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Published on: December 29, 2015
Proteomic landscape of Japanese encephalitis virus-infected fibroblasts
Kiran Bala Sharma1,2, Simran Chhabra1, Suruchi Aggarwal2
1Regional Centre for Biotechnology, NCR Biotech Science Cluster, Faridabad, Haryana, India.
Abstract:
Advances in proteomics have enabled a comprehensive understanding of host-pathogen interactions. Here we have characterized Japanese encephalitis virus (JEV) infection-driven changes in the mouse embryonic fibroblast (MEF) proteome. Through tandem mass tagging (TMT)-based mass spectrometry, we describe changes in 7.85 % of the identified proteome due to JEV infection. Pathway enrichment analysis showed that proteins involved in innate immune sensing, interferon responses and inflammation were the major upregulated group, along with the immunoproteasome and poly ADP-ribosylation proteins. Functional validation of several upregulated anti-viral innate immune proteins, including an active cGAS-STING axis, was performed. Through siRNA depletion, we describe a crucial role of the DNA sensor cGAS in restricting JEV replication. Further, many interferon-stimulated genes (ISGs) were observed to be induced in infected cells. We also observed activation of TLR2 and inhibition of TLR2 signalling using TLR1/2 inhibitor CU-CPT22-blocked production of inflammatory cytokines IL6 and TNF-α from virus-infected N9 microglial cells. The major proteins that were downregulated by infection were involved in cell adhesion (collagens), transport (solute carrier and ATP-binding cassette transporters), sterol and lipid biosynthesis. Several collagens were found to be transcriptionally downregulated in infected MEFs and mouse brain. Collectively, our data provide a bird's-eye view into how fibroblast protein composition is rewired following JEV infection.
Insights
Japanese encephalitis virus (JEV) infection significantly alters the mouse fibroblast proteome, upregulating innate immune responses and downregulating cell adhesion proteins. This study reveals key host-pathogen interactions impacting viral restriction.
Area of Science:
- Virology
- Proteomics
- Immunology
Background:
- Host-pathogen interactions are crucial for understanding viral pathogenesis.
- Proteomics offers a powerful approach to study cellular responses during infection.
Purpose of the Study:
- To characterize Japanese encephalitis virus (JEV)-induced proteome changes in mouse embryonic fibroblasts (MEFs).
- To elucidate the host's innate immune response and cellular alterations during JEV infection.
Main Methods:
- Tandem mass tagging (TMT)-based mass spectrometry was employed to analyze proteome-wide changes.
- siRNA depletion was used to validate the role of specific proteins, such as cGAS.
- Pathway enrichment analysis identified key biological processes affected by JEV infection.
Main Results:
- JEV infection altered 7.85% of the identified proteome, with significant upregulation of innate immune sensing, interferon responses, and inflammation-related proteins.
- The cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway was activated, and cGAS was found to restrict JEV replication.
- Downregulated proteins were primarily involved in cell adhesion (collagens) and transport processes, with evidence of transcriptional downregulation in infected cells and brain tissue.
Conclusions:
- JEV infection profoundly rewires the fibroblast proteome, enhancing antiviral innate immunity while suppressing cellular functions like adhesion and transport.
- The cGAS-STING pathway plays a critical role in restricting JEV replication.
- These findings provide comprehensive insights into host responses to JEV infection at the proteomic level.
