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Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking
Published on: December 17, 2013
Enteropathogenic Escherichia coli effector EspF interacts with host protein Abcf2.
Jean-Philippe Nougayrède1, Gregory H Foster, Michael S Donnenberg
1University of Maryland Baltimore, School of Medicine, Division of Infectious Diseases, 20 Penn Street, Baltimore, MD 21201, USA.
Cellular Microbiology
|October 27, 2006
Summary
Enteropathogenic Escherichia coli (EPEC) uses the EspF protein to disrupt host cells and trigger cell death. Researchers found that EspF interferes with Abcf2, a protein that normally protects cells, leading to increased apoptosis.
Area of Science:
- Microbiology
- Cell Biology
- Molecular Biology
Background:
- Enteropathogenic Escherichia coli (EPEC) causes infant diarrhea, particularly in developing nations.
- The EPEC effector protein EspF disrupts host intestinal barrier function and initiates mitochondrial cell death.
- The precise mechanisms by which EspF affects host cells remain incompletely understood.
Purpose of the Study:
- To identify host cell proteins that bind to the EPEC EspF effector.
- To elucidate the role of EspF-interacting proteins in EPEC-induced host cell death.
Main Methods:
- Affinity purification to identify EspF-binding proteins.
- Yeast two-hybrid assays, co-localization, and co-immunoprecipitation to confirm interactions.
- RNA interference (RNAi) to knock down Abcf2 expression.
- Caspase cleavage assays to assess apoptosis.
Main Results:
- The ABC-transporter family member Abcf2 was identified as an EspF-binding protein.
- EspF interaction with Abcf2 was confirmed through multiple experimental approaches.
- EPEC infection led to a dose-dependent decrease in Abcf2 levels.
- Abcf2 knockdown enhanced EspF-induced apoptosis and apoptosis induced by staurosporine.
Conclusions:
- EspF targets and interferes with the protective function of Abcf2.
- EspF induces or facilitates host cell death by disrupting Abcf2's role.
- Abcf2 may play a role in protecting host cells from apoptosis.
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