Roles of virulence regulator ToxR in viable but non-culturable formation by controlling reactive oxygen species resistance in pathogen Vibrio alginolyticus.
Roles of virulence regulator ToxR in viable but non-culturable formation by controlling reactive oxygen species resistance in pathogen Vibrio alginolyticus.
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毒力调节剂 ToxR 通过控制病原体溶藻弧菌中的活性氧抗性在活的但不可培养的形成中的作用。
DOI：
10.1016/j.micres.2021.126900
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发表时间：
2021-10
影响因子：
6.7
通讯作者：
Shuai Shao
中科院分区：
文献类型：
--
作者：
Mengqing Zhou;Yuping Huang;Yuanxing Zhang;Qiyao Wang;Yue Ma;Shuai Shao
在不利的环境下，细菌进入活的但不可培养（VBNC）状态，一种类似休眠的生存状态。改变的基因调控基础的VBNC状态的进入尚未得到很好的阐明。在这里，我们报道了一个亚群的细胞（23.8%）inVibrio alginolyticuscultures进入VBNC状态响应营养限制在碱性pH值。在这些条件下的关键毒力调节因子ToxR的蛋白水解与VBNC的形成。同时，ToxR基因的缺失削弱了毒力相关基因的表达和迁移能力，导致病毒毒力减弱。溶藻菌在VBNC诱导条件下生长的细胞的RNA-seq和ChIP-seq分析显示，ToxR直接控制包括参与活性氧（ROS）抗性的ahp和Canddps在内的18个基因的表达。ToxR与kdgR、ppiC、ahpC和dps的启动子区结合，并进一步控制它们在氧化应激条件下的各自表达。ToxR受损的细胞积累有害的细胞内ROS。此外，这些基因有助于细菌的可培养性，因为它们的框内缺失菌株表现出严重降低的平板计数，而互补菌株显示出挽救的活力。总的来说，这项研究揭示了ToxR的作用，在开关上的VBNC状态通过感应不利的环境信号，如内源性ROS（过氧化氢，H2 O2）inV。alginolyticus和提供了机械的见解tobriolifestyle适应海洋环境。
Under adverse circumstances, bacteria enter the viable but non-culturable (VBNC) state, a dormancy-like state for survival. The altered gene regulation underlying the entry of the VBNC state has not yet been well elucidated. Here, we reported that a subpopulation of cells (23.8 %) inVibrio alginolyticuscultures enters the VBNC state in response to nutrient limitation at alkaline pH. The proteolysis of pivotal virulence regulator ToxR at these conditions is associated with VBNC formation. Meantime, ToxR abrogation impaired the mobility and the expression of virulence-associated genes, resulting in attenuated virulence inV. alginolyticus. RNA-seq and ChIP-seq analyses of the cells grown in VBNC-inducing conditions revealed that ToxR directly controls the expression of ∼8 genes includingahpCanddps involved in reactive oxygen species (ROS) resistance. ToxR binds to the promoter regions ofkdgR, ppiC, ahpC,anddps and further controls their respective expression under oxidative stress conditions. The cells with impaired ToxR accumulated detrimental intracellular ROS. Moreover, these genes contribute to bacterial culturability as their in-frame deletion strains exhibiting severely decreased plate counts and the complementary strain showed rescued viability. Collectively, this study revealed the role of ToxR in switching on the VBNC state by sensing unfavorable environmental signals such as endogenous ROS (hydrogen peroxide, H2O2) inV. alginolyticusand provided mechanistic insights intoVibriolifestyle adaptation in the marine environment.
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