Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, 2780-157 Oeiras, Portugal;
Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, 2780-157 Oeiras, Portugal.
Proc Natl Acad Sci U S A. 2019 Feb 19;116(8):3042-3051. doi: 10.1073/pnas.1811589116. Epub 2019 Feb 4.
Transcription termination is a critical step in the control of gene expression. One of the major termination mechanisms is mediated by Rho factor that dissociates the complex mRNA-DNA-RNA polymerase upon binding with RNA polymerase. Rho promotes termination at the end of operons, but it can also terminate transcription within leader regions, performing regulatory functions and avoiding pervasive transcription. Transcription of is autoregulated through a Rho-dependent attenuation in the leader region of the transcript. In this study, we have included an additional player in this pathway. By performing MS2-affinity purification coupled with RNA sequencing (MAPS), transcript was shown to directly interact with the small noncoding RNA SraL. Using bioinformatic in vivo and in vitro experimental analyses, SraL was shown to base pair with the 5'-UTR of mRNA upregulating its expression in several growth conditions. This base pairing was shown to prevent the action of Rho over its own message. Moreover, the results obtained indicate that both ProQ and Hfq are associated with this regulation. We propose a model that contemplates the action of SraL sRNA in the protection of mRNA from premature transcription termination by Rho. Note that since the interaction region between both RNAs corresponds to a very-well-conserved sequence, it is plausible to admit that this regulation also occurs in other enterobacteria.
转录终止是基因表达调控的关键步骤之一。主要的终止机制之一是由 Rho 因子介导的,当 Rho 因子与 RNA 聚合酶结合时,它会使 mRNA-DNA-RNA 聚合酶复合物解离。Rho 促进操纵子末端的终止,但它也可以在启动子区域内终止转录,从而发挥调节功能并避免普遍性转录。通过在转录本的启动子区域中依赖 Rho 的衰减,对 进行了自动调节。在本研究中,我们在这个途径中加入了一个额外的参与者。通过进行 MS2 亲和纯化结合 RNA 测序 (MAPS),发现 转录本与小非编码 RNA SraL 直接相互作用。通过生物信息学体内和体外实验分析,表明 SraL 与 mRNA 的 5'-UTR 碱基配对,在几种生长条件下上调其表达。这种碱基配对被证明可以防止 Rho 对其自身消息的作用。此外,所得结果表明 ProQ 和 Hfq 都与这种调节有关。我们提出了一个模型,考虑了 SraL sRNA 在保护 mRNA 免受 Rho 引发的过早转录终止方面的作用。请注意,由于两个 RNA 之间的相互作用区域对应于一个非常保守的序列,因此可以假定这种调节也发生在其他肠杆菌中。