Comprehensive identification of Vibrio vulnificus genes required for growth in human serum.
Comprehensive identification of Vibrio vulnificus genes required for growth in human serum.
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DOI：
10.1080/21505594.2018.1455464
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发表时间：
2018-12-31
期刊：
影响因子：
5.2
通讯作者：
Amaro C
中科院分区：
文献类型：
--
作者：
Carda-Diéguez M;Silva-Hernández FX;Hubbard TP;Chao MC;Waldor MK;Amaro C
创伤弧菌是一种高度侵入性的病原体，能够从感染部位传播到血液，导致败血症和死亡。为了在血液中生存和增殖，病原体需要克服这种宿主生态位的先天免疫防御和代谢限制的机制。我们在YJ 016中创建了一个高密度转座子突变体文库，YJ 016是最毒力创伤弧菌谱系（或V. vulnificus组）的代表菌株，并使用转座子插入测序（TIS）筛选来鉴定使病原体能够在人血清中存活和增殖的基因座。最初，用于插入的代表性不足的基因用于估计创伤弧菌必需基因集;将这些基因与其他弧菌中代表性不足的基因的类似基于TIS的分类进行比较，使得能够编译共同的弧菌必需基因集。在暴露于血清后，对文库中插入突变体的相对丰度的分析表明，参与荚膜生物发生的基因对于YJ 016补体抗性至关重要。值得注意的是，YJ 016血清抗性和荚膜生物发生所需的两个基因的同源物先前与荚膜生物发生无关，并且在其他创伤弧菌菌株中基本上不存在。将暴露于热灭活血清后突变体的相对丰度与血清筛选的结果进行比较。这些比较表明，在这两种条件下，病原体依赖于其Na+转运NADH-泛醌还原酶（NQR）复合物和II型分泌系统生存/增殖的代谢限制内的血清。总的来说，我们的研究结果揭示了比较TIS筛选的效力，以提供病原体如何克服血清对生长施加的各种限制的知识。
Vibrio vulnificus can be a highly invasive pathogen capable of spreading from an infection site to the bloodstream, causing sepsis and death. To survive and proliferate in blood, the pathogen requires mechanisms to overcome the innate immune defenses and metabolic limitations of this host niche. We created a high-density transposon mutant library in YJ016, a strain representative of the most virulent V. vulnificus lineage (or phylogroup) and used transposon insertion sequencing (TIS) screens to identify loci that enable the pathogen to survive and proliferate in human serum. Initially, genes underrepresented for insertions were used to estimate the V. vulnificus essential gene set; comparisons of these genes with similar TIS-based classification of underrepresented genes in other vibrios enabled the compilation of a common Vibrio essential gene set. Analysis of the relative abundance of insertion mutants in the library after exposure to serum suggested that genes involved in capsule biogenesis are critical for YJ016 complement resistance. Notably, homologues of two genes required for YJ016 serum-resistance and capsule biogenesis were not previously linked to capsule biogenesis and are largely absent from other V. vulnificus strains. The relative abundance of mutants after exposure to heat inactivated serum was compared with the findings from the serum screen. These comparisons suggest that in both conditions the pathogen relies on its Na+ transporting NADH-ubiquinone reductase (NQR) complex and type II secretion system to survive/proliferate within the metabolic constraints of serum. Collectively, our findings reveal the potency of comparative TIS screens to provide knowledge of how a pathogen overcomes the diverse limitations to growth imposed by serum.
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