Related Experiment Video

Updated: Oct 17, 2025

Analysis of Group IV Viral SSHHPS Using In Vitro and In Silico Methods
10:40

Analysis of Group IV Viral SSHHPS Using In Vitro and In Silico Methods

Published on: December 21, 2019

26.1K

Porcine TRIM35 positively regulate TRAF3-mediated IFN-β production and inhibit Japanese encephalitis virus

Chenxi Li1, Yanyang Zhou2, Xuan Chen3

  • 1College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009, Jiangsu, China; Comparative Medicine Research Institute, Yangzhou University, Yangzhou, 225009, Jiangsu, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, 225009, Jiangsu, China; State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, Harbin, 150069, China.

Developmental and Comparative Immunology
|October 9, 2021
PubMed

Insights

Porcine TRIM35 (porTRIM35) protein enhances interferon-beta (IFN-β) production by regulating ubiquitination, boosting antiviral immunity against Japanese encephalitis virus (JEV). This study details porTRIM35

Area of Science:

  • Immunology
  • Molecular Biology
  • Virology

Background:

  • Tripartite motif 35 (TRIM35) is a ubiquitin E3 ligase crucial for innate immune signaling and interferon-beta (IFN-β) production.
  • Understanding TRIM35's role in antiviral responses, particularly against economically significant viruses like Japanese encephalitis virus (JEV), is vital for livestock health.

Purpose of the Study:

  • To clone and characterize the porcine TRIM35 (porTRIM35) gene.
  • To investigate the role of porTRIM35 in IFN-β expression and antiviral defense against JEV.
  • To elucidate the molecular mechanism underlying porTRIM35-mediated IFN-β production.

Main Methods:

  • Gene cloning and sequence analysis of porTRIM35.
  • Quantitative PCR to assess mRNA expression levels of IFN-β.
  • Western blotting and co-immunoprecipitation to study protein interactions and ubiquitination.
  • Poly(I:C) stimulation and JEV infection in swine testicular (ST) cells.
  • siRNA-mediated knockdown of endogenous porTRIM35.

Main Results:

  • The porTRIM35 gene encodes a 493-amino acid protein with conserved TRIM structural domains.
  • Overexpression of porTRIM35 up-regulated IFN-β mRNA levels in response to poly(I:C), while knockdown decreased it.
  • porTRIM35 directly interacted with porcine TNF-receptor associated factor 3 (TRAF3) and mediated its Lys63-linked polyubiquitination, enhancing IFN-β production.
  • The RING and PRY/SPRY domains are essential for porTRIM35's E3 ligase activity.
  • porTRIM35 expression was inhibited by JEV infection, but its exogenous expression reduced viral titers and increased IFN-β production during infection.

Conclusions:

  • Porcine TRIM35 plays a significant role in regulating IFN-β production through TRAF3 ubiquitination.
  • porTRIM35 enhances the innate immune response against Japanese encephalitis virus (JEV) infection.
  • TRIM35 acts as a critical component in the antiviral defense mechanisms of swine.
Keywords:
Japanese encephalitis virusPorcine tripartite motif 35TNF-Receptor associated factor 3Type I interferonViral replication

More Related Videos

A High Resolution Method to Monitor Phosphorylation-dependent Activation of IRF311:44

A High Resolution Method to Monitor Phosphorylation-dependent Activation of IRF3

Published on: January 24, 2016

12.1K
Optimized Interferon-gamma ELISpot Assay to Measure T Cell Responses in the Guinea Pig Model after Vaccination08:13

Optimized Interferon-gamma ELISpot Assay to Measure T Cell Responses in the Guinea Pig Model after Vaccination

Published on: January 20, 2019

22.0K

Related Concept Videos

TGF - β Signaling Pathway01:16

TGF - β Signaling Pathway

The TGF-β signaling pathway regulates cell growth, differentiation, adhesion, motility, and development. TGF-β ligands that induce TGF-β signaling are synthesized in their latent form. Several proteases or cell surface receptors such as integrins act upon the latent form, releasing the active ligand. There are three types of mammalian TGF-βs: (TGF-β1, TGF-β2, and TGF-β3) that bind as homodimers or heterodimers to TGF-β receptors. The TGF-β receptors...
8.0K
Regulation of the Unfolded Protein Response01:31

Regulation of the Unfolded Protein Response

Inositol-requiring kinase one or IRE1 is the most conserved eukaryotic unfolded protein response (UPR) receptor. It is a type I transmembrane protein kinase receptor with a distinctive site-specific RNase activity. As the binding mechanics of the misfolded proteins with the N-terminal domain of IRE-1 are unclear, three binding models — direct, indirect, and allosteric -- are proposed for receptor activation. Nevertheless, it is known that once a misfolded protein associates with IRE1, it...
2.7K