Imamovic Lejla, Martínez-Castillo Alexandre, Benavides Carmen, Muniesa Maite
Department of Microbiology, University of Barcelona, Barcelona, Spain.
Department of Microbiology, University of Barcelona, Barcelona, Spain
Infect Immun. 2015 Apr;83(4):1451-7. doi: 10.1128/IAI.02916-14. Epub 2015 Jan 26.
Infection and lysogenic conversion with Shiga toxin-encoding bacteriophages (Stx phages) drive the emergence of new Shiga toxin-producing Escherichia coli strains. Phage attachment to the bacterial surface is the first stage of phage infection. Envelope perturbation causes activation of envelope stress responses in bacterial cells. Although many external factors are known to activate envelope stress responses, the role of these responses in the phage-bacterium interaction remains unexplored. Here, we investigate the link between three envelope signaling systems in E. coli (RcsBC, CpxAR, and BaeSR) and Stx2 phage infection by determining the success of bacterial lysogenic conversion. For this purpose, E. coli DH5α wild-type (WT) and mutant strains lacking RcsBC, CpxAR, or BaeSR signaling systems were incubated with a recombinant Stx2 phage (933W). Notably, the number of lysogens obtained with the BaeSR mutant was 5 log10 units higher than with the WT, and the same differences were observed when using 7 different Stx2 phages. To assess whether the membrane receptor used by Stx phages, BamA, was involved in the differences observed, bamA gene expression was monitored by reverse transcription-quantitative PCR (RT-qPCR) in all host strains. A 4-fold-higher bamA expression level was observed in the BaeSR mutant than in the WT strain, suggesting that differential expression of the receptor used by Stx phages accounted for the increase in the number of lysogenization events. Establishing the link between the role of stress responses and phage infection has important implications for understanding the factors affecting lysogenic conversion, which drives the emergence of new pathogenic clones.
感染并携带编码志贺毒素的噬菌体(Stx噬菌体)进行溶原性转化会促使新的产志贺毒素大肠杆菌菌株出现。噬菌体附着于细菌表面是噬菌体感染的第一阶段。包膜扰动会激活细菌细胞中的包膜应激反应。尽管已知许多外部因素可激活包膜应激反应,但这些反应在噬菌体 - 细菌相互作用中的作用仍未得到探索。在此,我们通过确定细菌溶原性转化的成功率,研究了大肠杆菌中的三种包膜信号系统(RcsBC、CpxAR和BaeSR)与Stx2噬菌体感染之间的联系。为此,将大肠杆菌DH5α野生型(WT)以及缺乏RcsBC、CpxAR或BaeSR信号系统的突变菌株与重组Stx2噬菌体(933W)一起培养。值得注意的是,用BaeSR突变体获得的溶原菌数量比野生型高5个对数10单位,并且在使用7种不同的Stx2噬菌体时也观察到了相同的差异。为了评估Stx噬菌体使用的膜受体BamA是否参与了观察到的差异,通过逆转录定量PCR(RT - qPCR)监测了所有宿主菌株中bamA基因的表达。在BaeSR突变体中观察到的bamA表达水平比野生型菌株高4倍,这表明Stx噬菌体使用的受体的差异表达导致了溶原化事件数量的增加。确定应激反应的作用与噬菌体感染之间的联系对于理解影响溶原性转化的因素具有重要意义,溶原性转化会促使新的致病克隆出现。