Ellis Michael J, Trussler Ryan S, Charles Onella, Haniford David B
Department of Biochemistry, University of Western Ontario, London, Ontario N6A 5C1, Canada.
Nucleic Acids Res. 2017 May 19;45(9):5470-5486. doi: 10.1093/nar/gkx094.
Bacterial sRNAs play an important role in regulating many cellular processes including metabolism, outer membrane homeostasis and virulence. Although sRNAs were initially found in intergenic regions, there is emerging evidence that protein coding regions of the genome are a rich reservoir of sRNAs. Here we report that the 5΄UTR of IS200 transposase mRNA (tnpA) is processed to produce regulatory RNAs that affect expression of over 70 genes in Salmonella Typhimurium. We provide evidence that the tnpA derived sRNA base-pairs with invF mRNA to repress expression. As InvF is a transcriptional activator of SPI-1 encoded and other effector proteins, tnpA indirectly represses these genes. We show that deletion of IS200 elements in S. Typhimurium increases invasion in vitro and reduces growth rate, while over-expression of tnpA suppresses invasion. Our work indicates that tnpA acts as an sRNA 'sponge' that sets a threshold for activation of Salmonella pathogenicity island (SPI)-1 effector proteins and identifies a new class of 'passenger gene' for bacterial transposons, providing the first example of a bacterial transposon producing a regulatory RNA that controls host gene expression.
细菌小RNA在调控包括代谢、外膜稳态和毒力在内的许多细胞过程中发挥着重要作用。尽管小RNA最初是在基因间区域发现的,但越来越多的证据表明,基因组的蛋白质编码区域是小RNA的丰富来源。在此,我们报道IS200转座酶mRNA(tnpA)的5΄非翻译区(UTR)被加工产生调控RNA,这些RNA影响鼠伤寒沙门氏菌中70多个基因的表达。我们提供的证据表明,tnpA衍生的小RNA与invF mRNA碱基配对以抑制表达。由于InvF是SPI-1编码的其他效应蛋白的转录激活因子,tnpA间接抑制这些基因。我们表明,删除鼠伤寒沙门氏菌中的IS200元件会增加体外侵袭并降低生长速率,而tnpA的过表达则抑制侵袭。我们的工作表明,tnpA作为一种小RNA“海绵”,为沙门氏菌致病岛(SPI)-1效应蛋白的激活设定了阈值,并确定了一类新的细菌转座子“乘客基因”,这是细菌转座子产生控制宿主基因表达的调控RNA的首个例子。