Tan Y P, Zheng J, Tung S L, Rosenshine I, Leung K Y
Department of Biological Sciences, Faculty of Science, National University of Singapore, Science Drive 4, Singapore 117543.
Department of Molecular Genetics and Biotechnology, Faculty of Medicine, The Hebrew University, PO Box 12272, Jerusalem 91120, Israel.
Microbiology (Reading). 2005 Jul;151(Pt 7):2301-2313. doi: 10.1099/mic.0.28005-0.
Edwardsiella tarda is a Gram-negative enteric bacterium affecting both animals and humans. Recently, a type III secretion system (TTSS) was found in Ed. tarda. Such systems are generally used by bacterial pathogens to deliver virulence factors into host cells to subvert normal cell functions. Genome-walking was performed from the eseB and esrB genes (homologues of Salmonella sseB and ssrB, respectively) identified in previous studies, to determine the sequences of the TTSS. Thirty-five ORFs were identified which encode the TTSS apparatus, chaperones, effectors and regulators. Mutants affected in genes representing each category were generated and found to have decreased survival and growth in fish phagocytes. LD(50) values of the mutants were increased by at least 10-fold in comparison to those of the wild-type strain. The adherence and invasion rates of the esrA and esrB mutants were enhanced while those of the other mutants remained similar to the wild-type. The eseC and eseD mutants showed slight autoaggregation in Dulbecco's Modified Eagle Medium, whereas the rest of the mutants failed to autoaggregate. Regulation of the TTSS was found to involve the two-component regulatory system esrA-esrB. This study showed that the TTSS is important for Ed. tarda pathogenesis. An understanding of this system will provide greater insight into the virulence mechanisms of this bacterial pathogen.
迟缓爱德华氏菌是一种革兰氏阴性肠道细菌,可感染动物和人类。最近,在迟缓爱德华氏菌中发现了III型分泌系统(TTSS)。这类系统通常被细菌病原体用来将毒力因子传递到宿主细胞中,以颠覆正常的细胞功能。从先前研究中鉴定出的eseB和esrB基因(分别是沙门氏菌sseB和ssrB的同源物)开始进行基因组步移,以确定TTSS的序列。共鉴定出35个开放阅读框,它们编码TTSS装置、分子伴侣、效应蛋白和调节蛋白。构建了代表每个类别的基因发生突变的突变体,发现它们在鱼类吞噬细胞中的存活和生长能力下降。与野生型菌株相比,突变体的半数致死剂量(LD50)值增加了至少10倍。esrA和esrB突变体的黏附率和侵袭率增强,而其他突变体的黏附率和侵袭率与野生型相似。eseC和eseD突变体在杜氏改良伊格尔培养基中表现出轻微的自聚集现象,而其他突变体则没有自聚集现象。发现TTSS的调节涉及双组分调节系统esrA-esrB。这项研究表明,TTSS对迟缓爱德华氏菌的致病作用很重要。对该系统的了解将为深入了解这种细菌病原体的毒力机制提供更多信息。