Xia Pengpeng, Song Yujie, Zou Yajie, Yang Ying, Zhu Guoqiang
College of Veterinary Medicine, Yangzhou University, Yangzhou, China.
Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, China.
J Basic Microbiol. 2015 Sep;55(9):1118-24. doi: 10.1002/jobm.201400901. Epub 2015 Apr 1.
The FaeG subunit is the major constituent of F4(+) fimbriae, associated with glycoprotein and/or glycolipid receptor recognition and majorly contributes to the pathogen attachment to the host cells. To investigate the key factor involved in the fimbrial binding of F4(+) Escherichia coli, both the recombinant E. coli SE5000 strains carrying the fae operon gene clusters that express the different types of fimbriae in vitro, named as rF4ab, rF4ac, and rF4ad, respectively, corresponding to the fimbrial types F4ab, F4ac, and F4ad, and the three isogenic in-frame faeG gene deletion mutants were constructed. The adhesion assays and adhesion inhibition assays showed that ΔfaeG mutants had a significant reduction in the binding to porcine brush border as well as the intestinal epithelial cell lines, while the complemented strain ΔfaeG/pfaeG restored the adhesion function. The recombinant bacterial strains rF4ab, rF4ac, and rF4ad have the same binding property as wild-type F4(+) E. coli strains do and improvement in terms of binding to porcine brush border and the intestinal epithelial cells, and the adherence was blocked by the monoclonal antibody anti-F4 fimbriae. These data demonstrate that the fimbrial binding of F4(+) E. coli is directly mediated by the major FaeG subunit.
FaeG亚基是F4(+)菌毛的主要组成部分,与糖蛋白和/或糖脂受体识别相关,主要促进病原体与宿主细胞的附着。为了研究参与F4(+)大肠杆菌菌毛结合的关键因素,构建了携带fae操纵子基因簇的重组大肠杆菌SE5000菌株,这些基因簇在体外表达不同类型的菌毛,分别命名为rF4ab、rF4ac和rF4ad,对应于菌毛类型F4ab、F4ac和F4ad,同时构建了三个同框faeG基因缺失突变体。黏附试验和黏附抑制试验表明,ΔfaeG突变体与猪刷状缘以及肠上皮细胞系的结合显著减少,而互补菌株ΔfaeG/pfaeG恢复了黏附功能。重组菌株rF4ab、rF4ac和rF4ad具有与野生型F4(+)大肠杆菌菌株相同的结合特性,并且在与猪刷状缘和肠上皮细胞的结合方面有所改善,并且抗F4菌毛单克隆抗体可阻断其黏附。这些数据表明,F4(+)大肠杆菌的菌毛结合直接由主要的FaeG亚基介导。