Genome-wide regulatory network governing neuronal mRNA translation.

描述（由申请人提供）：该提案旨在开发一种全基因组体内方法，以定义神经系统中控制mRNA翻译的一套调控相互作用。大脑的一个基本属性是感知体验驱动神经元之间突触连接的数量和强度的变化。这些突触修饰被认为是记忆和认知的神经相关物，需要在单个突触位点合成特定蛋白质以响应神经活动。控制突触蛋白的这种局部合成的机制知之甚少，但它的中断可能会产生严重的后果。该提案描述了一种方法，该方法将为每个基因提供神经翻译的个体遗传报告，并将确定一套调控机制。该方法涉及几个步骤。第一种方法是为每个基因创建体内翻译报告基因。第二种是高通量策略，以确定其翻译由特定调节器控制的基因集。作为原理的证明，我们将定义5个关键翻译调节因子的靶点，包括脆性X蛋白。脆性X染色体是最常见的遗传性智力低下。该提案还将产生一种资源，使研究界能够确定任何翻译调节剂的目标。我们的方法利用了果蝇模型系统的特殊遗传可操作性。基因调控机制和功能的高度保守性确保了我们所学到的大部分东西可以转移到人类身上。

DESCRIPTION (provided by applicant): This proposal seeks to develop a genome-wide in vivo approach to define the suite of regulatory interactions controlling mRNA translation in the nervous system. A fundamental property of the brain is that perceptual experiences drive modifications in number and strength of synaptic connections among neurons. These synapse modifications, which are thought to be neural correlates of memory and cognition, require synthesis of specific proteins at individual synaptic sites in response to neural activity. The mechanisms governing this local synthesis of synaptic proteins are poorly understood, but its disruption can have severe consequences. This proposal describes a method that will provide individual genetic reporters of neural translation for each gene, and will identify the suite of regulatory mechanisms. The approach involves several steps. First is a method to create in vivo reporters of translation for each gene. Second is a high-throughput strategy to identify the set of genes whose translation is governed by a particular regulator. As a proof of principle, we will define the targets of 5 key translational regulators, including Fragile-X protein. Fragile-X is the most common inherited form of mental retardation. This proposal also will generate a resource that will empower the research community to identify the set of targets for any translational regulator. Our method takes advantage of the exceptional genetic manipulability of the Drosophila model system. The high degree of conservation of gene regulatory mechanisms and function ensures that much of what we learn will be transferable to humans.