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模拟FGF-9 5'-UTRRNA的结构和DAP5结合部位,用于独立于帽子的翻译

Amanda Whittaker^1,2, Dixie J Goss^3,4

¹PhD Program in Biochemistry, The Graduate Center, CUNY, New York, New York 10016, USA.

RNA (New York, N.Y.)

|June 12, 2024

概括

顶部独立的翻译启动依赖于像DAP5这样的脚手架蛋白质与RNA结构结合. 这项研究揭示了三级RNA折叠,而不是序列,形成了DAP5结合点在开始编码子附近.

科学领域:

分子生物学分子生物学
在RNA生物学,RNA生物学.
生物化学生化学

背景情况:

基因表达的独立转化启动对于基因表达至关重要,通常由基因架构蛋白质,如真核细胞启动因子4G (eIF4G) 或其同类,死亡相关蛋白5 (DAP5) 介导.
在5'未翻译区域 (5' UTRs) 内的特定RNA结构促进了核糖体的招募以启动翻译,特别是病毒和细胞mRNA利用内部核糖体进入点 (IRES) 类似的机制.
虽然已知RNA结构对于病毒mRNA翻译很重要,但DAP5与细胞5'UTR相互作用的精确结合位和结构要求在很大程度上仍未定义.

研究的目的:

阐明DAP5与纤维细胞生长因子9 (FGF-9) mRNA的5' UTR结合的结构基础.
为了确定FGF-9 5' UTR中介于DAP5相互作用的特定核酸和结构特征.
为了调查RNA的三级结构,而不是保存序列或二级结构,是否构成DAP5.5的结合部位.

主要方法:

使用SHAPE-seq对FGF-9 5'-UTRRNA的二次结构建模 (通过原始扩展测序分析的选择性2'-基化).
生物化学分析包括DAP5足迹,脚印和UV交叉链接,以映射DAP5-RNA相互作用.
三级结构建模以可视化预测RNA结构内的DAP5结合核酸的空间排列.

主要成果:

SHAPE-seq分析揭示了186nt FGF-9 5'-UTR.的复杂的二次结构.
在DAP5足迹,脚印和交叉链接实验中,精确地确定了参与DAP5结合的核酸.
对FGF-9 5'-UTR三级结构的建模表明,DAP5与RNA的一面上聚集的核酸相互作用,靠近起始编码子.

结论:

在FGF-9 5' UTR上DAP5的结合部位是由其三级结构决定的,而不是特定的序列或二级结构图案.
DAP5似乎与靠近翻译起点的RNA区域相互作用,从而影响独立于cap的翻译.
这些发现表明,结构性相互作用,而不是特定序列的相互作用,是独立于上限的转化增强剂的关键,提供潜在的治疗点.

关键词:

DAP5 DAP5 的意思是DAP5 的意思.在DAP5结合部位.在FGF-9RNA中.生物物理学的生物物理学.顶值独立的翻译

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