Genome-wide regulatory network governing neuronal mRNA translation.

描述(由申请人提供)：该提案寻求开发一种全基因组的活体方法来定义一套控制神经系统中信使核糖核酸翻译的调控相互作用。大脑的一个基本属性是知觉经验驱动神经元之间突触连接的数量和强度的改变。这些突触修饰被认为是记忆和认知的神经关联，需要在单个突触位置合成特定的蛋白质来响应神经活动。支配这种突触蛋白局部合成的机制尚不清楚，但它的破坏可能会产生严重的后果。这项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单独遗传报告，并将确定一套调控机制。该方法包括几个步骤。首先是一种为每个基因创建体内翻译记者的方法。其次是一种高通量策略，以识别其翻译受特定监管机构控制的一组基因。作为原则的证明，我们将定义包括脆性-X蛋白在内的5个关键翻译调控因子的靶标。脆性X是最常见的遗传性精神发育迟滞。这项提议还将产生一种资源，使研究界能够为任何翻译监管机构确定一套目标。我们的方法利用了果蝇模型系统特殊的遗传可操作性。基因调控机制和功能的高度保守确保了我们所学的许多东西将被转移到人类身上。 与公共健康相关：大脑的一个基本属性是，我们的经历会导致神经元之间连接的数量和强度发生变化。这些变化被认为是记忆和认知的基础，需要精确控制这些连接部位特定蛋白质的合成(1-5)。支配这种突触蛋白局部合成的机制尚不清楚，尽管我们确实知道这一过程的中断会产生严重的后果。这种缺陷被认为是脆性X综合征的原因。这项提议描述了一种方法，该方法可以为基因组中每个基因的蛋白质合成调控提供单独的报告，并可以确定控制每个基因的调控机制。

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. PUBLIC HEALTH RELEVANCE: A fundamental property of the brain is that our experiences cause modifications in number and strength of the connections among neurons. These changes, which are thought to underlie memory and cognition, require the precise control of the synthesis of specific proteins at the sites of these connections (1-5). The mechanisms governing this local synthesis of synaptic proteins are poorly understood, although we do know that disruption of this process has severe consequences. This defect is thought to be responsible for the Fragile-X syndrome. This proposal describes a method that can provide individual reporters of the regulation of protein synthesis for each gene in the genome, and can identify the regulatory mechanisms controlling each one.