1. Systematic Identification of Host Cell Regulators of Legionella pneumophila Pathogenesis Using a Genome-wide CRISPR Screen
- Author
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Jeng, Edwin E, Bhadkamkar, Varun, Ibe, Nnejiuwa U, Gause, Haley, Jiang, Lihua, Chan, Joanne, Jian, Ruiqi, Jimenez-Morales, David, Stevenson, Erica, Krogan, Nevan J, Swaney, Danielle L, Snyder, Michael P, Mukherjee, Shaeri, and Bassik, Michael C
- Subjects
Virulence Factors ,Immunology ,host-pathogen interaction ,Endoplasmic Reticulum ,Microbiology ,Cell Line ,Legionella pneumophila ,CRISPR screen ,Mice ,Bacterial Proteins ,Phagocytosis ,2.1 Biological and endogenous factors ,Animals ,Humans ,Guanine Nucleotide Exchange Factors ,Clustered Regularly Interspaced Short Palindromic Repeats ,Aetiology ,Lung ,intracellular bacteria ,Macrophages ,Proteins ,Membrane Proteins ,Pneumonia ,U937 Cells ,bacterial infections and mycoses ,respiratory tract diseases ,Infectious Diseases ,HEK293 Cells ,RAW 264.7 Cells ,rab GTP-Binding Proteins ,Medical Microbiology ,Hela Cells ,Vacuoles ,Pneumonia & Influenza ,bacteria ,RAB10 ,Legionnaires' Disease ,C1ORF43 ,Infection ,pathogen - Abstract
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 factorsthat 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 ofphagocytosis 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.
- Published
- 2019