Center for Microbial Dynamics and Infection, Georgia Institute of Technology, 310 Ferst Drive, Atlanta, GA, 30332-0230, USA.
Institute for Bioengineering and Biosciences, Georgia Institute of Technology, 310 Ferst Drive, Atlanta, GA, 30332-0230, USA.
Genome Biol. 2019 Aug 12;20(1):163. doi: 10.1186/s13059-019-1765-5.
Like many bacteria, Vibrio cholerae deploys a harpoon-like type VI secretion system (T6SS) to compete against other microbes in environmental and host settings. The T6SS punctures adjacent cells and delivers toxic effector proteins that are harmless to bacteria carrying cognate immunity factors. Only four effector/immunity pairs encoded on one large and three auxiliary gene clusters have been characterized from largely clonal, patient-derived strains of V. cholerae.
We sequence two dozen V. cholerae strain genomes from diverse sources and develop a novel and adaptable bioinformatics tool based on hidden Markov models. We identify two new T6SS auxiliary gene clusters and describe Aux 5 here. Four Aux 5 loci are present in the host strain, each with an atypical effector/immunity gene organization. Structural prediction of the putative effector indicates it is a lipase, which we name TleV1 (type VI lipase effector Vibrio). Ectopic TleV1 expression induces toxicity in Escherichia coli, which is rescued by co-expression of the TliV1a immunity factor. A clinical V. cholerae reference strain expressing the Aux 5 cluster uses TleV1 to lyse its parental strain upon contact via its T6SS but is unable to kill parental cells expressing the TliV1a immunity factor.
We develop a novel bioinformatics method and identify new T6SS gene clusters in V. cholerae. We also show the TleV1 toxin is delivered in a T6SS manner by V. cholerae and can lyse other bacterial cells. Our web-based tool can be modified to identify additional novel T6SS genomic loci in diverse bacterial species.
与许多细菌一样,霍乱弧菌利用一种类似鱼叉的 VI 型分泌系统(T6SS)在环境和宿主环境中与其他微生物竞争。T6SS 刺穿相邻的细胞,并输送对携带同源免疫因子的细菌无害的毒性效应蛋白。仅从主要是克隆的、源自患者的霍乱弧菌菌株中,就已经鉴定出四个效应物/免疫对,这些效应物/免疫对编码在一个大型和三个辅助基因簇上。
我们对来自不同来源的二十多个霍乱弧菌菌株进行了测序,并基于隐马尔可夫模型开发了一种新颖且适应性强的生物信息学工具。我们鉴定出两个新的 T6SS 辅助基因簇,并在这里描述 Aux5。宿主菌株中有四个 Aux5 基因座,每个基因座都具有非典型的效应物/免疫基因组织。推测的效应物的结构预测表明它是一种脂肪酶,我们将其命名为 TleV1(VI 型脂肪酶效应物霍乱弧菌)。异位表达 TleV1 会在大肠杆菌中诱导毒性,而 TliV1a 免疫因子的共表达可以挽救毒性。表达 Aux5 簇的临床霍乱弧菌参考菌株通过其 T6SS 接触时会利用 TleV1 裂解其亲本菌株,但无法杀死表达 TliV1a 免疫因子的亲本细胞。
我们开发了一种新的生物信息学方法,并在霍乱弧菌中鉴定了新的 T6SS 基因簇。我们还表明,TleV1 毒素通过霍乱弧菌以 T6SS 的方式传递,并可以裂解其他细菌细胞。我们的基于网络的工具可以修改,以识别其他细菌物种中更多的新型 T6SS 基因组基因座。