Investigation of Fragile X mental retardation protein interactions with the miRNA pathway.

脆性 X 智力迟钝蛋白与 miRNA 通路相互作用的研究。

• 批准号: 9303733
• 负责人: MIHAELA R MIHAILESCU
• 金额: $ 41.4万
• 依托单位: DUQUESNE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2021-09-20
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9303733
• 关键词: Affect; Affinity; Alternative Splicing; Arginine; Binding; Binding Sites; Biochemical; Biogenesis; Biological Process; Complex; Development; Dicer Enzyme; Event; Exons; FMR1; Female; Fragile X Syndrome; G-Quartets; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Glycine; Goals; Guide RNA; Impairment; Inherited; Investigation; KH Domain; Knowledge; Laboratories; Length; Mental Retardation; Messenger RNA; MicroRNAs; Molecular; Molecular Chaperones; Names; Nucleic Acids; Nucleotides; Pathway interactions; Phenotype; Phosphorylation; Play; Process; Production; Protein Biosynthesis; Protein Isoforms; Proteins; Psyche structure; RNA; RNA Interference; RNA Sequences; RNA-Binding Proteins; RNA-Induced Silencing Complex; Regulation; Research; Role; SUI1 gene; Site; Specificity; Structure; Synapses; Syndrome; Testing; Translations; Untranslated RNA; Work; male; novel; response

Project Summary Fragile X mental retardation syndrome is the most common form of inherited mental retardation, affecting ~ 1 in 3600 males and ~ 1 in 6000 females. The syndrome is caused by the loss of a normal cellular protein, named the fragile X mental retardation protein (FMRP). Despite extensive research in the past two decades, the relationship between the absence of FMRP and the phenotype of the fragile X syndrome is still not fully understood. FMRP is an RNA binding protein involved in the transport and translation regulation of specific messenger RNA (mRNA) targets. Biochemical studies have determined that FMRP uses its arginine-glycine- glycine (RGG) box to bind with high affinity to RNA sequences that form G quadruplex (GQ) structures. The mechanisms by which FMRP exerts its translation regulator function are not known, however it has been proposed that the protein works in conjunction with the microRNA (miRNA) pathway to regulate local protein synthesis in response to synaptic input. This proposal has the following specific aims: 1. Investigation of the FMRP role in miRNA maturation. Several miRNAs have been shown to require FMRP to exert their function, and FMRP has also been shown to bind pre-miRNAs. We hypothesize that FMRP regulates the maturation of specific miRNAs through its interactions with GQ structures formed by their precursor pre-miRNAs. To test this hypothesis we will characterize the GQ structures predicted to form in selected pre-miRNAs and analyze their interactions with FMRP. Subsequently, we will analyze the effect of FMRP upon the production of mature miRNAs by Dicer. 2. Investigation of FMRP: GQ mRNA: miRNA interactions in translation regulation. FMRP has been shown to directly regulate the translation of specific mRNAs through its interactions with the miRNA-guided RNA induced silencing complex (RISC). Following the guiding hypothesis that FMRP interacts with GQ structures in specific mRNAs to modulate their recognition by RISC, under this aim we will analyze FMRP: GQ mRNA: miRNA interactions in the context of other mRNAs that have predicted GQ structures: (i) within their miRNA binding sites and (ii) immediately adjacent to them. 3. Investigation of various FMRP isoforms interactions with the miRNA pathway. We hypothesize that the FMRP isoforms created by alternative splicing at exon 12 will have different interactions with miRNA precursors and/or protein components of the miRNA pathway, which could potentially result in functional differences with respect to translation regulation. Additionally, we hypothesize that FMRP activity-dependent primary and secondary phosphorylation events could further modulate these functional differences.

项目摘要 脆弱的X心理迟缓综合征是遗传性智力低下的最常见形式，影响〜1 在3600名男性中，有6000名女性中的1个。该综合征是由正常细胞蛋白的丧失引起的 命名为脆弱的X智障蛋白（FMRP）。尽管在过去的二十年中进行了广泛的研究，但 FMRP缺乏与脆弱X综合征的表型之间的关系仍然不完全 理解。 FMRP是一种参与特定运输和翻译调节的RNA结合蛋白 Messenger RNA（mRNA）靶标。生化研究确定FMRP使用其精氨酸 - 甘氨酸 - 甘氨酸（RGG）盒与高亲和力结合与形成G四链体（GQ）结构的RNA序列。这 FMRP发挥其翻译调节器函数的机制尚不清楚，但是它已经 提出该蛋白质与microRNA（miRNA）途径结合起来调节局部蛋白 响应突触输入的合成。该建议具有以下具体目标： 1。研究FMRP在miRNA成熟中的作用。已经证明了几个miRNA需要 FMRP发挥其功能，并且FMRP也已被证明结合了前MIRNA。我们假设这一点 FMRP通过其与其与其GQ结构相互作用的相互作用来调节特定miRNA的成熟 前体前体。为了检验该假设，我们将表征预测形成的GQ结构 选定的前MIRNA并分析其与FMRP的相互作用。随后，我们将分析 FMRP在DICER生产成熟的miRNA时。 2。FMRP的研究：GQ mRNA：翻译调节中的miRNA相互作用。 FMRP已显示 通过与miRNA引导的RNA的相互作用直接调节特定mRNA的翻译 诱导沉默复合物（RISC）。遵循指导假设，即FMRP与GQ结构相互作用 特定的mRNA调节RISC的认可，在此目的下，我们将分析FMRP：GQ mRNA： miRNA相互作用在其他已经预测GQ结构的mRNA的背景下：（i）在其mirna中 结合位点和（ii）紧邻它们。 3。研究各种FMRP同工型与miRNA途径的相互作用。我们假设 FMRP在外显子12上产生的替代剪接产生的同工型将与miRNA具有不同的相互作用 miRNA途径的前体和/或蛋白质成分，这可能会导致功能性 关于翻译调节的差异。此外，我们假设FMRP活动依赖性 原发性和继发磷酸化事件可以进一步调节这些功能差异。