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Japanese encephalitis virus perturbs PML-nuclear bodies by engaging in interactions with distinct porcine PML
Songbai Yang1, Huaijin Liu1, Zhenyu Chen1
1Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Hangzhou, China.
Abstract:
Promyelocytic leukemia (PML) protein constitutes an indispensable element within PML-nuclear bodies (PML-NBs), playing a pivotal role in the regulation of multiple cellular functions while coordinating the innate immune response against viral invasions. Simultaneously, numerous viruses elude immune detection by targeting PML-NBs. Japanese encephalitis virus (JEV) is a flavivirus that causes Japanese encephalitis, a severe neurological disease that affects humans and animals. However, the mechanism through which JEV evades immunity via PML-NBs has been scarcely investigated. In the present study, PK15 cells were infected with JEV, and the quantity of intracellular PML-NBs was enumerated. The immunofluorescence results indicated that the number of PML-NBs was significantly reduced in JEV antigen-positive cells compared to viral antigen-negative cells. Subsequently, ten JEV proteins were cloned and transfected into PK15 cells. The results revealed that JEV non-structural proteins, NS2B, NS3, NS4A, NS4B, and NS5, significantly diminished the quantity of PML-NBs. Co-transfection was performed with the five JEV proteins and various porcine PML isoforms. The results demonstrated that NS2B colocalized with PML4 and PML5, NS4A colocalized with PML1 and PML4, NS4B colocalized with PML1, PML3, PML4, and PML5, while NS3 and NS5 interacted with all five PML isoforms. Furthermore, ectopic expression of PML isoforms confirmed that PML1, PML3, PML4, and PML5 inhibited JEV replication. These findings suggest that JEV disrupts the structure of PML-NBs through interaction with PML isoforms, potentially leading to the attenuation of the host's antiviral immune response.
Insights
Japanese encephalitis virus (JEV) infection reduces Promyelocytic leukemia protein nuclear bodies (PML-NBs). JEV non-structural proteins interact with PML isoforms, disrupting PML-NBs and potentially weakening the antiviral immune response.
Area of Science:
- Virology
- Cell Biology
- Immunology
Background:
- Promyelocytic leukemia (PML) protein and its nuclear bodies (PML-NBs) are crucial for cellular functions and innate immunity against viruses.
- Viruses often target PML-NBs to evade host immune detection.
- Japanese encephalitis virus (JEV) causes severe neurological disease, but its immune evasion mechanisms involving PML-NBs are poorly understood.
Purpose of the Study:
- To investigate how JEV interacts with PML-NBs to evade the host immune response.
- To identify specific JEV proteins involved in the disruption of PML-NBs.
- To explore the interaction between JEV proteins and PML isoforms and their effect on viral replication.
Main Methods:
- Infection of PK15 cells with JEV and enumeration of intracellular PML-NBs.
- Immunofluorescence assays to visualize PML-NBs and JEV antigen.
- Cloning and transfection of ten JEV proteins into PK15 cells.
- Co-transfection of JEV proteins with porcine PML isoforms.
- Assessment of JEV replication upon ectopic expression of PML isoforms.
Main Results:
- JEV infection significantly reduced the number of PML-NBs in infected cells.
- Five JEV non-structural proteins (NS2B, NS3, NS4A, NS4B, NS5) were found to significantly diminish PML-NB quantity.
- Specific JEV proteins showed distinct colocalization patterns with different PML isoforms (e.g., NS2B with PML4/PML5, NS3/NS5 with all isoforms).
- PML isoforms PML1, PML3, PML4, and PML5 inhibited JEV replication.
Conclusions:
- JEV disrupts PML-NB structure by interacting with various PML isoforms.
- This disruption likely contributes to the evasion of the host's antiviral immune response by JEV.
- Understanding these interactions provides insights into JEV pathogenesis and potential therapeutic targets.
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