BioID screen of Salmonella type 3 secreted effectors reveals host factors involved in vacuole positioning and stability during infection
| Autor: | Jonathan St-Germain, John H. Brumell, Estelle M.N. Laurent, Etienne Coyaud, Brian Raught, Kirsten C. Boddy, Vanessa M. D’Costa, Taoyingnan Li, Sergio Grinstein |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
Microbiology (medical)
Salmonella typhimurium Immunoprecipitation Immunology Dynein Virulence Biotin Vacuole Biology Applied Microbiology and Biotechnology Microbiology Protein–protein interaction 03 medical and health sciences Bacterial Proteins Salmonella Genetics Humans Cytoskeleton 030304 developmental biology 0303 health sciences Staining and Labeling 030306 microbiology Effector HEK 293 cells Intracellular Signaling Peptides and Proteins Cell Biology 16. Peace & justice Cell biology Protein Transport HEK293 Cells Gene Knockdown Techniques Host-Pathogen Interactions Salmonella Infections Vacuoles HeLa Cells |
| Zdroj: | Nature microbiology. 4(12) |
| ISSN: | 2058-5276 |
| Popis: | Many bacterial pathogens express virulence proteins that are translocated into host cells (herein referred to as effectors), where they can interact with target proteins to manipulate host cell processes. These effector-host protein interactions are often dynamic and transient in nature, making them difficult to identify using traditional interaction-based methods. Here, we performed a systematic comparison between proximity-dependent biotin labelling (BioID) and immunoprecipitation coupled with mass spectrometry to investigate a series of Salmonella type 3 secreted effectors that manipulate host intracellular trafficking (SifA, PipB2, SseF, SseG and SopD2). Using BioID, we identified 632 candidate interactions with 381 unique human proteins, collectively enriched for roles in vesicular trafficking, cytoskeleton components and transport activities. From the subset of proteins exclusively identified by BioID, we report that SifA interacts with BLOC-2, a protein complex that regulates dynein motor activity. We demonstrate that the BLOC-2 complex is necessary for SifA-mediated positioning of Salmonella-containing vacuoles, and affects stability of the vacuoles during infection. Our study provides insight into the coordinated activities of Salmonella type 3 secreted effectors and demonstrates the utility of BioID as a powerful, complementary tool to characterize effector-host protein interactions. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink