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Functional Characterization of EpsC a Component of the Type II Secretion System in the Pathogenicity of Vibrio vulnificus

机译:EpsCII型分泌系统的组成部分在创伤弧菌致病性中的功能表征

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摘要

EpsC, one of the components comprising the type II secretion system (T2SS), was isolated from a human-pathogenic bacterium, Vibrio vulnificus, to evaluate its role in eliciting virulence. An espC-deleted mutant of V. vulnificus displayed a reduced cytotoxicity to the human cell line HEp-2 and an attenuated virulence in a mouse model. This mutant exhibited dramatic defects in the secretion of diverse extracellular proteins, such as outer membrane proteins, transporters, and the known secreted factors, notably, a hemolysin (VvhA) and an elastase (VvpE). A defect in its secretion of proteins was restored by in trans complementation of the intact epsC gene. Analyses of cellular fractions revealed that VvhA and VvpE of the ΔepsC mutant were not excreted outside the cell but were present mainly in the periplasmic space. Examination of a V. vulnificus mutant deficient in TolC, a component of the T1SS, showed that it is not involved in the secretion of VvhA and VvpE but that it is necessary for the secretion of another major toxin of V. vulnificus, RtxA. Therefore, the T2SS is required for V. vulnificus pathogenicity, which is mediated by at least two secreted factors, VvhA and VvpE, via facilitating the secretion and exposure of these factors to host cells.
机译:从人类致病性细菌弧菌弧菌中分离出EpsC,它是构成II型分泌系统(T2SS)的成分之一,以评估其在诱发毒力中的作用。 espC缺失的V. vulnificus突变体在小鼠模型中显示出对人细胞系HEp-2的细胞毒性降低和毒力减弱。该突变体在多种细胞外蛋白(例如外膜蛋白,转运蛋白)和已知的分泌因子(尤其是溶血素(VvhA)和弹性蛋白酶(VvpE))的分泌中表现出明显的缺陷。通过完整的epsC基因的反式互补可以恢复蛋白质分泌的缺陷。细胞成分分析表明,ΔepsC突变体的VvhA和VvpE并未排泄到细胞外,而是主要存在于周质空间。对TolC(T1SS的一个组成部分)缺乏的V. vulnificus突变体的检查表明,它不参与VvhA和VvpE的分泌,但是对于V.vulnificus的另一种主要毒素RtxA的分泌是必需的。因此,T2SS是创伤弧菌致病性所必需的,它通过促进这些因子向宿主细胞的分泌和暴露,由至少两个分泌因子VvhA和VvpE介导。

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