Global mapping of Salmonella enterica-host protein-protein interactions during infection
Description
Summary of the study: Intracellular 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 32 Salmonella enterica serovar Typhimurium (STm) strains expressing chromosomally encoded affinity-tagged effectors, 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 used this PPI resource to further demonstrate that SseJ, SseL and SifA modulate cholesterol accumulation at the Salmonella Containing Vacuole (SCV) partially via the cholesterol transporter Niemann-Pick C1 protein (NPC1), PipB recruits the organelle contact site protein PDZD8 to the SCV, and SteC promotes actin bundling by phosphorylating formin-like proteins. Annotation to this repository: This collection of supplementary and original data completes the associated manuscript of the same title. It comprises: (01-04) The enrichment plots of the large scale AP/QMS study (related to figures 2, 3 and S2) (05-06) The abundances of all proteins in the AP/QMS study, alongside the full limma results (Figures 2, 3, S2 and S4) (07) The data at the basis of all plots shown in the manuscript (Figures 4, 5, 6, 7, S6 and S7) (08) All original Western Blot scans (Figures 4, 6, 7, S1, S3, S5, S7 and S8) (09) The limma results of the validation in pBMDMs (Figure S5) (10) The limma results of the separate TMT-run to assess the interaction partners of SseJ (Figures 5 and S6) (11-12) Additional information about the antibodies and primers used in this study (KRT and Experimental Procedures)