Genome-wide CRISPR screen reveals novel host factors required for Staphylococcus aureus ?-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 (?HL) 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 ?HL susceptibility in human myeloid cells. We found gRNAs for ten genes enriched after intoxication with ?HL and focused on the top five hits. Besides a disintegrin and metalloproteinase domain-containing protein 10 (ADAM10), the host receptor for ?HL, 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 ?HL intoxication, but have only a slightly reduced ADAM10 surface expression. SGMS1 regulates lipid raft formation, suggesting that ?HL requires these membrane microdomains for attachment and cytotoxicity.

SUBMITTER: Virreira Winter S 

PROVIDER: S-EPMC4828653 | biostudies-literature | 2016 Apr

REPOSITORIES: biostudies-literature