1. Global mapping ofSalmonella entericahost protein-protein interactions during infection
- Author
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Joel Selkrig, Klemens Rottner, Olivia Steele-Mortimer, Cristina Viéitez, Leigh A. Knodler, Matthias Geyer, David W. Holden, Karoline Scholzen, Mikhail M. Savitski, Clement M. Potel, Keith C. Fernandez, Philipp Walch, Mandy Rettel, Frank Stein, Athanasios Typas, and HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.
- Subjects
Male ,Salmonella typhimurium ,Cell type ,PDZD8 ,protein-protein interactions ,Vacuole ,Interactome ,Microbiology ,Article ,Protein–protein interaction ,Mice ,Bacterial Proteins ,bacterial pathogen ,Virology ,actin bundling ,Organelle ,Animals ,Humans ,Protein Interaction Domains and Motifs ,Adaptor Proteins, Signal Transducing ,Bacteria ,biology ,Effector ,Macrophages ,Salmonella enterica ,Epithelial Cells ,biology.organism_classification ,Cell biology ,NPC1 ,RAW 264.7 Cells ,cholesterol trafficking ,Host-Pathogen Interactions ,Phosphorylation ,Female ,Parasitology ,FMNL ,Intracellular ,HeLa Cells ,effectors - Abstract
SummaryIntracellular bacterial pathogens inject effector proteins into host cells to hijack diverse cellular processes and promote their survival and proliferation. To systematically map effector-host protein-protein interactions (PPIs) during infection, we generated a library of 32Salmonella entericaserovar Typhimurium (STm) strains expressing chromosomally encoded affinity-tagged effector proteins, and quantified PPIs in macrophages and epithelial cells by Affinity-Purification Quantitative Mass-Spectrometry. Thereby, we identified 25 previously described and 421 novel effector-host PPIs. While effectors converged on the same host cellular processes, most had multiple targets, which often differed between cell types. Using reciprocal co-immunoprecipitations, we validated 13 out of 22 new PPIs. We then used this host-pathogen physical interactome resource to demonstrate that SseJ and SseL collaborate in redirecting cholesterol to theSalmonellaContaining Vacuole (SCV) via NPC1, PipB directly recruits the organelle contact site protein PDZD8 to the SCV, and SteC promotes actin bundling by directly phosphorylating formin-like proteins.
- Published
- 2021