Zhang J, Xiao J, Zhang Y, Cui S, Liu Q, Wang Q, Wu H, Zhang Y
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
Lett Appl Microbiol. 2014 Nov;59(5):557-64. doi: 10.1111/lam.12316. Epub 2014 Sep 12.
The evpP gene in fish pathogen Edwardsiella tarda, coding the T6SS secretory protein EvpP and carrying an evpA-evpO independent promoter region, was crucial for host cell invasion. The transcription of evpP was positively regulated by either the two-component system EsrA-EsrB or iron concentration, and its overexpression was known to enhance the invasion ability in our previous study. This work demonstrated that the H-NS protein, a pleiotropic regulator of gene expression, was a new transcriptional modulator of evpP gene. The results showed that in vivo the transcriptional level of evpP was downregulated by H-NS and in vitro this global regulator interacted directly with evpP promoter region. Moreover, DNase I footprinting experiments mapping the interaction regions of H-NS and evpP revealed that this global regulator bound to evpP promoter and neighbouring areas at multiple sites. We provided a new insight into evpP regulation network and demonstrated the repression of H-NS to the transcription of evpP gene.
Recently, the devastating fish disease edwardsiellosis caused by Edwardsiella tarda has been widely concerned. The xenogeneic silencing of the classic regulator H-NS to the T6SS secretory protein EvpP, which played an important role in the virulence of Edw. tarda, was firstly reported in this study. It raised a better understanding of the virulence regulation of EvpP and provided more information about the complex infection mechanism of this pathogen. Our findings would contribute to the development of live attenuated vaccines against edwardsiellosis thus reducing the economic losses caused by this bacterium.
鱼类病原菌迟缓爱德华氏菌中的evpP基因编码T6SS分泌蛋白EvpP,并携带一个evpA-evpO独立启动子区域,对宿主细胞侵袭至关重要。evpP的转录受双组分系统EsrA-EsrB或铁浓度的正向调控,在我们之前的研究中已知其过表达可增强侵袭能力。这项工作表明,作为基因表达多效调节因子的H-NS蛋白是evpP基因的一种新的转录调节因子。结果表明,在体内H-NS下调evpP的转录水平,在体外这种全局调节因子直接与evpP启动子区域相互作用。此外,DNase I足迹实验绘制了H-NS与evpP的相互作用区域,结果显示这种全局调节因子在多个位点与evpP启动子及邻近区域结合。我们对evpP调控网络有了新的认识,并证明了H-NS对evpP基因转录的抑制作用。
最近,由迟缓爱德华氏菌引起的毁灭性鱼类疾病爱德华氏菌病受到广泛关注。本研究首次报道了经典调节因子H-NS对T6SS分泌蛋白EvpP的异源沉默,EvpP在迟缓爱德华氏菌的毒力中起重要作用。这加深了对EvpP毒力调控的理解,并提供了有关该病原菌复杂感染机制的更多信息。我们的发现将有助于开发针对爱德华氏菌病的减毒活疫苗,从而减少这种细菌造成的经济损失。