G quartet RNA-FMRP interactions in Fragile X syndrome

脆性 X 综合征中 G 四重奏 RNA-FMRP 相互作用

• 批准号: 6954409
• 负责人: MIHAELA R MIHAILESCU
• 金额: $ 21.01万
• 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 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6954409
• 关键词: RNA; RNA binding protein; fragile X syndromes; intermolecular interaction; methylation; nerve /myelin protein; nucleic acid sequence; nucleic acid structure; protein structure; protein structure function; spectrometry; thermodynamics

DESCRIPTION (provided by applicant): This grant focuses on the investigation of the fragile X mental retardation protein (FMRP) interactions with G quartet forming RNA target(s). Fragile X syndrome is the most common form of inherited mental retardation, affecting ~ 1 in 4000 males and ~ 1 in 8000 females. The syndrome is caused by the loss of a normal cellular protein (FMRP), which is thought to act as a translational repressor of specific messenger RNA (mRNA). Although the in vivo RNA targets of the protein remain elusive, it has been reported that FMRP binds with high affinity to RNA sequences rich in guanine content, which fold into G quartet structures. Given the importance of the FMRP RNA binding activity for its function, one of the specific aims of this grant is to investigate the molecular basis of RNA recognition by FMRP. Is the G-quartet containing RNA recognized via a sequence specific, via a structure specific mechanism, or both? Is FMRP stabilizing or destabilizing these structures? FMRP binds the G quartet forming RNA using its arginine-glycine rich domain (RGG box), a domain found in many other RNA binding proteins. Another aim of the proposal is to determine the level of specificity with which the G quartet structures are recognized by this domain, by analyzing the interactions of 2 different RGG boxes derived from other RNA binding proteins with these RNA sequences. The third aim of the proposal is to determine if protein arginine methylation, a common posttranslational modification involving the methylation of arginine residues within the RGG box, plays a role in modulating the FMRP-RNA interactions. Answers to such questions will contribute to our ability to identify the in vivo targets of FMRP, in an effort to understand the link between the absence of the protein and the phenotype of the fragile X syndrome. To accomplish these goals, molecular biology and biochemistry methods will be used to produce the RNA and protein and biophysical techniques such as NMR spectroscopy, fluorescence spectroscopy, UV-Vis spectroscopy will be employed to study these biomolecules and their interactions.

描述（由申请人提供）：该赠款的重点是研究脆弱的X智力低下蛋白（FMRP）与G四重奏形成RNA靶标的相互作用。脆弱的X综合征是遗传性智力低下的最常见形式，影响了4000名男性中的约1个，而8000名女性中有约1个。该综合征是由正常细胞蛋白（FMRP）的丧失引起的，该蛋白质被认为是特定信使RNA（mRNA）的翻译阻遏物。尽管该蛋白质的体内RNA靶标仍然难以捉摸，但据报道，FMRP与富含鸟嘌呤含量的RNA序列具有高亲和力结合，它们折叠成G四重奏结构。鉴于FMRP RNA结合活性对其功能的重要性，该赠款的具体目的之一是研究FMRP识别RNA识别的分子基础。包含RNA的G-四分之一是通过序列特异性（通过结构特定机制）识别的？ FMRP是否稳定或破坏这些结构？ FMRP使用其精氨酸 - 甘氨酸富含结构域（RGG框）结合G四重奏形成RNA，这是一个在许多其他RNA结合蛋白中发现的结构域。该提案的另一个目的是通过分析来自其他RNA结合蛋白与这些RNA序列的2个不同的RGG框的相互作用来确定该域G四重奏结构的特异性水平。该提案的第三个目的是确定蛋白精氨酸甲基化是否是涉及RGG盒中精氨酸残基甲基化的常见翻译后修饰，在调节FMRP-RNA相互作用中起着作用。此类问题的答案将有助于我们识别FMRP体内靶标的能力，以了解缺乏蛋白质与脆弱X综合征的表型之间的联系。为了实现这些目标，将使用分子生物学和生物化学方法来生产RNA，蛋白质和生物物理技术，例如NMR光谱，荧光光谱法，UV-VIS光谱法研究这些生物分子及其相互作用。