The United Graduate School of Veterinary Science, Yamaguchi University, Yamaguchi, Japan.
Joint Faculty of Veterinary Medicine, Laboratory of Veterinary Public Health, Yamaguchi University, Yamaguchi, Japan.
Microbiologyopen. 2019 Apr;8(4):e00684. doi: 10.1002/mbo3.684. Epub 2018 Jul 5.
Francisella tularensis is the causative agent of the infectious disease tularemia and is designated a category A bioterrorism agent. The type VI secretion system encoded by the Francisella pathogenicity island (FPI) is necessary for intracellular growth; however, the functions of FPI proteins are largely unknown. In this study, we found that the FPI protein intracellular growth locus E (IglE) showed a unique localization pattern compared to other FPI proteins. Deleting iglE from Francisella tularensis subsp. novicida (F. novicida) decreased intracellular growth. Immunoprecipitation and pull-down assays revealed that IglE was associated with β-tubulin. Additionally, GFP-fused IglE colocalized with microtubule organizing centers (MTOCs) in 293T cells. The iglE deletion mutant was transferred with dynein toward MTOCs and packed into lysosome-localizing areas. Conversely, the wild-type F. novicida exhibited intracellular growth distant from MTOCs. In addition, IglE expressed in 293T cells colocalized with dynein. These results suggest that IglE helps to prevent dynein- and MTOC-mediated intracellular trafficking in host cells to inhibit the transport of F. novicida toward lysosomes.
土拉弗朗西斯菌是引起传染性疾病土拉菌病的病原体,被指定为 A 类生物恐怖主义剂。弗朗西斯氏菌致病性岛 (FPI) 编码的 VI 型分泌系统对于细胞内生长是必要的;然而,FPI 蛋白的功能在很大程度上是未知的。在这项研究中,我们发现与其他 FPI 蛋白相比,FPI 蛋白细胞内生长定位 E (IglE) 表现出独特的定位模式。从弗朗西斯氏菌 novicida 亚种(F. novicida)中删除 iglE 会降低细胞内生长。免疫沉淀和下拉测定显示 IglE 与 β-微管蛋白相关。此外,GFP 融合的 IglE 在 293T 细胞中与微管组织中心 (MTOC) 共定位。iglE 缺失突变体被向 MTOC 转移并被包装到溶酶体定位区域。相反,野生型 F. novicida 表现出远离 MTOC 的细胞内生长。此外,在 293T 细胞中表达的 IglE 与动力蛋白共定位。这些结果表明,IglE 有助于防止宿主细胞中动力蛋白和 MTOC 介导的细胞内运输,以抑制 F. novicida 向溶酶体的运输。