多种转录本，表明mRNA定位比以前想象的要普遍得多。然而，尚不清楚在专门的，形态复杂的细胞中进行定位的mRNA的完整曲目。 该竞争性修订应用程序是针对Not-OD-09-058的响应：“ NIH宣布了竞争性修订应用程序的恢复ACT资金的可用性”，建议检验以下假设：许多mRNA通过新型的基因组宽屏幕固定在多样化的专用细胞类型中，该基因组宽屏幕具有非含量亚纤维亚细胞分布的转录本。这个主要的新目标扩大了当前资助的赠款（R01 GM067758）的范围，该赠款的重点是使用果蝇纳米mRNA作为模型阐明mRNA定位的机制。屏幕结合了实验室中开发的一种用于MRNA在体内荧光标记的方法，其基因捕获和组织特异性基因表达的方法已建立。初始屏幕将集中在神经元细胞上，但将来可以扩展到其他细胞类型。具有组织特异性分布的mRNA的鉴定不仅可以洞悉细胞形态发生机制，而且还将为生物信息学分析提供数据库，以识别控制局部mRNA分布的顺式作用序列和/或结构基序。 公共卫生相关性：Messenger RNA定位会在发育过程中产生胚胎轴形成和不对称细胞分裂所需的局部蛋白质分布，以及在成纤维细胞中的偏振细胞功能以及树突中的突触可塑性。在分化细胞中进行定位的使者RNA的调节改变与各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。拟议的研究将阐明机制，通过这种机制，通过这种机制，将发育，生长和分化所需的基本细胞形态发生过程进行局部化，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。