Lalaouna David, Prévost Karine, Park Seongjin, Chénard Thierry, Bouchard Marie-Pier, Caron Marie-Pier, Vanderpool Carin K, Fei Jingyi, Massé Eric
CRCHUS, RNA Group, Department of Biochemistry and Functional Genomics, Faculty of Medicine and Health Sciences, Université de Sherbrooke, 3201 Jean Mignault Street, Sherbrooke, QC J1E 4K8, Canada.
RNA architecture and reactivity Unit, Université de Strasbourg, CNRS, ARN UPR 9002, F-67000 Strasbourg, France.
Noncoding RNA. 2021 Sep 28;7(4):64. doi: 10.3390/ncrna7040064.
Many RNA-RNA interactions depend on molecular chaperones to form and remain stable in living cells. A prime example is the RNA chaperone Hfq, which is a critical effector involved in regulatory interactions between small RNAs (sRNAs) and cognate target mRNAs in Enterobacteriaceae. While there is a great deal of in vitro biochemical evidence supporting the model that Hfq enhances rates or affinities of sRNA:mRNA interactions, there is little corroborating in vivo evidence. Here we used in vivo tools including reporter genes, co-purification assays, and super-resolution microscopy to analyze the role of Hfq in RyhB-mediated regulation, and we found that Hfq is often unnecessary for efficient RyhB:mRNA complex formation in vivo. Remarkably, our data suggest that a primary function of Hfq is to promote RyhB-induced cleavage of mRNA targets by RNase E. Moreover, our work indicates that Hfq plays a more limited role in dictating regulatory outcomes following sRNAs RybB and DsrA complex formation with specific target mRNAs. Our investigation helps evaluate the roles played by Hfq in some RNA-mediated regulation.
许多RNA-RNA相互作用依赖分子伴侣在活细胞中形成并保持稳定。一个典型例子是RNA伴侣Hfq,它是参与肠杆菌科中小RNA(sRNA)与同源靶标mRNA之间调控相互作用的关键效应因子。虽然有大量体外生化证据支持Hfq提高sRNA:mRNA相互作用速率或亲和力的模型,但体内确证证据很少。在这里,我们使用包括报告基因、共纯化分析和超分辨率显微镜在内的体内工具来分析Hfq在RyhB介导的调控中的作用,并且我们发现Hfq在体内高效形成RyhB:mRNA复合物时通常并非必需。值得注意的是,我们的数据表明Hfq的主要功能是促进RyhB诱导的RNase E对mRNA靶标的切割。此外,我们的工作表明,在sRNA RybB和DsrA与特定靶标mRNA形成复合物后,Hfq在决定调控结果方面所起的作用更为有限。我们的研究有助于评估Hfq在某些RNA介导的调控中所起的作用。