Complete and ubiquitinated proteome of the Legionella-containing vacuole within human macrophages.

Complete and ubiquitinated proteome of the Legionella-containing vacuole within human macrophages.
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DOI:
10.1021/pr500765x
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发表时间:
2015-01-02
影响因子:
4.4
通讯作者:
Abu Kwaik, Yousef
Abu Kwaik, Yousef
中科院分区:
生物学2区
文献类型:
--
作者:
Bruckert, William M.;Abu Kwaik, Yousef

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在原生动物或人类巨噬细胞内,嗜肺军团菌逃避内体途径,并在称为含军团菌液泡 (LCV) 的内质网衍生液泡内复制。嗜肺军团菌的 LCV 膜定位 AnkB 效应子是一种 F-box 蛋白,可介导 LCV 与赖氨酸 48 连接的多聚泛素化蛋白的修饰,这对于液泡内复制至关重要。使用高通量 LC-MS 分析,我们鉴定了从人 U937 巨噬细胞中纯化的 LCV 的总蛋白质组和泛素化宿主来源蛋白质组。携带AA100/130b WT菌株的LCV含有1193个蛋白质,其中24个泛素化蛋白质,而ankB突变体LCV含有1546个蛋白质,其中29个泛素化蛋白质。通路分析揭示了 WT 和 ankB 突变型 LCV 上参与信号传导、蛋白质转运、磷脂酰肌醇和碳水化合物代谢的蛋白质的富集。 ankB 突变体 LCV 优先富集参与转录/翻译和免疫反应的蛋白质。 WT 株 LCV 上的泛素化蛋白富集于免疫反应、信号传导、调节、细胞内运输和氨基酸运输途径,而 ankB 突变体 LCV 上的泛素化蛋白则富集于囊泡运输、信号传导和泛素化途径。人类巨噬细胞内完整且泛素化的 LCV 蛋白质组阐明了 LCV 复杂且动态的生物发生,并为未来的研究提供了丰富的资源。
Within protozoa or human macrophages Legionella pneumophila evades the endosomal pathway and replicates within an ER-derived vacuole termed the Legionella-containing vacuole (LCV). The LCV membrane-localized AnkB effector of L. pneumophila is an F-box protein that mediates decoration of the LCV with lysine48-linked polyubiquitinated proteins, which is essential for intravacuolar replication. Using high-throughput LC–MS analysis, we have identified the total and ubiquitinated host-derived proteome of LCVs purified from human U937 macrophages. The LCVs harboring the AA100/130b WT strain contain 1193 proteins including 24 ubiquitinated proteins, while the ankB mutant LCVs contain 1546 proteins with 29 ubiquitinated proteins. Pathway analyses reveal the enrichment of proteins involved in signaling, protein transport, phosphatidylinositol, and carbohydrate metabolism on both WT and ankB mutant LCVs. The ankB mutant LCVs are preferentially enriched for proteins involved in transcription/translation and immune responses. Ubiquitinated proteins on the WT strain LCVs are enriched for immune response, signaling, regulation, intracellular trafficking, and amino acid transport pathways, while ubiquitinated proteins on the ankB mutant LCVs are enriched for vesicle trafficking, signaling, and ubiquitination pathways. The complete and ubiquitinated LCV proteome within human macrophages illustrates complex and dynamic biogenesis of the LCV and provides a rich resource for future studies.
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