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Applying an Inducible Expression System to Study Interference of Bacterial Virulence Factors with Intracellular Signaling
Published on: June 25, 2015
The pathogenic E. coli type III effector EspZ interacts with host CD98 and facilitates host cell prosurvival
Stephanie R Shames1, Wanyin Deng, Julian A Guttman
1Michael Smith Laboratories, University of British Columbia, Vancouver, British Columbia, Canada.
Enteropathogenic Escherichia coli (EPEC) uses the effector EspZ to protect host cells from damage. EspZ interacts with host CD98, promoting cell survival mechanisms involving focal adhesion kinase (FAK).
Area of Science:
- Microbiology
- Cell Biology
- Pathogen-Host Interactions
Background:
- Enterohaemorrhagic and enteropathogenic Escherichia coli (EHEC/EPEC) are diarrheal pathogens causing attaching and effacing (A/E) lesions.
- These bacteria utilize a type III secretion system (T3SS) to inject virulence effector proteins into host cells.
Purpose of the Study:
- To investigate the function of the T3SS effector EspZ in EPEC infection.
- To elucidate the interaction between EspZ and host cell proteins and its role in virulence.
Main Methods:
- Yeast two-hybrid assays
- Proteomics
- Immunofluorescence microscopy
- Co-immunoprecipitation
- Cell cytotoxicity assays (HeLa, MDCK cells)
Main Results:
- EPEC lacking EspZ (EPEC DeltaespZ) exhibited increased cytotoxicity compared to wild-type EPEC.
- EspZ interacts with the host protein CD98, which confers protection against EPEC-induced cytotoxicity.
- EspZ enhances phosphorylation of focal adhesion kinase (FAK) and AKT; CD98 facilitates FAK phosphorylation.
Conclusions:
- EspZ is a key EPEC effector that promotes host cell survival.
- The EspZ-CD98 interaction is crucial for mitigating pathogen-induced cytotoxicity.
- Host cell survival mechanisms involving FAK are promoted by EspZ and CD98 during A/E pathogen infection.
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