Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
Michiel Van Gent
Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
Joris P. Jansen
Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
Lisette M. Scheepmaker
Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
Anneroos Velthuizen
Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
Carla J.C. De Haas
Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
Kok P.M. Van Kessel
Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
Bart W. Bardoel
Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
Michael Boettcher
Department of Microbiology and Immunology, UCSF Diabetes Center, Keck Center for Noncoding RNA, University of California, San Francisco, San Francisco, CA 94143, USA
Michael T. McManus
Department of Microbiology and Immunology, UCSF Diabetes Center, Keck Center for Noncoding RNA, University of California, San Francisco, San Francisco, CA 94143, USA
Jos A.G. Van Strijp
Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
Robert Jan Lebbink
Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
Pieter-Jan A. Haas
Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
András N. Spaan
Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
Staphylococcal bi-component pore-forming toxins, also known as leukocidins, target and lyse human phagocytes in a receptor-dependent manner. S-components of the leukocidins Panton-Valentine leukocidin (PVL), γ-haemolysin AB (HlgAB) and CB (HlgCB), and leukocidin ED (LukED) specifically employ receptors that belong to the class of G-protein coupled receptors (GPCRs). Although these receptors share a common structural architecture, little is known about the conserved characteristics of the interaction between leukocidins and GPCRs. In this study, we investigated host cellular pathways contributing to susceptibility towards S. aureus leukocidin cytotoxicity. We performed a genome-wide CRISPR/Cas9 library screen for toxin-resistance in U937 cells sensitized to leukocidins by ectopic expression of different GPCRs. Our screen identifies post-translational modification (PTM) pathways involved in the sulfation and sialylation of the leukocidin-receptors. Subsequent validation experiments show differences in the impact of PTM moieties on leukocidin toxicity, highlighting an additional layer of refinement and divergence in the staphylococcal host-pathogen interface. Leukocidin receptors may serve as targets for anti-staphylococcal interventions and understanding toxin-receptor interactions will facilitate the development of innovative therapeutics. Variations in the genes encoding PTM pathways could provide insight into observed differences in susceptibility of humans to infections with S. aureus.
