Yersinia pseudotuberculosis doxycycline tolerance strategies include modulating expression of genes involved in cell permeability and tRNA modifications.
Yersinia pseudotuberculosis doxycycline tolerance strategies include modulating expression of genes involved in cell permeability and tRNA modifications.
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DOI：
10.1371/journal.ppat.1010556
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发表时间：
2022-05
期刊：
影响因子：
6.7
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中科院分区：
文献类型：
--
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抗生素耐受性通常与细菌群体的表型变化有关，导致抗生素敏感性的短暂降低，从而导致治疗失败和复发性感染。虽然耐受细胞可能在治疗前出现，但长期暴露于抗生素的压力也可以促进耐受性。在这里，我们试图确定假结核耶尔森菌对强力霉素暴露的反应，然后验证这些基因表达变化是否可以促进培养和我们的小鼠感染模型中的强力霉素耐受性。在生理相关剂量的强力霉素作用下，只有4个基因发生了差异调控：osmB和ompF上调，tusB和cnfy下调；在多西环素治疗期间，小鼠也出现了差异表达。ompF、tusB和cnfy对氯霉素的反应也有差异调节，表明这些可能是对核糖体抑制的一般反应。以前，Cnfy与持久性有关，并不是本文的主要关注点。我们发现OmpF孔蛋白的缺失导致抗生素积累增加，这表明表达可能促进强力霉素向细胞外扩散，而OmpF脂蛋白对抗生素的渗透性影响较小。tusB的过表达显著损害了细菌在培养物和小鼠中的存活，这表明tusB修饰tRNA及其对翻译机制的影响，促进了传统上被认为是抑菌剂的抗生素治疗期间的存活。我们认为这可能是首次观察到强力霉素在生理条件下的杀菌活性，这是通过逆转tusB下调而揭示的。本研究的最初目的是确定假结核耶尔森菌如何在多西环素暴露下发生转录变化，然后确定这些转录变化是否需要在长时间的药物暴露下存活。我们的数据表明，孔蛋白的表达不仅参与抗生素进入细菌细胞，还可能促进强力霉素在细胞外的被动扩散。出乎意料的是，我们还发现tusB（一种参与tRNA修饰的基因产物）的过表达导致强力霉素在典型的最低抑制浓度下对假结核杆菌具有有效的杀菌活性。强力霉素通常被认为是一种抑菌抗生素，它可以可逆地结合核糖体，从而抑制细菌的生长。然而，越来越多的证据表明抑菌抗生素也具有杀菌活性。我们相信我们的结果可能是第一次证明强力霉素在生理条件下的杀菌活性；一种细菌病原体，其药物浓度可在小鼠和人体组织中获得。
Antibiotic tolerance is typically associated with a phenotypic change within a bacterial population, resulting in a transient decrease in antibiotic susceptibility that can contribute to treatment failure and recurrent infections. Although tolerant cells may emerge prior to treatment, the stress of prolonged antibiotic exposure can also promote tolerance. Here, we sought to determine how Yersinia pseudotuberculosis responds to doxycycline exposure, to then verify if these gene expression changes could promote doxycycline tolerance in culture and in our mouse model of infection. Only four genes were differentially regulated in response to a physiologically-relevant dose of doxycycline: osmB and ompF were upregulated, tusB and cnfy were downregulated; differential expression also occurred during doxycycline treatment in the mouse. ompF, tusB and cnfy were also differentially regulated in response to chloramphenicol, indicating these could be general responses to ribosomal inhibition. cnfy has previously been associated with persistence and was not a major focus here. We found deletion of the OmpF porin resulted in increased antibiotic accumulation, suggesting expression may promote diffusion of doxycycline out of the cell, while OsmB lipoprotein had a minor impact on antibiotic permeability. Overexpression of tusB significantly impaired bacterial survival in culture and in the mouse, suggesting that tRNA modification by tusB, and the resulting impacts on translational machinery, promotes survival during treatment with an antibiotic classically viewed as bacteriostatic. We believe this may be the first observation of bactericidal activity of doxycycline under physiological conditions, which was revealed by reversing tusB downregulation. The initial goal of this study was to determine how Yersinia pseudotuberculosis changes transcriptionally in response to doxycycline exposure, to then determine if these transcriptional changes are required to survive prolonged drug exposure. Our data suggests that porin expression is not just involved in antibiotic entry into bacterial cells, and may also promote passive diffusion of doxycycline out of the cell. Unexpectedly, we also found that overexpression of tusB, a gene product involved in tRNA modifications, resulted in potent doxycycline bactericidal activity at the typical minimum inhibitory concentration for Y. pseudotuberculosis. Doxycycline is classically viewed as a bacteriostatic antibiotic, which reversibly binds ribosomes, resulting in bacterial growth inhibition. However, growing evidence suggests that bacteriostatic antibiotics can also have bactericidal activity. We believe our results may be the first demonstration of the bactericidal activity of doxycycline under physiological conditions; with a bacterial pathogen, at a drug concentration that can be obtained within mouse and human tissues.
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影响因子：
5.2
作者：
HOOGKAMPKORSTANJE, JAA
通讯作者：
HOOGKAMPKORSTANJE, JAA
影响因子：
3.1
作者：
Davis, Kimberly M.;Krupp, Joanna;Isberg, Ralph R.
通讯作者：
Isberg, Ralph R.
影响因子：
4.9
作者：
BONACORSI, SP;SCAVIZZI, MR;CARNIEL, E
通讯作者：
CARNIEL, E
影响因子：
6.7
作者：
Crimmins, Gregory T.;Mohammadi, Sina;Mecsas, Joan
通讯作者：
Mecsas, Joan
DOI：
10.1073/pnas.1316084111
发表时间：
2014-01-07
影响因子：
11.1
作者：
Antonia Sanchez-Romero, Maria;Casadesus, Josep
通讯作者：
Casadesus, Josep