Time-Resolved Fluorescence Microscopy Screens on Host Protein Subversion During Bacterial Cell Invasion

Area of Science:

• Microbiology
• Cell Biology
• Infectious Diseases

Background:

• Intracellular bacterial pathogens invade host cells by manipulating host signaling pathways, particularly vesicular trafficking.
• Host endomembrane compartments are dynamically reorganized during bacterial invasion and intracellular niche establishment.
• Shigella flexneri, a cause of bacillary dysentery, recruits host recycling and exocyst pathway components to facilitate entry into non-phagocytic enterocytes.

Purpose of the Study:

• To develop and detail protocols for time-resolved fluorescence microscopy screening to study host-pathogen interactions.
• To enable medium-to-high throughput multidimensional imaging of dynamic host-pathogen cross-talk.
• To provide a adaptable workflow for studying various host factor libraries and pathogen models using Shigella flexneri as a model.

Main Methods:

• Utilizing time-resolved screens by fluorescence microscopy to overcome challenges in monitoring asynchronous bacterial infection events.
• Employing libraries of fluorescently tagged host factors to identify key cellular components involved in host subversion.
• Developing detailed protocols for multidimensional imaging screening of host-pathogen interactions.

Main Results:

• Demonstrated the utility of fluorescence microscopy screening for studying host subversion by intracellular pathogens.
• Provided a robust workflow adaptable for different host factor libraries and pathogen models.
• Enabled dynamic monitoring of host-pathogen cross-talk during Shigella flexneri infection.

Conclusions:

• Time-resolved fluorescence microscopy is a powerful tool for dissecting host-pathogen interactions.
• The developed protocols facilitate the study of bacterial invasion mechanisms and intracellular pathogenesis.
• This approach can be broadly applied to understand host manipulation by various microbial pathogens.