Kanamaru K, Kanamaru K, Tatsuno I, Tobe T, Sasakawa C
Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-0071, Japan.
Mol Microbiol. 2000 Nov;38(4):805-16. doi: 10.1046/j.1365-2958.2000.02171.x.
The quorum-sensing system in bacteria is a well-known regulatory system that controls gene expression in a cell density-dependent manner. A transcriptional regulator (LuxR homologue), signal synthase (LuxI homologue) and autoinducer (acyl homoserine lactone) are indispensable for this system in most Gram-negative bacteria. In this study, we found that SdiA, an Escherichia coli LuxR homologue, is a negative regulator of the expression of virulence factors EspD and intimin in enterohaemorrhagic E. coli (EHEC) O157:H7. The expression of EspD and intimin was inhibited at the RNA level upon SdiA overexpression. SdiA has a DNA-binding motif in its C-terminal part and can bind to the promoter regions of the esp and eae genes in vitro. Extracellular factors, which accumulate in culture supernatants of O157:H7 at the stationary phase of growth and inhibit EspD and intimin synthesis, bind to the N-terminal part of SdiA in vivo and in vitro. O157:H7 overproducing the N-terminal part of SdiA exhibited hypertranscription of EspD and intimin, suggesting that the overproduced N-terminal part had inhibited the activity of intact SdiA through titration of the extracellular factors. These results indicate that a quorum-sensing system including the SdiA protein controls colonization by O157:H7.
细菌中的群体感应系统是一种众所周知的调节系统,它以细胞密度依赖的方式控制基因表达。转录调节因子(LuxR同源物)、信号合成酶(LuxI同源物)和自诱导物(酰基高丝氨酸内酯)对于大多数革兰氏阴性细菌中的该系统而言不可或缺。在本研究中,我们发现大肠杆菌LuxR同源物SdiA是肠出血性大肠杆菌(EHEC)O157:H7中毒力因子EspD和紧密黏附素表达的负调节因子。SdiA过表达时,EspD和紧密黏附素的表达在RNA水平受到抑制。SdiA在其C末端部分具有一个DNA结合基序,并且在体外能够与esp和eae基因的启动子区域结合。在生长稳定期积聚于O157:H7培养上清液中并抑制EspD和紧密黏附素合成的细胞外因子,在体内和体外均能与SdiA的N末端部分结合。过量产生SdiA N末端部分的O157:H7表现出EspD和紧密黏附素的超转录,这表明过量产生的N末端部分通过滴定细胞外因子抑制了完整SdiA的活性。这些结果表明,包括SdiA蛋白在内的群体感应系统控制着O157:H7的定植。