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VopE, a Vibrio cholerae Type III Effector, Attenuates the Activation of CWI-MAPK Pathway in Yeast Model System
Leela K Bankapalli1, Rahul C Mishra1, Saumya Raychaudhuri1
1Molecular Biology and Microbial Physiology, Institute of Microbial Technology Chandigarh, India.
Abstract:
VopE, a mitochondrial targeting T3SS effector protein of Vibrio cholerae, perturbs innate immunity by modulating mitochondrial dynamics. In the current study, ectopic expression of VopE was found to be toxic in a yeast model system and toxicity was further aggravated in the presence of various stressors. Interestingly, a VopE variant lacking predicted mitochondrial targeting sequence (MTS) also exhibited partial lethality in the yeast system. With the aid of yeast genetic tools and different stressors, we have demonstrated that VopE and its derivative VopEΔMTS modulate cell wall integrity (CWI-MAPK) signaling pathway and have identified several critical residues contributing to the lethality of VopE. Furthermore, co-expression of two effectors VopEΔMTS and VopX, interfering with the CWI-MAPK cellular pathway can partially suppress the VopX mediated yeast growth inhibition. Taken together, these results suggest that VopE alters signaling through the CWI-MAPK pathway, and demonstrates the usefulness of yeast model system to gain additional insights on the functionality of VopE.
Insights
Vibrio cholerae VopE protein disrupts innate immunity. Researchers found VopE and a variant (VopEΔMTS) impact yeast cell wall integrity signaling, revealing key residues and demonstrating yeast as a model for studying VopE function.
Area of Science:
- Microbiology
- Cell Biology
- Molecular Biology
Background:
- Vibrio cholerae secretes VopE, a type III secretion system (T3SS) effector.
- VopE targets mitochondria and modulates mitochondrial dynamics, impacting innate immunity.
- Understanding VopE's cellular functions is crucial for comprehending V cholerae pathogenesis.
Purpose of the Study:
- To investigate the toxicity and cellular effects of VopE in a yeast model system.
- To elucidate the role of the mitochondrial targeting sequence (MTS) in VopE toxicity.
- To identify the specific cellular pathways affected by VopE and its variants.
Main Methods:
- Ectopic expression of VopE and a VopE variant lacking MTS (VopEΔMTS) in yeast.
- Assessment of VopE toxicity under various stress conditions.
- Utilizing yeast genetic tools to analyze the cell wall integrity (CWI-MAPK) signaling pathway.
- Investigating the effects of co-expressing VopEΔMTS with VopX.
Main Results:
- Ectopic expression of VopE was toxic in yeast, exacerbated by stressors.
- VopEΔMTS also induced partial lethality, indicating MTS-independent effects.
- Both VopE and VopEΔMTS were shown to modulate the CWI-MAPK signaling pathway.
- Critical residues contributing to VopE lethality were identified.
- Co-expression of VopEΔMTS and VopX partially suppressed VopX-mediated growth inhibition.
Conclusions:
- VopE perturbs cellular signaling, specifically impacting the CWI-MAPK pathway.
- The yeast model system is effective for studying VopE's functional mechanisms.
- Mitochondrial targeting is not solely responsible for all observed VopE effects.