Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.
Infect Immun. 2013 Apr;81(4):1334-40. doi: 10.1128/IAI.01377-12. Epub 2013 Feb 12.
The bacterial pathogen Listeria monocytogenes causes serious food-borne illnesses in pregnant women and the immunocompromised. L. monocytogenes promotes its internalization into host epithelial cells and then uses an F-actin-dependent motility process to spread from infected cells to surrounding healthy cells. In cultured enterocytes, efficient spread of L. monocytogenes requires the secreted bacterial protein InlC. InlC promotes dissemination by physically interacting with and antagonizing the function of the human adaptor protein Tuba. Here we examine the role of InlC and its interaction with host Tuba during infection in mice. The study took advantage of a single-amino-acid substitution (K173A) in InlC that impairs binding to human Tuba but does not affect InlC-mediated inhibition of the NF-κB pathway. Mice were inoculated intravenously with the wild-type L. monocytogenes strain EGD, an isogenic strain deleted for the inlC gene (ΔinlC), or a strain expressing K173A mutant InlC (inlC.K173A). The 50% lethal doses (LD(50)) for the ΔinlC or inlC.K173A mutant strain were approximately 4- or 6-fold greater than that for the wild-type strain, indicating a role for inlC in virulence. Compared to the wild-type strain, the inlC.K173A mutant strain exhibited lower bacterial loads in the liver. Histological analysis of livers indicated that the two inlC mutant strains produced smaller foci of infection than did the wild-type strain. These smaller foci are consistent with a role for InlC in cell-to-cell spread in vivo. Taken together, these results provide evidence that interaction of InlC with host Tuba is important for full virulence.
细菌病原体李斯特菌单核细胞增生李斯特菌会导致孕妇和免疫功能低下者严重的食源性疾病。李斯特菌单核细胞增生李斯特菌促进其内化到宿主上皮细胞中,然后利用依赖 F-肌动蛋白的运动过程从受感染的细胞传播到周围健康的细胞。在培养的肠细胞中,李斯特菌单核细胞增生李斯特菌的有效传播需要分泌的细菌蛋白 InlC。InlC 通过与人类衔接蛋白 Tuba 物理相互作用并拮抗其功能来促进传播。在这里,我们研究了 InlC 及其在感染小鼠过程中与宿主 Tuba 的相互作用。这项研究利用了 InlC 中的一个单氨基酸替换(K173A),该替换削弱了与人类 Tuba 的结合,但不影响 InlC 介导的 NF-κB 途径抑制。小鼠经静脉接种野生型李斯特菌 EGD 菌株、缺失 inlC 基因的同基因菌株(ΔinlC)或表达 K173A 突变体 InlC 的菌株(inlC.K173A)。ΔinlC 或 inlC.K173A 突变株的 50%致死剂量(LD 50)比野生型菌株约大 4 或 6 倍,表明 inlC 与毒力有关。与野生型菌株相比,inlC.K173A 突变株在肝脏中的细菌载量较低。肝脏组织学分析表明,与野生型菌株相比,两种 inlC 突变株产生的感染灶较小。这些较小的病灶与 InlC 在体内细胞间传播中的作用一致。综上所述,这些结果提供了证据表明 InlC 与宿主 Tuba 的相互作用对于充分的毒力很重要。