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牛顿的摇篮般的全性机制解释了调节性RsmERNA结合的调节性RsmERNA结合

Esteban Finol¹, Fred F Damberger¹, Miroslav Krepl²

¹Institute for Biochemistry, Department of Biology, ETH Zurich, 8093, Zurich, Switzerland.

Research square

|May 5, 2025

概括

细菌非编码RNAs (ncRNAs) 通过隔离调节蛋白来调节基因翻译. 在Pseudomonas蛋白质中,RsmZ ncRNA使用负合作性来控制RsmE蛋白结合,促进mRNA释放.

科学领域:

细菌的基因调节细菌的基因调节
RNA与蛋白质的相互作用
分子机制的分子机制

背景情况:

Csr/Rsm系统通过非编码RNA (ncRNA) 和调节蛋白控制细菌中的mRNA翻译.
在Pseudomonas蛋白质中,RsmZ ncRNA充当"蛋白质海绵",隔离RsmE蛋白质二极体以调节翻译.
在RsmE结合过程中,RsmZ表现出异常的负合作性,在此过程中,初始结合会降低后续位点的亲和力.

研究的目的:

阐明RsmE与RsmZncRNA结合的负合作性背后的分子机制.
了解这种负合作性如何促进RsmE从mRNA释放.

主要方法:

异热定位热量计 (ITC) 用于测量结合热力学.
核磁共振 (NMR) 光谱检测结构变化.
分子动力学 (MD) 模拟以建模蛋白质-RNA相互作用和结构动力学.

主要成果:

最初的RsmE与RsmZ的结合事件增加了未被占用的结合位点的 conformational ,导致C端螺旋部分展开.
一种全osteric 机制将 RNA 结合在一个位点与另一个位点的形状变化结合在一起,解释了随后结合的 afinity 减少.
在RsmE二元体内的反平行β片介导了位点间的通信,涉及H键收缩和放松.

结论:

在RsmZ-RsmE相互作用中,负合作性是由全形状变化和驱动的.
这种机制可以有效地将RSME从mRNA转移到ncRNA,微调细菌基因表达.
这些发现揭示了RNA-蛋白相互作用中全性调节的新模式.

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