Ohtani Kaori, Hayashi Hideo, Shimizu Tohru
Department of Infection Biology, Institute of Basic Medical Sciences, University of Tsukuba, 1-1-1 Tenohdai, Tsukuba, Ibaraki 305-8575, Japan.
Mol Microbiol. 2002 Apr;44(1):171-9. doi: 10.1046/j.1365-2958.2002.02863.x.
A Gram-positive anaerobic pathogen, Clostridium perfringens, causes clostridial myonecrosis or gas gangrene in humans by producing numerous extracellular toxins and enzymes that act in concert to degrade host tissues. C. perfringens possesses a homologue of the luxS gene that is reported to be responsible for the production of autoinducer 2 (AI-2), which participates in quorum sensing in bacteria. The luxS mutant was constructed using C. perfringens strain 13, and the role of the luxS gene in toxin production was examined. The cell-free culture supernatant from wild-type strain 13 greatly stimulated the luminescence of Vibrio harveyi BB170, whereas that from the luxS mutant caused no significant stimulation, indicating that the luxS gene is necessary for AI-2 production in C. perfringens. The luxS mutant showed a reduced level of production of alpha-, kappa- and theta-toxins. In the luxS mutant, the transcription of the theta-toxin gene (pfoA) was lower at mid-exponential growth phase, whereas alpha- and kappa-toxin gene transcription was not significantly affected. The production of toxins in the luxS mutant was stimulated by the addition of the culture supernatant from the wild-type cells, possibly because of the presence of AI-2. Moreover, the expression of the pfoA gene in the luxS mutant was apparently activated when the mutant cells were cultured in the presence of culture supernatants from the wild-type C. perfringens, Escherichia coli DH5alpha carrying the luxS gene of C. perfringens. A deletion analysis of the luxS operon showed that the luxS gene alone is responsible for cell-cell signalling, and that the metB or cysK genes located upstream of luxS are not involved in regulating toxin production. Our results indicate that cell-cell signalling by AI-2 plays an important role in the regulation of toxin production in C. perfringens.
革兰氏阳性厌氧病原体产气荚膜梭菌通过产生多种细胞外毒素和酶协同作用降解宿主组织,从而导致人类发生梭菌性肌坏死或气性坏疽。产气荚膜梭菌拥有luxS基因的同源物,据报道该基因负责自诱导物2(AI-2)的产生,AI-2参与细菌的群体感应。使用产气荚膜梭菌13型菌株构建了luxS突变体,并研究了luxS基因在毒素产生中的作用。野生型13型菌株的无细胞培养上清液极大地刺激了哈维氏弧菌BB170的发光,而luxS突变体的无细胞培养上清液则未引起明显刺激,这表明luxS基因是产气荚膜梭菌产生AI-2所必需的。luxS突变体的α-、κ-和θ-毒素产生水平降低。在luxS突变体中,θ-毒素基因(pfoA)在指数生长中期的转录较低,而α-和κ-毒素基因的转录未受到明显影响。添加野生型细胞的培养上清液可刺激luxS突变体中毒素的产生,这可能是由于存在AI-2。此外,当luxS突变体细胞在野生型产气荚膜梭菌、携带产气荚膜梭菌luxS基因的大肠杆菌DH5α的培养上清液存在下培养时,pfoA基因的表达明显被激活。对luxS操纵子的缺失分析表明,仅luxS基因负责细胞间信号传导,位于luxS上游的metB或cysK基因不参与调节毒素产生。我们的结果表明,AI-2介导的细胞间信号传导在产气荚膜梭菌毒素产生的调节中起重要作用。