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Generation of Monoclonal Cultures from Wolbachia-infected Drosophila melanogaster JW18 Cell Line
Published on: June 27, 2025
507
Differential viral RNA methylation contributes to pathogen blocking in Wolbachia-colonized arthropods
Tamanash Bhattacharya1,2, Liewei Yan3, John M Crawford1
1Department of Biology, Indiana University, Bloomington, Indiana, United States of America.
Plos Pathogens
|March 16, 2022
Summary
Wolbachia bacteria suppress alphavirus replication in mosquitoes by reducing the activity of the DNMT2 enzyme. This epigenetic modification disruption hinders viral RNA replication and transmission, aiding biocontrol efforts.
Area of Science:
- Virology
- Molecular Biology
- Entomology
Background:
- Wolbachia pipientis is an endosymbiont used for biocontrol of mosquito-borne viruses.
- Host cytosine methyltransferase DNMT2 was previously identified as crucial for pathogen-blocking in Drosophila.
- The role of DNMT2 in Wolbachia-mediated virus inhibition in Aedes mosquitoes was unexplored.
Purpose of the Study:
- To investigate the role of DNMT2 in Wolbachia-induced alphavirus inhibition in Aedes species.
- To determine if DNMT2 activity affects viral RNA modification and replication in mosquitoes.
- To elucidate the mechanism by which Wolbachia suppresses alphavirus.
Main Methods:
- Assessing DNMT2 expression in virus-infected and Wolbachia-colonized Aedes mosquitoes.
- Measuring viral replication and progeny virus infectivity in mosquito tissues and cell cultures.
- Utilizing ectopic expression of DNMT2 in cultured Aedes cells.
- Employing Liquid Chromatography-Mass Spectrometry (LC-MS) to analyze viral RNA modifications.
Main Results:
- DNMT2 expression increased upon viral infection but was suppressed in Wolbachia-colonized mosquitoes.
- Wolbachia colonization correlated with reduced viral replication and progeny virus infectivity.
- Ectopic DNMT2 expression in Aedes cells promoted viral replication and infectivity, dependent on its methyltransferase activity.
- LC-MS analysis revealed decreased 5-methylcytosine modification on viral RNA in Wolbachia-colonized cells and animals.
Conclusions:
- DNMT2 plays a proviral role in alphavirus replication and transmission in Aedes mosquitoes.
- Wolbachia suppresses alphavirus by down-regulating DNMT2, leading to reduced 5-methylcytosine modification of viral RNA.
- Disruption of viral RNA epitranscriptomic modifications is a key mechanism in Wolbachia-mediated pathogen blocking.
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