Systematic Identification of Host Cell Regulators of Legionella pneumophila Pathogenesis Using a Genome-wide CRISPR Screen
| Autor: | Edwin E. Jeng, Haley Gause, Michael C. Bassik, Ruiqi Jian, Lihua Jiang, Nevan J. Krogan, Erica Stevenson, Danielle L. Swaney, Michael Snyder, Nnejiuwa U Ibe, Shaeri Mukherjee, David Jimenez-Morales, Varun Bhadkamkar, Joanne Chan |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Virulence Factors
Host–pathogen interaction Endoplasmic Reticulum Microbiology Legionella pneumophila Article Cell Line 03 medical and health sciences Mice 0302 clinical medicine Bacterial Proteins Phagocytosis Virology CRISPR Animals Guanine Nucleotide Exchange Factors Humans Clustered Regularly Interspaced Short Palindromic Repeats Gene 030304 developmental biology 0303 health sciences biology Effector Intracellular parasite Endoplasmic reticulum Macrophages Membrane Proteins Proteins U937 Cells biology.organism_classification bacterial infections and mycoses Cell biology respiratory tract diseases HEK293 Cells RAW 264.7 Cells rab GTP-Binding Proteins Chaperone (protein) Vacuoles biology.protein bacteria Parasitology Legionnaires' Disease 030217 neurology & neurosurgery HeLa Cells |
| Zdroj: | Cell Host Microbe |
| ISSN: | 1934-6069 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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