Department of Biological Sciences, College of Biological Sciences and Biotechnology, Chungnam National Universitygrid.254230.2, Daejeon, Republic of Korea.
Infection Control Convergence Research Center, Chungnam National Universitygrid.254230.2 College of Medicine, Daejeon, Republic of Korea.
mBio. 2022 Jun 28;13(3):e0128722. doi: 10.1128/mbio.01287-22. Epub 2022 Jun 13.
In Escherichia coli, transcription is coupled with translation. The polar operon is transcribed ; however, about 10% of transcription terminates at the end of because of Rho-dependent termination (RDT). When translation is complete, translation should begin immediately. It is unclear whether RDT at the end of is due to decoupling of translation termination and transcription at the cistron junction. In this study, we show that RDT at the cistron junction is linked to the failure of translation initiation at the start of , rather than translation termination at the end of . We also show that transcription pauses 130 nucleotides downstream from the site of translation termination, and this pause is required for RDT. Transcription of operons is initiated at the promoter of the first gene in the operon, continues through cistron junctions, and terminates at the end of the operon, generating a full-length mRNA. Here, we show that Rho-dependent termination of transcription occurs stochastically at a cistron junction, generating a stable mRNA that is shorter than the full-length mRNA. We further show that stochastic failure in translation initiation of the next gene, rather than the failure of translation termination of the preceding gene, causes the Rho-dependent termination. Thus, stochastic failure in translation initiation at the cistron junction causes the promoter-proximal gene to be transcribed more than promoter-distal genes within the operon.
在大肠杆菌中,转录与翻译偶联。极性操纵子被转录;然而,由于 Rho 依赖性终止(RDT),约 10%的转录在 结束。当 翻译完成时, 翻译应该立即开始。尚不清楚 末端的 RDT 是否是由于密码子交界处的翻译终止和解偶联所致。在这项研究中,我们表明,密码子交界处的 RDT 与 起始处翻译起始的失败有关,而不是 末端的翻译终止。我们还表明,转录在 翻译终止位点的下游 130 个核苷酸处暂停,并且该暂停是 RDT 所必需的。操纵子的转录从操纵子中第一个基因的启动子开始,通过密码子交界处继续,并在操纵子的末端终止,生成全长 mRNA。在这里,我们表明转录在密码子交界处的 Rho 依赖性终止是随机发生的,生成比全长 mRNA 短的稳定 mRNA。我们进一步表明,下一个基因翻译起始的随机失败,而不是前一个基因翻译终止的失败,导致 Rho 依赖性终止。因此,密码子交界处翻译起始的随机失败导致启动子近端基因的转录比操纵子内启动子远端基因更多。