The (p)ppGpp synthetase Rsh promotes rifampicin tolerant persister cell formation in Brucella abortus by regulating the type II toxin-antitoxin module mbcTA

Xiaofan Liu, Pingping Wang, Ningqiu Yuan, Yunyi Zhai, Yuanhao Yang, Mingyue Hao, Mingxing Zhang, Dong Zhou, Wei Liu, Yaping Jin, Aihua Wang
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Abstract

Persister cells are transiently tolerant to antibiotics and are associated with recalcitrant chronic infections due to recolonization of host cells after antibiotic removal. Brucella spp. are facultative pathogens that establish intracellular infection cycles in host cells which results in chronic persistent infections. Brucella abortus forms multi-drug persister cells which are promoted by the (p)ppGpp synthetase Rsh during rifampicin exposure. Here, we confirmed that Rsh promoted persister cells formation in B. abortus stationary phase treated with rifampicin and enrofloxacin. Deletion of the gene for Rsh decreased persister cells level in the presence of these drugs in different growth phases. However, persister cells formation by deletion strain varied in different growth phases in the presence of other antibiotics. Rsh also was involved in persister cells formation during rifampicin treatment under certain stress conditions, including acidic conditions, exposure to PBS, and heat stress. Moreover, Rsh impacted persister cell levels during rifampicin or enrofloxacin treatment in RAW264.7 macrophages. Certain typeIItoxin-antitoxin modules were upregulated under various stress conditions in B. abortus. We established that Rsh positively regulated the type II toxin-antitoxin mbcTA. Moreover, rifampicin-tolerant persister cells formation was elevated and ATP levels were decreased when mbcTA promoter was overexpressed in Rsh deletion background in stationary phase. Our results establish that (p)ppGpp synthetase Rsh plays a key role in B. abortus persistence and may serve as a potent novel target in combination with rifampicin in the development of new therapeutic approaches and prevention strategies to treat chronic infections of Brucella.
(pp)ppGpp合成酶Rsh通过调节II型毒素-抗毒素模块mbcTA,促进流产布鲁氏菌中利福平耐受性宿主细胞的形成
顽固细胞对抗生素具有短暂耐受性,由于抗生素去除后宿主细胞会重新定殖,因此会出现顽固性慢性感染。布鲁氏菌属是面性病原体,可在宿主细胞内建立细胞内感染循环,导致慢性持续感染。流产布鲁氏菌会形成多药宿主细胞,在利福平暴露期间,(pp)ppGpp合成酶Rsh会促进这种宿主细胞的形成。在这里,我们证实 Rsh 能促进用利福平和恩诺沙星处理的流产布鲁氏菌静止期顽固细胞的形成。缺失 Rsh 基因会降低这些药物作用下不同生长阶段的宿主细胞水平。然而,在有其他抗生素存在的不同生长阶段,缺失基因的菌株形成的宿主细胞也不同。在某些胁迫条件下,包括酸性条件、暴露于 PBS 和热胁迫,Rsh 也参与了利福平处理过程中的固着细胞形成。此外,Rsh 还影响 RAW264.7 巨噬细胞在利福平或恩诺沙星处理期间的固着细胞水平。在各种应激条件下,某些II型毒素-抗毒素模块在流产杆菌中上调。我们发现,Rsh 对 II 型毒素-抗毒素 mbcTA 具有正向调节作用。此外,在Rsh缺失背景下,当mbcTA启动子在静止期过度表达时,利福平耐受性顽固细胞的形成增加,ATP水平降低。我们的研究结果表明,(p)ppGpp 合成酶 Rsh 在布鲁氏菌的宿存过程中起着关键作用,与利福平联用可作为一个有效的新靶点,用于开发治疗布鲁氏菌慢性感染的新疗法和预防策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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