Institut Pasteur, Unité des interactions Bactéries-Cellules, Paris, France.
PLoS Pathog. 2011 Aug;7(8):e1002168. doi: 10.1371/journal.ppat.1002168. Epub 2011 Aug 4.
L. monocytogenes is a facultative intracellular bacterium responsible for listeriosis. It is able to invade, survive and replicate in phagocytic and non-phagocytic cells. The infectious process at the cellular level has been extensively studied and many virulence factors have been identified. Yet, the role of InlK, a member of the internalin family specific to L. monocytogenes, remains unknown. Here, we first show using deletion analysis and in vivo infection, that InlK is a bona fide virulence factor, poorly expressed in vitro and well expressed in vivo, and that it is anchored to the bacterial surface by sortase A. We then demonstrate by a yeast two hybrid screen using InlK as a bait, validated by pulldown experiments and immunofluorescence analysis that intracytosolic bacteria via an interaction with the protein InlK interact with the Major Vault Protein (MVP), the main component of cytoplasmic ribonucleoproteic particules named vaults. Although vaults have been implicated in several cellular processes, their role has remained elusive. Our analysis demonstrates that MVP recruitment disguises intracytosolic bacteria from autophagic recognition, leading to an increased survival rate of InlK over-expressing bacteria compared to InlK(-) bacteria. Together these results reveal that MVP is hijacked by L. monocytogenes in order to counteract the autophagy process, a finding that could have major implications in deciphering the cellular role of vault particles.
单核细胞增生李斯特菌是一种兼性细胞内细菌,负责李斯特菌病。它能够入侵、存活和复制吞噬细胞和非吞噬细胞。在细胞水平上的感染过程已经得到了广泛的研究,许多毒力因子已经被确定。然而,内部素家族成员 InlK 的作用,一种特定于单核细胞增生李斯特菌的内部素,仍然未知。在这里,我们首先通过缺失分析和体内感染显示,InlK 是一种真正的毒力因子,体外表达水平低,体内表达水平高,并且通过天冬酰胺内肽酶 A 将其锚定在细菌表面。然后,我们通过使用 InlK 作为诱饵的酵母双杂交筛选,通过下拉实验和免疫荧光分析进行验证,证明了通过与蛋白 InlK 的相互作用,胞质内细菌与主要穹窿蛋白(MVP)相互作用,MVP 是细胞质核糖核蛋白颗粒(称为穹窿)的主要成分。尽管穹窿已被涉及到几种细胞过程,但它们的作用仍然难以捉摸。我们的分析表明,MVP 的募集使胞质内细菌逃避自噬识别,导致过表达 InlK 的细菌比 InlK(-)细菌的存活率更高。这些结果表明,MVP 被单核细胞增生李斯特菌劫持,以对抗自噬过程,这一发现可能对破译穹窿颗粒的细胞作用具有重要意义。