Regulation of MDA5-MAVS Antiviral Signaling Axis by TRIM25 through TRAF6-Mediated NF-κB Activation

  • Lee, Na-Rae (Department of Pharmaceutical Science, College of Pharmacy, Kyung Hee University) ;
  • Kim, Hye-In (Department of Pharmaceutical Science, College of Pharmacy, Kyung Hee University) ;
  • Choi, Myung-Soo (Department of Pharmaceutical Science, College of Pharmacy, Kyung Hee University) ;
  • Yi, Chae-Min (Department of Pharmaceutical Science, College of Pharmacy, Kyung Hee University) ;
  • Inn, Kyung-Soo (Department of Pharmaceutical Science, College of Pharmacy, Kyung Hee University)
  • Received : 2015.03.17
  • Accepted : 2015.07.01
  • Published : 2015.09.30


Tripartite motif protein 25 (TRIM25), mediates K63-linked polyubiquitination of Retinoic acid inducible gene I (RIG-I) that is crucial for downstream antiviral interferon signaling. Here, we demonstrate that TRIM25 is required for melanoma differentiation-associated gene 5 (MDA5) and MAVS mediated activation of NF-${\kappa}B$ and interferon production. TRIM25 is required for the full activation of NF-${\kappa}B$ at the downstream of MAVS, while it is not involved in IRF3 nuclear translocation. Mechanical studies showed that TRIM25 is involved in TRAF6-mediated NF-${\kappa}B$ activation. These collectively indicate that TRIM25 plays an additional role in RIG-I/MDA5 signaling other than RIG-I ubiquitination via activation of NF-${\kappa}B$.


Supported by : National Research Foundation of Korea (NRF)


