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RNA-RNA交互原子方法在体内提供了证据,证明Hfq边缘面在sRNA-mRNA配对中的关键作用

Xing Luo¹, Aixia Zhang², Caroline Esnault³

¹Laboratory of Molecular Biology, National Cancer Institute, Bethesda, MD 20892, USA.

bioRxiv : the preprint server for biology

|July 16, 2025

概括

细菌RNA陪伴者Hfq对于基因调节至关重要. 它的边缘面对于稳定RNA-RNA相互作用至关重要,其中的突变显著减少了基因配对.

科学领域:

分子生物学分子生物学
细菌遗传学细菌遗传学
在RNA生物学,RNA生物学.

背景情况:

细菌的小调节RNAs (sRNAs) 通过与目标信使RNAs (mRNAs) 的基因配对来控制基因表达.
配合RNA蛋白Hfq (宿主因子1) 对于许多sRNA-mRNA相互作用至关重要,具有不同的RNA结合面 (近位,边缘,远位).
Hfq促进sRNA和目标mRNA的同时结合,这对于基因调节至关重要.

研究的目的:

系统地研究Hfq的RNA结合面突变对活体RNA-RNA配对的功能影响.
阐明Hfq近接面,边缘面和远距离面在调解sRNA-mRNA相互作用中的特定作用.
为了比较不同RNA-RNA相互作用体方法的疗效,包括RIL-seq和iRIL-seq.

主要方法:

通过结合和测序 (RIL-seq) 方法利用RNA相互作用来系统地检查体内RNA-RNA相互作用.
引入了Hfq.的近端,边缘和远端RNA结合面的点突变.
使用细胞内RIL-seq (iRIL-seq) 来评估RNA-RNA相互作用,减少了体外处理步骤.

主要成果:

影响远端和近端Hfq结合面的突变者保留了显著的RNA-RNA相互作用 (S-chimeras).
边缘面突变 (R16A) 几乎完全丧失了S基梅拉,尽管保留了部分RNA结合和调节活性.
在R16A突变体上的iRIL-seq产生了比标准RIL-seq更多的S-基默,但比野生类型Hfq少,具有改变的流行RNA-RNA对.

结论:

Hfq的边缘面在活体中稳定RNA对中起着至关重要的作用.
不同的RNA-RNA互动原子方法捕捉了Hfq介导相互作用的不同方面.
了解Hfq的RNA结合面提供了对细菌基因调节机制的关键见解.

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