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| Screening of VISA Interacting Proteins by Yeast Two-hybrid System |
| WANG Dan-dan1,2, CHEN Tian1,2, XU Liang-guo1,2 |
1. Key Laboratory of Small Functional Organic Molecule, Ministry of Education, Jiangxi Normal University, Nanchang 330022, China;
2. College of Life Sciences, Jiangxi Normal University, Nanchang 330022, China |
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Abstract The infection of a virus is a great threat to host; the innate immune system is set up as the natural protection system of an organism. VISA (also know as MAVS, CARDIF, IPS-1), is an critical adapter protein required for virus-triggered IFN-β signaling, recruiting RIG-I-like receptors(including RIG-I and MDA5) and downstream signaling proteins to VISA signalosome at mitochondria upon virus infection, leading to activation of transcriptional factor IRF3 and NF-κB, and resulting in the induction of IFNs and other inflammatory cytokines. However, the mystery of VISA for the antiviral function is still not completely figured out. VISA was used as bait for screening a human 293T cell cDNA library in a yeast two-hybrid screening, trying to identify the interacting proteins of VISA. Then, coimmunoprecipitation experiments were carried out to verify the association between VISA and candidates proteins screened from the yeast two-hybrid system assay. It is indicated that Sec13L1 associated with VISA. Overexpression of Sec13L1 in 293 cells facilitated to enhance the activation of IFNβ promoter induced by VISA and virus infection in a dose-dependent manner.
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Received: 27 February 2017
Published: 25 June 2017
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[5] Moreira T G, Zhang L, Shaulovet L, et al. Sec13 regulates expression of specific immune factors involved in inflammation in vivo. Scientific Reports, 2015, 5:17655.
[6] He Y, Yang F, Wang F, et al. The upregulation of expressed proteins in HepG2 cells transfected by the recombinant plasmid-containing HBx gene. Scandinavian Journal of Immunology, 2007, 65(3):249-256. |
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