  1. Gack, M.U., Shin, Y.C., Joo, C.H., Urano, T., Liang, C., Sun, L., Takeuchi, O., Akira, S., Chen, Z., Inoue, S., et al. (2007). TRIM25 RING-finger E3 ubiquitin ligase is essential for RIG-Imediated antiviral activity. Nature 446, 916-920.
  2. Gack, M.U., Albrecht, R.A., Urano, T., Inn, K.S., Huang, I.C., Carnero, E., Farzan, M., Inoue, S., Jung, J.U., and Garcia-Sastre, A. (2009). Influenza A virus NS1 targets the ubiquitin ligase TRIM25 to evade recognition by the host viral RNA sensor RIG-I. Cell Host Microbe 5, 439-449.
  3. Goubau, D., Deddouche, S., and Reis, E.S.C. (2013). Cytosolic sensing of viruses. Immunity 38, 855-869.
  4. Inn, K.S., Gack, M.U., Tokunaga, F., Shi, M., Wong, L.Y., Iwai, K., and Jung, J.U. (2011a). Linear ubiquitin assembly complex negatively regulates RIG-I- and TRIM25-mediated type I interferon induction. Mol. Cell 41, 354-365.
  5. Inn, K.S., Lee, S.H., Rathbun, J.Y., Wong, L.Y., Toth, Z., Machida, K., Ou, J.H., and Jung, J.U. (2011b). Inhibition of RIG-I-mediated signaling by Kaposi's sarcoma-associated herpesvirus-encoded deubiquitinase ORF64. J. Virol. 85, 10899-10904.
  6. Jiang, X., Kinch, L.N., Brautigam, C.A., Chen, X., Du, F., Grishin, N.V., and Chen, Z.J. (2012). Ubiquitin-induced oligomerization of the RNA sensors RIG-I and MDA5 activates antiviral innate immune response. Immunity 36, 959-973.
  7. Kato, H., Takeuchi, O., Sato, S., Yoneyama, M., Yamamoto, M., Matsui, K., Uematsu, S., Jung, A., Kawai, T., Ishii, K.J., et al. (2006). Differential roles of MDA5 and RIG-I helicases in the recognition of RNA viruses. Nature 441, 101-105.
  8. Kato, H., Takeuchi, O., Mikamo-Satoh, E., Hirai, R., Kawai, T., Matsushita, K., Hiiragi, A., Dermody, T.S., Fujita, T., and Akira, S. (2008). Length-dependent recognition of double-stranded ribonucleic acids by retinoic acid-inducible gene-I and melanoma differentiation-associated gene 5. J. Exp. Med. 205, 1601-1610.
  9. Kawai, T., Takahashi, K., Sato, S., Coban, C., Kumar, H., Kato, H., Ishii, K.J., Takeuchi, O., and Akira, S. (2005). IPS-1, an adaptor triggering RIG-I- and Mda5-mediated type I interferon induction. Nat. Immunol. 6, 981-988.
  10. Loo, Y.M., Fornek, J., Crochet, N., Bajwa, G., Perwitasari, O., Martinez-Sobrido, L., Akira, S., Gill, M.A., Garcia-Sastre, A., Katze, M.G., et al. (2008). Distinct RIG-I and MDA5 signaling by RNA viruses in innate immunity. J. Virol. 82, 335-345.
  11. Meylan, E., Curran, J., Hofmann, K., Moradpour, D., Binder, M., Bartenschlager, R., and Tschopp, J. (2005). Cardif is an adaptor protein in the RIG-I antiviral pathway and is targeted by hepatitis C virus. Nature 437, 1167-1172.
  12. Nakhaei, P., Genin, P., Civas, A., and Hiscott, J. (2009). RIG-I-like receptors: sensing and responding to RNA virus infection. Semin. Immunol. 21, 215-222.
  13. Pichlmair, A., Schulz, O., Tan, C.P., Rehwinkel, J., Kato, H., Takeuchi, O., Akira, S., Way, M., Schiavo, G., and Reis e Sousa, C. (2009). Activation of MDA5 requires higher-order RNA structures generated during virus infection. J. Virol. 83, 10761-10769.
  14. Ramos, H.J., and Gale, M., Jr. (2011). RIG-I like receptors and their signaling crosstalk in the regulation of antiviral immunity. Curr. Opin. Virol. 1, 167-176.
  15. Seth, R.B., Sun, L., Ea, C.K., and Chen, Z.J. (2005). Identification and characterization of MAVS, a mitochondrial antiviral signaling protein that activates NF-kappaB and IRF 3. Cell 122, 669-682.
  16. Sumpter, R., Jr., Loo, Y.M., Foy, E., Li, K., Yoneyama, M., Fujita, T., Lemon, S.M., and Gale, M., Jr. (2005). Regulating intracellular antiviral defense and permissiveness to hepatitis C virus RNA replication through a cellular RNA helicase, RIG-I. J. Virol. 79, 2689-2699.
  17. Wies, E., Wang, M.K., Maharaj, N.P., Chen, K., Zhou, S., Finberg, R.W., and Gack, M.U. (2013). Dephosphorylation of the RNA sensors RIG-I and MDA5 by the phosphatase PP1 is essential for innate immune signaling. Immunity 38, 437-449.
  18. Wu, B., Peisley, A., Richards, C., Yao, H., Zeng, X., Lin, C., Chu, F., Walz, T., and Hur, S. (2013). Structural basis for dsRNA recognition, filament formation, and antiviral signal activation by MDA5. Cell 152, 276-289.
  19. Xu, L.G., Wang, Y.Y., Han, K.J., Li, L.Y., Zhai, Z., and Shu, H.B. (2005). VISA is an adapter protein required for virus-triggered IFN-beta signaling. Mol. Cell 19, 727-740.
  20. Yoboua, F., Martel, A., Duval, A., Mukawera, E., and Grandvaux, N. (2010). Respiratory syncytial virus-mediated NF-kappa B p65 phosphorylation at serine 536 is dependent on RIG-I, TRAF6, and IKK beta. J. Virol. 84, 7267-7277.
  21. Yoneyama, M., Kikuchi, M., Natsukawa, T., Shinobu, N., Imaizumi, T., Miyagishi, M., Taira, K., Akira, S., and Fujita, T. (2004). The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses. Nat. Immunol. 5, 730-737.
  22. Yoshida, R., Takaesu, G., Yoshida, H., Okamoto, F., Yoshioka, T., Choi, Y., Akira, S., Kawai, T., Yoshimura, A., and Kobayashi, T. (2008). TRAF6 and MEKK1 play a pivotal role in the RIG-I-like helicase antiviral pathway. J. Biol. Chem. 283, 36211-36220.
  23. Zeng, W., Sun, L., Jiang, X., Chen, X., Hou, F., Adhikari, A., Xu, M., and Chen, Z.J. (2010). Reconstitution of the RIG-I pathway reveals a signaling role of unanchored polyubiquitin chains in innate immunity. Cell 141, 315-330.

Cited by

  1. TRIM25 in the Regulation of the Antiviral Innate Immunity vol.8, 2017,
  2. Molecular cloning and functional analysis of tumor necrosis factor receptor-associated factor 6 (TRAF6) in Crossastrea gigas vol.68, 2017,
  3. De novo annotation of the immune-enriched transcriptome provides insights into immune system genes of Chinese sturgeon (Acipenser sinensis) vol.55, 2016,
  4. Ubiquitin in the activation and attenuation of innate antiviral immunity vol.213, pp.1, 2016,
  5. The Severe Acute Respiratory Syndrome Coronavirus Nucleocapsid Inhibits Type I Interferon Production by Interfering with TRIM25-Mediated RIG-I Ubiquitination vol.91, pp.8, 2017,
  6. Herpesvirus deconjugases inhibit the IFN response by promoting TRIM25 autoubiquitination and functional inactivation of the RIG-I signalosome vol.14, pp.1, 2018,
  7. TRIM25 enhances cell growth and cell survival by modulating p53 signals via interaction with G3BP2 in prostate cancer vol.37, pp.16, 2018,
  8. USP4 positively regulates RLR-induced NF-κB activation by targeting TRAF6 for K48-linked deubiquitination and inhibits enterovirus 71 replication vol.8, pp.1, 2018,
  9. Gene in Chinese Patients with Dyschromatosis Symmetrica Hereditaria vol.22, pp.2, 2018,