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弗朗西斯氏菌致病性岛蛋白细胞内生长区 E(IglE)在哺乳动物细胞中的表达涉及细胞内运输,可能通过微管组织中心。

Expression of Francisella pathogenicity island protein intracellular growth locus E (IglE) in mammalian cells is involved in intracellular trafficking, possibly through microtubule organizing center.

机构信息

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.

出版信息

Microbiologyopen. 2019 Apr;8(4):e00684. doi: 10.1002/mbo3.684. Epub 2018 Jul 5.

Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a30e/6460260/43afc55a7fac/MBO3-8-e00684-g001.jpg

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