Zhong Qiu, Zhao Yan, Chen Tian, Yin Supeng, Yao Xinyue, Wang Jing, Lu Shuguang, Tan Yinling, Tang Jiaqi, Zheng Beiwen, Hu Fuquan, Li Ming
Department of Microbiology, Third Military Medical University, Chongqing 400038, China.
BMC Microbiol. 2014 Mar 22;14:73. doi: 10.1186/1471-2180-14-73.
Streptococcus suis serotype 2 (S. suis 2) has evolved efficient mechanisms to cause streptococcal toxic shock syndrome (STSS), which is a new emerging infectious disease linked to S. suis. We have previously reported that a type IV secretion system (T4SS) harbored by the specific 89K pathogenicity island (PAI) of S. suis 2 contributes to the development of STSS and mediates horizontal transfer of 89K. However, the 89K T4SS machinery assembly in vivo and in vitro is poorly understood, and the component acting directly to digest the bacterial cell wall needs to be identified.
The virB1-89K gene product encoded in the 89K PAI is the only one that shows similarity to the Agrobacterium VirB1 component and contains a conserved CHAP domain that may function in peptidoglycan hydrolysis, which makes it a plausible candidate acting as a hydrolase against the peptidoglycan cell wall to allow the assembly of the T4SS apparatus. In the current study, the CHAP domain of VirB1-89K from S. suis 89K PAI was cloned and over-expressed in Escherichia coli, and its peptidoglycan-degrading activity in vitro was determined. The results indicated that the VirB1-89K CHAP domain can degrade the peptidoglycan layer of bacteria. Deletion of virB1-89K reduces significantly, but does not abolish, the virulence of S. suis in a mouse model.
The experimental results presented here suggested that VirB1-89K facilitates the assembly of 89K T4SS apparatus by catalyzing the degradation of the peptidoglycan cell wall, thus contributing to the pathogenesis of S. suis 2 infection.
猪链球菌2型(S. suis 2)已进化出有效的机制来引发链球菌中毒性休克综合征(STSS),这是一种与猪链球菌相关的新出现的传染病。我们之前报道过,猪链球菌2型特定的89K致病岛(PAI)所携带的IV型分泌系统(T4SS)有助于STSS的发展并介导89K的水平转移。然而,人们对89K T4SS在体内和体外的组装机制了解甚少,且需要鉴定直接作用于消化细菌细胞壁的成分。
89K PAI中编码的virB1 - 89K基因产物是唯一与根癌农杆菌VirB1成分具有相似性的产物,并且包含一个保守的CHAP结构域,该结构域可能在肽聚糖水解中发挥作用,这使其成为作为针对肽聚糖细胞壁的水解酶以允许T4SS装置组装的合理候选物。在本研究中,克隆了来自猪链球菌89K PAI的VirB1 - 89K的CHAP结构域并在大肠杆菌中进行了过表达,并测定了其体外肽聚糖降解活性。结果表明,VirB1 - 89K CHAP结构域可以降解细菌的肽聚糖层。在小鼠模型中,缺失virB1 - 89K会显著降低但不会消除猪链球菌的毒力。
此处呈现的实验结果表明VirB1 - 89K通过催化肽聚糖细胞壁的降解促进89K T4SS装置的组装,从而有助于猪链球菌2型感染的发病机制。