结构知情的突变发生识别出一个对小鼠胰岛素受体5'UTR IRES功能至关重要的保存区域
bioRxiv : the preprint server for biology
|November 22, 2024
概括
细胞在压力时使用内部核糖体进入点 (IRES) 来进行翻译. 研究人员绘制了胰岛素受体5'非翻译区域 (Insr 5'UTR) 结构图,确定了一个关键区域的帽子独立翻译.
科学领域:
- 分子生物学分子生物学
- 遗传学 遗传学 是一个
- 生物化学 生化学
背景情况:
- 在压力期间,细胞抑制了卡普依赖的翻译.
- 内部核糖体进入点 (IRES) 能够在压力下翻译关键转录.
- 细胞IRES的RNA二次结构在很大程度上是未知的.
研究的目的:
- 在细胞中探测胰岛素受体5'非翻译区域 (Insr 5'UTR) IRES的二次结构.
- 确定对Insr IRES功能至关重要的结构元素.
- 设计一个最小的IRES元素,具有高上限的独立翻译活动.
主要方法:
- 通过测序 (DMS-MaPseq) 的二甲基硫酸盐突变概况被用于在体外和细胞内探测RNA二次结构.
- 采用了一种突变策略,以Insr 5'UTR的结构模型为指导.
- 病毒IRES (HCV,EMCV) 被用作对照.
主要成果:
- 细胞中的病毒IRES结构与已知的体外结构一致.
- 病毒IRESes的显著线性化发生在细胞中翻译起始码子附近.
- 确定了Insr 5'UTR的一个被保护的段落,距离起始编码子的距离,对于IRES活动至关重要.
结论:
- 使用DMS-MaPseq数据生成了Insr 5'UTR的结构模型.
- 设计了一个具有完全活动的最小的Insr IRES元素.
- 这项工作阐明了细胞IRES的结构和功能,有助于应激反应研究.
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