Institute of Microbiology and Molecular Biology, University of Gießen, Gießen, Germany.
J Bacteriol. 2014 Apr;196(7):1435-47. doi: 10.1128/JB.01471-13. Epub 2014 Jan 31.
Quorum sensing of Sinorhizobium meliloti relies on N-acyl-homoserine lactones (AHLs) as autoinducers. AHL production increases at high population density, and this depends on the AHL synthase SinI and two transcriptional regulators, SinR and ExpR. Our study demonstrates that ectopic expression of the gene rne, coding for RNase E, an endoribonuclease that is probably essential for growth, prevents the accumulation of AHLs at detectable levels. The ectopic rne expression led to a higher level of rne mRNA and a lower level of sinI mRNA independently of the presence of ExpR, the AHL receptor, and AHLs. In line with this, IPTG (isopropyl-β-D-thiogalactopyranoside)-induced overexpression of rne resulted in a shorter half-life of sinI mRNA and a strong reduction of AHL accumulation. Moreover, using translational sinI-egfp fusions, we found that sinI expression is specifically decreased upon induced overexpression of rne, independently of the presence of the global posttranscriptional regulator Hfq. The 28-nucleotide 5' untranslated region (UTR) of sinI mRNA was sufficient for this effect. Random amplification of 5' cDNA ends (5'-RACE) analyses revealed a potential RNase E cleavage site at position +24 between the Shine-Dalgarno site and the translation start site. We postulate therefore that RNase E-dependent degradation of sinI mRNA from the 5' end is one of the steps mediating a high turnover of sinI mRNA, which allows the Sin quorum-sensing system to respond rapidly to changes in transcriptional control of AHL production.
苜蓿中华根瘤菌的群体感应依赖 N-酰基高丝氨酸内酯 (AHL) 作为自体诱导物。在高密度时,AHL 的产生会增加,这取决于 AHL 合酶 SinI 和两个转录调节因子 SinR 和 ExpR。我们的研究表明,编码内切核糖核酸酶 RNase E 的 rne 基因的异位表达会阻止 AHL 积累到可检测水平。rne 基因的异位表达导致 rne mRNA 的水平升高,而 sinI mRNA 的水平降低,而 ExpR、AHL 受体和 AHLs 的存在与否均不受影响。与此一致的是,IPTG(异丙基-β-D-硫代半乳糖吡喃糖苷)诱导的 rne 过表达导致 sinI mRNA 的半衰期缩短,AHL 积累量大幅减少。此外,通过翻译 sinI-egfp 融合,我们发现 rne 的过表达会特异性地降低 sinI 的表达,而与全局转录后调节因子 Hfq 的存在与否无关。sinI mRNA 的 28 个核苷酸 5'非翻译区(UTR)足以产生这种效应。随机扩增的 5'cDNA 末端(5'-RACE)分析显示,在 Shine-Dalgarno 位点和翻译起始位点之间的+24 位置可能存在 RNase E 切割位点。因此,我们假设 RNase E 依赖的 sinI mRNA 从 5'端的降解是介导 sinI mRNA 高周转率的步骤之一,这使得 Sin 群体感应系统能够快速响应 AHL 产生的转录控制变化。