Infectious Diseases and Immunology Division, Indian Institute of Chemical Biology, Council of Scientific and Industrial Research, Kolkata, India.
J Bacteriol. 2013 May;195(9):2004-10. doi: 10.1128/JB.02127-12. Epub 2013 Feb 22.
Vibrio cholerae, a noninvasive bacterium, colonizes the intestinal epithelium and secretes cholera toxin (CT), a potent enterotoxin that causes the severe fluid loss characteristic of the disease cholera. In this study, we demonstrate that adherence of V. cholerae to the intestinal epithelial cell line INT 407 strongly induces the expression of the major virulence genes ctxAB and tcpA and the virulence regulatory gene toxT. No induction of toxR and tcpP, which encode transcriptional activators of toxT, was observed in adhered bacteria, and the adherence-dependent upregulation of toxT expression was independent of ToxR and TcpP. A sharp increase in the expression of the vieA gene, which encodes a cyclic di-GMP (c-di-GMP) phosphodiesterase, was observed in INT 407-adhered V. cholerae immediately after infection. Induction of toxT, ctxAB, and tcpA in INT 407-adhered vieA mutant strain O395 ΔvieA was consistently lower than in the parent strain, although no effect was observed in unadhered bacteria, suggesting that VieA has a role in the upregulation of toxT expression specifically in host cell-adhered V. cholerae. Furthermore, though VieA has both a DNA binding helix-turn-helix domain and an EAL domain conferring c-di-GMP phosphodiesterase activity, the c-di-GMP phosphodiesterase activity of VieA is necessary and sufficient for the upregulation of toxT expression.
霍乱弧菌是一种非侵入性细菌,定植于肠道上皮细胞并分泌霍乱毒素(CT),这种强效肠毒素导致霍乱这种疾病的严重液体流失。在这项研究中,我们证明了霍乱弧菌对肠道上皮细胞系 INT 407 的黏附强烈诱导了主要毒力基因 ctxAB 和 tcpA 以及毒力调节基因 toxT 的表达。在黏附细菌中没有观察到编码 toxT 转录激活物的 toxR 和 tcpP 的诱导,并且 toxT 表达的黏附依赖性上调独立于 ToxR 和 TcpP。在 INT 407 黏附的霍乱弧菌感染后立即观察到编码环二鸟苷酸(c-di-GMP)磷酸二酯酶的 vieA 基因的表达急剧增加。INT 407 黏附的 vieA 突变株 O395 ΔvieA 中 toxT、ctxAB 和 tcpA 的诱导始终低于亲本菌株,尽管在未黏附细菌中没有观察到这种现象,这表明 VieA 在宿主细胞黏附的霍乱弧菌中特异地上调 toxT 表达中起作用。此外,尽管 VieA 具有 DNA 结合螺旋-转角-螺旋结构域和赋予 c-di-GMP 磷酸二酯酶活性的 EAL 结构域,但 VieA 的 c-di-GMP 磷酸二酯酶活性对于 toxT 表达的上调是必需和充分的。