Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-Universität, München, Germany.
PLoS One. 2012;7(4):e35341. doi: 10.1371/journal.pone.0035341. Epub 2012 Apr 6.
Helicobacter pylori, the causative agent of type B gastritis, peptic ulcers, gastric adenocarcinoma and MALT lymphoma, uses the Cag type IV secretion system to induce a strong proinflammatory response in the gastric mucosa and to inject its effector protein CagA into gastric cells. CagA translocation results in altered host cell gene expression profiles and cytoskeletal rearrangements, and it is considered as a major bacterial virulence trait. Recently, it has been shown that binding of the type IV secretion apparatus to integrin receptors on target cells is a crucial step in the translocation process. Several bacterial proteins, including the Cag-specific components CagL and CagI, have been involved in this interaction. Here, we have examined the localization and interactions of CagI in the bacterial cell. Since the cagI gene overlaps and is co-transcribed with the cagL gene, the role of CagI for type IV secretion system function has been difficult to assess, and conflicting results have been reported regarding its involvement in the proinflammatory response. Using a marker-free gene deletion approach and genetic complementation, we show now that CagI is an essential component of the Cag type IV secretion apparatus for both CagA translocation and interleukin-8 induction. CagI is distributed over soluble and membrane-associated pools and seems to be partly surface-exposed. Deletion of several genes encoding essential Cag components has an impact on protein levels of CagI and CagL, suggesting that both proteins require partial assembly of the secretion apparatus. Finally, we show by co-immunoprecipitation that CagI and CagL interact with each other. Taken together, our results indicate that CagI and CagL form a functional complex which is formed at a late stage of secretion apparatus assembly.
幽门螺杆菌是 B 型胃炎、消化性溃疡、胃腺癌和 MALT 淋巴瘤的病原体,它利用 Cag 型 IV 型分泌系统在胃黏膜中诱导强烈的促炎反应,并将其效应蛋白 CagA 注射到胃细胞中。CagA 的易位导致宿主细胞基因表达谱和细胞骨架重排的改变,被认为是细菌主要的毒力特征。最近,已经表明,IV 型分泌装置与靶细胞上整合素受体的结合是易位过程中的关键步骤。几种细菌蛋白,包括 Cag 特异性成分 CagL 和 CagI,已经参与了这种相互作用。在这里,我们研究了 CagI 在细菌细胞中的定位和相互作用。由于 cagI 基因与 cagL 基因重叠并共转录,因此 CagI 对 IV 型分泌系统功能的作用很难评估,并且关于其参与促炎反应的结果存在冲突。使用无标记基因缺失方法和遗传互补,我们现在表明 CagI 是 Cag 型 IV 型分泌装置的一个重要组成部分,对于 CagA 的易位和白细胞介素-8 的诱导都是必需的。CagI 分布在可溶性和膜相关的池中和部分表面暴露。缺失几个编码必需 Cag 成分的基因会影响 CagI 和 CagL 的蛋白水平,这表明这两种蛋白都需要分泌装置的部分组装。最后,我们通过共免疫沉淀表明 CagI 和 CagL 相互作用。总之,我们的结果表明 CagI 和 CagL 形成了一个功能复合物,该复合物在分泌装置组装的后期形成。