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Updated: Oct 15, 2025
Analysis of Yersinia enterocolitica Effector Translocation into Host Cells Using Beta-lactamase Effector Fusions
Published on: October 13, 2015
Bimolecular fluorescence complementation assay to explore protein-protein interactions of the Yersinia virulence
1Ege University, Faculty of Science, Department of Biochemistry, 35100, Bornova-Izmir, Turkey; Department of Medical Services and Techniques, Hatay Vocational School of Health Services, Hatay Mustafa Kemal University, Tayfur Sökmen Campus, 31060, Alahan-Antakya/ Hatay, Turkey.
Abstract:
Yersinia outer protein M (YopM) is one of the effector proteins and essential for virulence. YopM is delivered by the Yersinia type III secretion system (T3SS) into the host cell, where it shows immunosuppressive effect through interaction with host proteins. Therefore, protein-protein interactions of YopM is significant to understand its molecular mechanism. In this study, we aimed to explore protein-protein interactions of YopM with the two components of T3SS, namely LcrV and LcrG. We used bimolecular fluorescence complementation (BiFC) assay and monitored the reassembly of green fluorescence protein in Escherichia coli. As an indicator of the protein-protein interaction, we monitored the in vivo reconstitution of fluorescence by measuring fluorescence intensity and imaging the cells under fluorescence microscope. We showed, for the first time, that YopM interacts with LcrG, but not with LcrV. Here, we propose BiFC assay as a simple method to screen novel interaction partners of YopM.
Insights
Yersinia outer protein M (YopM) interacts with LcrG, a component of the type III secretion system (T3SS), but not LcrV. This finding, revealed by a bimolecular fluorescence complementation assay, advances understanding of YopM's virulence mechanism.
Area of Science:
- Microbiology
- Molecular Biology
- Immunology
Background:
- Yersinia outer protein M (YopM) is a key virulence effector protein in Yersinia species.
- YopM is translocated into host cells via the type III secretion system (T3SS), mediating immunosuppression through host protein interactions.
- Understanding YopM's protein-protein interactions is crucial for elucidating its molecular mechanisms of virulence.
Purpose of the Study:
- To investigate the in vivo interactions between Yersinia outer protein M (YopM) and two T3SS components, LcrV and LcrG.
- To establish the bimolecular fluorescence complementation (BiFC) assay as a method for studying YopM interactions.
Main Methods:
- Utilized the bimolecular fluorescence complementation (BiFC) assay in Escherichia coli.
- Monitored in vivo fluorescence reconstitution by measuring fluorescence intensity and imaging.
- Assessed YopM interactions with LcrV and LcrG.
Main Results:
- Demonstrated a direct interaction between YopM and LcrG.
- Showed no detectable interaction between YopM and LcrV.
- Validated the BiFC assay for detecting YopM protein-protein interactions.
Conclusions:
- Yersinia outer protein M (YopM) interacts with LcrG, a critical finding for understanding T3SS function.
- The study highlights the utility of the BiFC assay for screening YopM interaction partners.
- This research contributes to the understanding of Yersinia pathogenesis and immune evasion strategies.
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