Systematic Identification of Host Cell Regulators of Legionella pneumophila Pathogenesis Using a Genome-wide CRISPR Screen
Systematic Identification of Host Cell Regulators of Legionella pneumophila Pathogenesis Using a Genome-wide CRISPR Screen
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DOI：
10.1016/j.chom.2019.08.017
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发表时间：
2019-10-09
影响因子：
30.3
通讯作者：
Bassik, Michael C.
中科院分区：
文献类型：
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作者：
Jeng, Edwin E.;Bhadkamkar, Varun;Bassik, Michael C.
在感染过程中，嗜肺军团菌将超过300种效应蛋白易位到宿主细胞质中，使病原体能够建立支持细菌复制的内质网（ER）样含军团菌空泡（LCV）。在这里，我们在U937人单核细胞/巨噬细胞样细胞中进行了全基因组CRISPR-Cas9筛选和二次靶向筛选，以系统地鉴定调节L.嗜肺菌屏幕显示已知的宿主因子被L. pneumophila，以及跨越不同的运输和信号转导途径的基因，以前没有连接到L。嗜肺菌发病机制我们进一步表征了C1 orf 43和KIAA 1109作为吞噬作用的调节剂，并表明RAB 10及其伴侣RABIF是最佳L.嗜肺菌复制和ER向LCV的募集。最后，我们发现Rab 10蛋白被招募到LCV并被效应子SidC/SdcA泛素化。总的来说，我们的研究结果提供了丰富的以前未描述的见解L。嗜肺菌发病机制和哺乳动物细胞功能。
During infection, Legionella pneumophila translocates over 300 effector proteins into the host cytosol, allowing the pathogen to establish an endoplasmic reticulum (ER)-like Legionella-containing vacuole (LCV) that supports bacterial replication. Here, we perform a genome-wide CRISPR-Cas9 screen and secondary targeted screens in U937 human monocyte/macrophage-like cells to systematically identify host factors that regulate killing by L. pneumophila. The screens reveal known host factors hijacked by L. pneumophila, as well as genes spanning diverse trafficking and signaling pathways previously not linked to L. pneumophila pathogenesis. We further characterize C1orf43 and KIAA1109 as regulators of phagocytosis and show that RAB10 and its chaperone RABIF are required for optimal L. pneumophila replication and ER recruitment to the LCV. Finally, we show that Rab10 protein is recruited to the LCV and ubiquitinated by the effectors SidC/SdcA. Collectively, our results provide a wealth of previously undescribed insights into L. pneumophila pathogenesis and mammalian cell function.