Investigating the effect of bacteriophages on bacterial FtsZ localisation.
Investigating the effect of bacteriophages on bacterial FtsZ localisation.
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DOI：
10.3389/fcimb.2022.863712
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发表时间：
2022
影响因子：
5.7
通讯作者：
中科院分区：
文献类型：
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大肠杆菌是最常见的革兰氏阴性病原体之一，是导致新生儿脑膜炎和败血症的感染的原因。FtsZ蛋白是大多数物种（包括大肠杆菌）细胞分裂所需的细菌微管蛋白同源物。杆菌几种阻断细胞分裂的试剂已显示出错误定位FtsZ，包括噬菌体λ编码的Kil肽，导致有缺陷的细胞分裂和丝状表型，使FtsZ成为抗微生物剂的有吸引力的靶标。在这项研究中，我们使用了一个体外脑膜炎模型系统，研究了噬菌体对FtsZ的影响，使用荧光E。coli EV 36/FtsZ-mCherry和K12/FtsZ-mNeon菌株。我们显示在正常生长条件下FtsZ作为单环定位于细菌细胞中间体，并且在添加已知抑制剂Kil肽后产生丝状多环细菌细胞的FtsZ的错误定位。我们还表明，当噬菌体K1 F-GFP和T7-mCherry应用于其各自的宿主菌株时，这些噬菌体可以抑制FtsZ并阻断细菌细胞分裂，导致丝状多环表型，潜在地延迟裂解并增加后代数量。这发生在指数增长阶段，因为需要活跃的分裂宿主。我们提出，ZapA蛋白是需要通过显示ZapA突变株的表型恢复的噬菌体抑制，我们表明，FtsI蛋白也是错误定位的噬菌体感染后。最后，我们通过观察T7Δ0.4突变体的表型恢复，表明T7肽gp0.4负责K12菌株中FtsZ的抑制。
Escherichia coli is one of the most common Gram-negative pathogens and is responsible for infection leading to neonatal meningitis and sepsis. The FtsZ protein is a bacterial tubulin homolog required for cell division in most species, including E. coli. Several agents that block cell division have been shown to mislocalise FtsZ, including the bacteriophage λ-encoded Kil peptide, resulting in defective cell division and a filamentous phenotype, making FtsZ an attractive target for antimicrobials. In this study, we have used an in vitro meningitis model system for studying the effect of bacteriophages on FtsZ using fluorescent E. coli EV36/FtsZ-mCherry and K12/FtsZ-mNeon strains. We show localisation of FtsZ to the bacterial cell midbody as a single ring during normal growth conditions, and mislocalisation of FtsZ producing filamentous multi-ringed bacterial cells upon addition of the known inhibitor Kil peptide. We also show that when bacteriophages K1F-GFP and T7-mCherry were applied to their respective host strains, these phages can inhibit FtsZ and block bacterial cell division leading to a filamentous multi-ringed phenotype, potentially delaying lysis and increasing progeny number. This occurs in the exponential growth phase, as actively dividing hosts are needed. We present that ZapA protein is needed for phage inhibition by showing a phenotype recovery with a ZapA mutant strain, and we show that FtsI protein is also mislocalised upon phage infection. Finally, we show that the T7 peptide gp0.4 is responsible for the inhibition of FtsZ in K12 strains by observing a phenotype recovery with a T7Δ0.4 mutant.