Wassenaar T M, Bleumink-Pluym N M, Newell D G, Nuijten P J, van der Zeijst B A
Department of Bacteriology, School of Veterinary Medicine, University of Utrecht, The Netherlands.
Infect Immun. 1994 Sep;62(9):3901-6. doi: 10.1128/iai.62.9.3901-3906.1994.
Campylobacter jejuni 81116 has two genes coding for flagellin, flaA and flaB. Fully motile wild-type C. jejuni bacteria express the flaA gene, with no flaB message being detected. A nonmotile flaA flaB+ mutant, R1, produced detectable levels of flagellin B which was incorporated into truncated flagella. After R1 had invaded INT-407 cells, a variant with increased motility, R1-V2, was isolated. R1-V2 produced full-length flagella and an increased amount of flagellin B. Transcriptional analysis showed that R1-V2 contained more flaB mRNA than its parental strain, R1. The flaB gene promoter sequence and primer extension experiments confirmed that transcription of the flaB gene is initiated from a sigma 54 promoter. Neither the promoter sequence nor the coding sequence of flaB had changed in R1-V2. In contrast to R1, R1-V2 no longer produced (truncated) flaA mRNA. The sigma 28 flaA promoter sequence was not changed in R1-V2. We propose that expression of the two flagellin genes in C. jejuni 81116 is regulated at the transcriptional level, in such a way that predominantly one gene at a time is transcribed. We compared the levels of invasiveness of the wild-type strain, R1, and R1-V2 for INT-407 cells. The shift in expression from flaA to flaB occurred not only during invasion assays but also under different conditions in the absence of eukaryotic cells.
空肠弯曲菌81116有两个编码鞭毛蛋白的基因,flaA和flaB。完全有运动能力的野生型空肠弯曲菌细菌表达flaA基因,未检测到flaB信息。一种无运动能力的flaA flaB+突变体R1产生了可检测水平的鞭毛蛋白B,该蛋白被整合到截短的鞭毛中。R1侵入INT-407细胞后,分离出一种运动能力增强的变体R1-V2。R1-V2产生全长鞭毛和增加量的鞭毛蛋白B。转录分析表明,R1-V2比其亲本菌株R1含有更多的flaB mRNA。flaB基因启动子序列和引物延伸实验证实,flaB基因的转录起始于一个σ54启动子。R1-V2中flaB的启动子序列和编码序列均未改变。与R1不同,R1-V2不再产生(截短的)flaA mRNA。R1-V2中σ28 flaA启动子序列未改变。我们提出,空肠弯曲菌81116中两个鞭毛蛋白基因的表达在转录水平上受到调控,使得一次主要转录一个基因。我们比较了野生型菌株、R1和R1-V2对INT-407细胞的侵袭水平。从flaA到flaB的表达转变不仅发生在侵袭试验期间,也发生在没有真核细胞的不同条件下。