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Updated: May 22, 2025
Purification of Viral DNA for the Identification of Associated Viral and Cellular Proteins
Published on: August 31, 2017
Proteomic profiling of pseudorabies virus-infected PK-15 cells based on 4D label free analysis
Shuaiwei Wang1, Xiaoyong Chen2, Houqiang Luo1
1Department of Animal Science, Wenzhou Vocational College of Science and Technology, Wenzhou, China.
Abstract:
Pseudorabies virus (PRV) heavily depends on host machinery to support its life cycle. Investigating the interaction between PRV and host could aid in the understandings of viral pathogenesis. In this study, we performed a 4D label free proteomic method to examine the differentially expressed proteins in porcine kidney PK-15 cells with PRV infection. The results showed that the levels of 661 proteins were significantly elevated and 693 proteins were markedly reduced. Furthermore, these altered proteins were primarily enriched in spliceosome, protein processing in endoplasmic reticulum (ER), RNA transport, and protein export. To ensure the reliability of the proteomic results, the protein levels of formin binding protein 11 and wolfram syndrome 1as components of spliceosome and ER were verified via western blotting and the results were consistent. Together, our data shed light on a new protein profiling induced by PRV infection and highlighted the importance of spliceosome and ER in PRV replication which could promote understandings of host-PRV interplay.
Insights
This study reveals how Pseudorabies virus (PRV) alters host cell proteins, particularly in the spliceosome and endoplasmic reticulum (ER), impacting viral replication and pathogenesis.
Area of Science:
- Proteomics
- Virology
- Cell Biology
Background:
- Pseudorabies virus (PRV) relies on host cell machinery for its life cycle.
- Understanding host-pathogen interactions is crucial for elucidating viral pathogenesis.
Purpose of the Study:
- To investigate the proteomic changes in porcine kidney PK-15 cells upon PRV infection.
- To identify host proteins and pathways affected by PRV.
Main Methods:
- Utilized a 4D label-free proteomic approach.
- Analyzed differentially expressed proteins in PRV-infected PK-15 cells.
- Validated key protein changes using western blotting.
Main Results:
- Identified 661 upregulated and 693 downregulated proteins.
- Enrichment analysis revealed significant alterations in spliceosome, endoplasmic reticulum (ER) protein processing, RNA transport, and protein export pathways.
- Western blot confirmed the proteomic findings for formin binding protein 11 and wolfram syndrome 1.
Conclusions:
- PRV infection induces a distinct host protein profile.
- The spliceosome and ER are critical for PRV replication.
- Findings enhance understanding of the host-PRV interplay and viral pathogenesis.
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