2. Academic Validation
  3. 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

  • Cell Host Microbe. 2019 Oct 9;26(4):551-563.e6. doi: 10.1016/j.chom.2019.08.017.
Edwin E Jeng 1,  Varun Bhadkamkar 2,  Nnejiuwa U Ibe 2,  Haley Gause 2,  Lihua Jiang 3,  Joanne Chan 3,  Ruiqi Jian 3,  David Jimenez-Morales 4,  Erica Stevenson 4,  Nevan J Krogan 4,  Danielle L Swaney 4,  Michael P Snyder 3,  Shaeri Mukherjee 5,  Michael C Bassik 6
Affiliations

Affiliations

  • 1 Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA; Program in Cancer Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 2 Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94143, USA.
  • 3 Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 4 Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA, USA; Quantitative Biosciences Institute (QBI), University of California, San Francisco, San Francisco, CA, USA; The J. David Gladstone Institutes, San Francisco, CA, USA.
  • 5 Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94143, USA. Electronic address: [email protected].
  • 6 Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA. Electronic address: [email protected].
PMID: 31540829 DOI: 10.1016/j.chom.2019.08.017
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 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

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

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