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
