Cell Host & Microbe
Volume 26, Issue 4, 9 October 2019, Pages 551-563.e6
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Systematic Identification of Host Cell Regulators of Legionella pneumophila Pathogenesis Using a Genome-wide CRISPR Screen

https://doi.org/10.1016/j.chom.2019.08.017Get rights and content
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Highlights

  • CRISPR screen reveals host genes regulating distinct steps of L. pneumophila infection

  • Previously uncharacterized genes C1ORF43 and KIAA1109 regulate phagocytosis

  • Host Rab10 is hijacked by SidC/SdcA to promote ER recruitment and bacterial replication

Summary

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.

Keywords

Legionella pneumophila
CRISPR screen
RAB10
C1ORF43
phagocytosis
intracellular bacteria
pathogen
host-pathogen interaction

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Present address: Department of Medicine, Division of Cardiovascular Medicine, Stanford University, Stanford, CA 94305, USA

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