Nadiras Cédric, Schwartz Annie, Delaleau Mildred, Boudvillain Marc
Centre de Biophysique Moléculaire (UPR 4301), CNRS, rue Charles Sadron, Orléans, France.
Ecole doctorale Santé, Sciences Biologiques et Chimie du Vivant (ED 549), Université d'Orléans, Orléans, France.
Methods Mol Biol. 2018;1737:99-118. doi: 10.1007/978-1-4939-7634-8_7.
Besides their well-known posttranscriptional effects on mRNA translation and decay, sRNAs and associated RNA chaperones (e.g., Hfq, CsrA) sometimes regulate gene expression at the transcriptional level. In this case, the sRNA-dependent machinery modulates the activity of the transcription termination factor Rho, a ring-shaped RNA translocase/helicase that dissociates transcription elongation complexes at specific loci of the bacterial genome. Here, we describe biochemical assays to detect Rho-dependent termination signals in genomic regions of interest and to assess the effects of sRNAs and/or associated RNA chaperones on such signals.
除了对mRNA翻译和降解具有众所周知的转录后作用外,小RNA(sRNA)和相关的RNA伴侣(如Hfq、CsrA)有时也会在转录水平上调节基因表达。在这种情况下,依赖sRNA的机制会调节转录终止因子Rho的活性,Rho是一种环状RNA转位酶/解旋酶,可在细菌基因组的特定位点解离转录延伸复合物。在这里,我们描述了生化分析方法,用于检测感兴趣基因组区域中依赖Rho的终止信号,并评估sRNA和/或相关RNA伴侣对此类信号的影响。