title: Genome-wide CRISPR screen reveals novel host factors required for Staphylococcus aureus α-hemolysin-mediated toxicity.
abstract: Staphylococcus aureus causes a wide variety of infections and antibiotic resistant strains are a major problem in hospitals. One of the best studied virulence factors of S. aureus is the pore-forming toxin alpha hemolysin (αHL) whose mechanism of action is incompletely understood. We performed a genome-wide loss-of-function screen using CRISPR/Cas9 technology to identify host targets required for αHL susceptibility in human myeloid cells. We found gRNAs for ten genes enriched after intoxication with αHL and focused on the top five hits. Besides a disintegrin and metalloproteinase domain-containing protein 10 (ADAM10), the host receptor for αHL, we identified three proteins, Sys1 golgi trafficking protein (SYS1), ADP-ribosylation factor 1 (ARFRP1), and tetraspanin-14 (TSPAN14) which regulate the presentation of ADAM10 on the plasma membrane post-translationally. Interestingly, we also showed that cells lacking sphingomyelin synthase 1 (SGMS1) resist αHL intoxication, but have only a slightly reduced ADAM10 surface expression. SGMS1 regulates lipid raft formation, suggesting that αHL requires these membrane microdomains for attachment and cytotoxicity.
authors.affiliation: Department of Cellular Microbiology Max Planck Institute for Infection Biology, Charitéplatz 1, 10117 Berlin, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Virreira Winter
authors.affiliation: Department of Cellular Microbiology Max Planck Institute for Infection Biology, Charitéplatz 1, 10117 Berlin, Germany.
authors.forename: Arturo
authors.initials: A
authors.lastname: Zychlinsky
authors.affiliation: Department of Cellular Microbiology Max Planck Institute for Infection Biology, Charitéplatz 1, 10117 Berlin, Germany.
authors.forename: Bart W
authors.initials: BW
authors.lastname: Bardoel
mesh_terms: D001427:Bacterial Toxins / Q000633:toxicity*; D002460:Cell Line; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D005820:Genetic Testing; D006460:Hemolysin Proteins / Q000633:toxicity*; D054884:Host-Pathogen Interactions; D006801:Humans; D022423:Myeloid Cells / Q000187:drug effects* / Q000502:physiology; D011485:Protein Binding; D025941:Protein Interaction Mapping; D013211:Staphylococcus aureus / Q000472:pathogenicity*; D037521:Virulence Factors / Q000633:toxicity
chemical_list: D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D037521:Virulence Factors; C021091:staphylococcal alpha-toxin