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通过DMS MaP-Seqq可视化的核糖体诱导的mRNA伪结相互作用

Preston A Kellenberger¹, Zhenwei Song¹, Xiao Heng¹

¹Department of Biochemistry, University of Missouri, Columbia, MO, USA.

bioRxiv : the preprint server for biology

|November 19, 2025

概括

核糖体通过特定的键稳定RNA伪结,但也使它们不依赖于其位置占用而变得不稳定. 这项研究强调了DMS MaP-Seqq.

科学领域:

分子生物学分子生物学
结构生物学结构生物学
病毒学病毒学

背景情况:

RNA动态对于生物功能至关重要,在翻译过程中,核糖体与各种RNA结构相互作用.
已知RNA伪结会阻碍核糖体读取框架的维护.
了解伪结节-核糖体相互作用是破译翻译忠实性的关键.

研究的目的:

为了研究由核糖体诱导的RNA伪结的构造变化.
描述甘黄叶病毒伪结 (ScYLV_PK) 的稳定性和展开机制.
建立二甲基硫酸盐探测与突变分析和测序 (DMS MaP-Seq) 作为研究核糖体-RNA相互作用的工具.

主要方法:

利用二甲基硫酸盐探测与突变分析和测序 (DMS MaP-Seq) 来分析RNA结构.
应用DMS MaP-Seq来检查糖黄叶病毒伪结 (ScYLV_PK) 在核糖体的存在下展开的情况.
研究了特定残留物和核糖体占用率 (A/P位) 在伪结结稳定性中的作用.

主要成果:

ScYLV_PK的螺旋连接由核糖体稳定,取决于在残留27的结.
核糖体破坏野生类型ScyLV_PK的稳定,独立于A/P位点占用.
DMS MaP-Seq精确地检测到由核糖体诱导的RNA结构变化.

结论:

DMS MaP-Seq是一种敏感的方法,用于检测核糖体介导的RNA结构动态.
特定的结构图案,包括在残留物27的键,决定了转化过程中伪结的稳定性.
核糖体与伪结的相互作用涉及复杂的稳定和不稳定机制.

关键词:

结构 RNA 结构 RNA 结构核糖体核糖体核糖体核糖体它们是mRNARNA.

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