Genome-wide identification of mRNA localization motifs and factors

mRNA 定位基序和因子的全基因组鉴定

• 批准号: 8990859
• 负责人: Jefferson Matthew Taliaferro
• 金额: $ 5.8万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8990859
• 关键词: Address; Apical; Atrophic; Axon; Back; Binding; Binding Proteins; Cell Aging; Cell Nucleus; Cells; Chemotactic Factors; Cis-Acting Sequence; Clinical; Complex; Disease; Environment; Eukaryotic Cell; Family; Gene Expression; Genetic Transcription; Genome; Genomics; Goals; Growth Cones; Health; Imagery; Knowledge; Learning; Location; Long-Term Potentiation; MS2 coat protein; Mediating; Memory; Messenger RNA; Metabolism; Molecular; Molecular Genetics; Mus; Mutation; Nature; Neuroblastoma; Neurons; Prevalence; Process; Production; Protein Binding Domain; Protein Isoforms; Proteins; RNA; RNA Biochemistry; RNA Interference; RNA Sequences; Reaction Time; Regulation; Reporter; Side; Signal Transduction; Spinal Ganglia; Staging; Stimulus; Streptavidin; Structure; System; Therapeutic; Trans-Activators; Transcript; Translations; axon injury; base; beta Actin; cell age; cell type; experience; genome-wide analysis; neuronal growth; overexpression; response; transcriptome; transcriptome sequencing

DESCRIPTION (provided by applicant): Control of gene expression is regulated at many stages. One aspect of this regulation, the localization of specific mRNA transcripts to distinct cellular locations, has only recently been fully appreciated. Correct targeting of these messages can be crucial for many cellular and higher-order processes. For example, disruption of dendritic targeting of specific messages in mice leads to a decrease in long-term potentiation, implying that this process may be important for learning and memory. Recent studies have shown that up to two thousand mRNAs are localized to axons in neuronal cells. However, little is known about how messages are tagged for transport or the factors that mediate it. Although the mechanism of localization of a handful of messages has been dissected, how such mechanisms may operate on a transcriptome-wide scale is unknown. The studies outlined in this proposal aim to define the cis-acting sequences and trans-acting factors responsible for the localization of many messages. Specifically, I will address this question using a combination of RNA biochemistry, molecular genetics, and computational genomics. Using RNA-seq from axons and cell bodies, I will define the RNA sequences necessary for localization of specific messages. I will then refine these sequences using an MS2-GFP reporter system and search the localized transcriptome both computationally and experimentally for other instances of these motifs, whether they be contained in primary or secondary RNA structure. The factors that bind these motifs will then be identified biochemically and their effects on the localization of other messages will be identified using RNAi and localized RNA-seq. The results of these studies will allow the identification of families of messages that use similar motifs and factors for their localization, as well as the prediction of motif sequences in other messages. This has important clinical and therapeutic implications as mutations in many RNA-localizing proteins are associated with several neuronal diseases, including spinomuscular atrophy and ALS.

描述（由申请人提供）：基因表达的控制在许多阶段受到调节。这种调节的一个方面，特定的mRNA转录本定位到不同的细胞位置，最近才得到充分的理解。这些信息的正确定位对于许多细胞和高阶过程至关重要。例如，小鼠特定信息的树突靶向被破坏会导致长时程增强作用的减少，这意味着这个过程可能对学习和记忆很重要。最近的研究表明，多达2000个mRNA定位于神经元细胞的轴突。然而，很少有人知道的信息是如何标记的运输或调解它的因素。虽然定位的少数消息的机制已被解剖，如何这样的机制可能会在转录组范围内运作是未知的。本提案中概述的研究旨在定义负责许多信息定位的顺式作用序列和反式作用因子。具体来说，我将结合RNA生物化学、分子遗传学和计算基因组学来解决这个问题。使用来自轴突和细胞体的RNA-seq，我将定义定位特定信息所需的RNA序列。然后，我将使用MS 2-GFP报告系统来完善这些序列，并通过计算和实验来搜索这些基序的其他实例的本地化转录组，无论它们是否包含在初级或二级RNA结构中。然后将通过生物化学方法鉴定结合这些基序的因子，并鉴定它们对其他信息定位的影响 使用RNAi和localized RNA-seq。这些研究的结果将允许家庭的信息，使用类似的图案和因素，他们的本地化，以及在其他消息的基序序列的预测识别。这具有重要的临床和治疗意义，因为许多RNA定位蛋白的突变与几种神经元疾病相关，包括脊髓肌萎缩和ALS。