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Investigation of the Cry4B-prohibitin interaction in Aedes aegypti cells
Atichat Kuadkitkan1, Duncan R Smith, Colin Berry
1Institute of Molecular Biosciences, Mahidol University, Salaya, Thailand.
Abstract:
Bacillus thuringiensis (Bt) produces insecticidal toxins active against insects. Cry4B, one of the major insecticidal toxins produced by Bt subsp. israelensis, is highly toxic to mosquitoes in the genus Aedes: the major vectors of dengue, yellow fever, and chikungunya. Previous work has shown that Cry4B binds to several mid-gut membrane proteins in Aedes aegypti larvae including prohibitin, a protein recently identified as a receptor that also mediates entry of dengue virus into Aedes cells. This study confirms the interaction between Cry4B and prohibitin by co-immunoprecipitation analysis and demonstrates colocalization of prohibitin and Cry4B by confocal microscopy. While activated Cry4B toxin showed high larvicidal activity, it was not cytotoxic to two Aedes cell lines, allowing determination of its effect on dengue virus infectivity in the absence of Cry4B-induced cell lysis. Pre-exposure of Aedes cells to Cry4B resulted in a significant reduction in the number of infected cells compared to untreated cells.
Insights
Bacillus thuringiensis (Bt) Cry4B toxin targets mosquito prohibitin receptors. This interaction reduces dengue virus infection in Aedes cells, offering potential for vector control.
Area of Science:
- Medical entomology
- Molecular biology
- Virology
Background:
- Bacillus thuringiensis (Bt) produces insecticidal toxins, including Cry4B, crucial for mosquito control.
- Cry4B targets mosquito mid-gut proteins, with prohibitin identified as a dengue virus entry receptor in Aedes aegypti.
- Understanding Cry4B-prohibitin interactions is key for developing novel dengue control strategies.
Purpose of the Study:
- To confirm the interaction between Bt Cry4B toxin and prohibitin in Aedes aegypti.
- To investigate the effect of Cry4B on dengue virus infectivity in Aedes cell lines.
Main Methods:
- Co-immunoprecipitation assays to verify Cry4B-prohibitin binding.
- Confocal microscopy to visualize protein colocalization.
- Assessing Cry4B larvicidal activity and cytotoxicity.
- Evaluating dengue virus infectivity in Cry4B-pre-exposed Aedes cells.
Main Results:
- Cry4B confirmed to interact with prohibitin.
- Colocalization of Cry4B and prohibitin demonstrated.
- Activated Cry4B showed high larvicidal activity but no cytotoxicity to Aedes cell lines.
- Pre-exposure to Cry4B significantly reduced dengue virus infection in Aedes cells.
Conclusions:
- Cry4B binds to prohibitin, a shared receptor for the toxin and dengue virus.
- Cry4B exhibits potential as an antiviral agent against dengue virus in Aedes mosquitoes.
- This finding opens avenues for integrated vector and virus control strategies.
