Department of Microbiology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
J Bacteriol. 2012 Oct;194(19):5228-36. doi: 10.1128/JB.00941-12. Epub 2012 Jul 20.
CTnDOT is a 65-kb conjugative transposon present in Bacteroides spp. that confers resistance to erythromycin [erm(F)] and tetracycline [tet(Q)]. An interesting feature of CTnDOT is that both excision from the chromosome and transfer of CTnDOT are stimulated by exposure to tetracycline. However, when no tetracycline is present, transfer of CTnDOT is not detectable. Previous studies suggested that a region containing a small RNA, RteR, appeared to mediate repression of CTnDOT transfer; however, virtually nothing was known about RteR. We have demonstrated that RteR is a 90-nucleotide transcript that is not further processed. RteR inhibits conjugative transfer of CTnDOT by targeting the transfer region, a 13-kb operon that encodes the tra genes required to assemble the mating apparatus. We report here that RteR interacts with the region downstream of traA. Levels of the downstream tra mRNA are dramatically reduced when RteR is present. Further, RteR does not appear to decrease the half-life of the tra mRNA transcript, suggesting that RteR does not bind to the transcript to initiate RNase-dependent decay, similar to other trans-acting small RNAs. We predict that RteR may act to enhance termination of the tra operon within traB, which could account for the decreased abundance of the tra transcript downstream of traA and explain why the tra mRNA has the same half-life whether or not RteR is present. RteR is the only small RNA that has been characterized so far within the Bacteroidetes phylum.
CTnDOT 是一种存在于拟杆菌属中的 65kb 可接合转座子,可赋予红霉素 [erm(F)] 和四环素 [tet(Q)] 的抗性。CTnDOT 的一个有趣特征是,无论是从染色体上切除还是转移 CTnDOT,都会受到四环素的刺激。然而,当没有四环素存在时,就无法检测到 CTnDOT 的转移。先前的研究表明,一个包含小 RNA RteR 的区域似乎介导了 CTnDOT 转移的抑制;然而,人们对 RteR 几乎一无所知。我们已经证明,RteR 是一个 90 个核苷酸的转录本,不会进一步加工。RteR 通过靶向转移区域来抑制 CTnDOT 的接合转移,转移区域是一个编码组装交配装置所需的 tra 基因的 13kb 操纵子。我们在这里报告 RteR 与 traA 下游区域相互作用。当存在 RteR 时,下游 tra mRNA 的水平会显著降低。此外,RteR 似乎不会降低 tra mRNA 转录本的半衰期,这表明 RteR 不会结合转录本以启动 RNase 依赖性衰减,这与其他反式作用小 RNA 相似。我们预测 RteR 可能会促进 tra 操纵子在 traB 内的终止,这可以解释为什么 traA 下游的 tra 转录本丰度降低,以及为什么无论是否存在 RteR,tra mRNA 的半衰期都相同。RteR 是迄今为止在拟杆菌门中唯一被表征的小 RNA。