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

Lee NR, Kim HI, Choi MS, Yi CM, Inn KS - Mol. Cells (2015)

Bottom Line: 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.Mechanical studies showed that TRIM25 is involved in TRAF6-mediated NF-κB activation.These collectively indicate that TRIM25 plays an additional role in RIG-I/MDA5 signaling other than RIG-I ubiquitination via activation of NF-κB.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Science, College of Pharmacy, Kyung Hee University, Seoul 130-701, Korea.

ABSTRACT
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-κB and interferon production. TRIM25 is required for the full activation of NF-κB 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-κB activation. These collectively indicate that TRIM25 plays an additional role in RIG-I/MDA5 signaling other than RIG-I ubiquitination via activation of NF-κB.

No MeSH data available.


Related in: MedlinePlus

Role of TRIM25 in MAVS-mediated signaling. (A) TRIM25 is required for efficient activation of IFN-β promoter by MAVS. Wild-type (WT) and TRIM25 knock-out (TRIM25 KO) MEFs were transfected with vector or MAVS plasmids (hMAVS). Cells were co-transfected with IFN-β promoter-reporter and TK-Renilla reporter plasmids. Promoter activities were determined by Dual-Luciferase assay 16 h after transfection. (B) TRIM25 is required for efficient activation of NF-κB promoter by MAVS. WT and TRIM25 KO MEFs were transfected with vector or MAVS plasmids together with NF-κB reporter and TK-Renilla reporter plasmids. Promoter activities were determined using procedures similar to those in (A). (C) Enhanced MAVS-induced IFN-β promoter activity by TRIM25 overexpression. HEK293T cells were transfected with vector, MAVS and TRIM25 plasmids as indicated, together with IFN-β reporter and TK-Renilla reporter plasmids. Promoter activities were determined using procedures similar to those in (A). (D) Enhanced MAVS-induced NF-κB promoter activity by TRIM25 overexpression. HEK293T cells were transfected with expression plasmids and NF-κB reporter plasmids as indicated. Promoter activities were determined using procedures similar to those in (C). Results of experiments, performed in triplicate, are presented as means ± standard deviation.
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f2-molce-38-9-759: Role of TRIM25 in MAVS-mediated signaling. (A) TRIM25 is required for efficient activation of IFN-β promoter by MAVS. Wild-type (WT) and TRIM25 knock-out (TRIM25 KO) MEFs were transfected with vector or MAVS plasmids (hMAVS). Cells were co-transfected with IFN-β promoter-reporter and TK-Renilla reporter plasmids. Promoter activities were determined by Dual-Luciferase assay 16 h after transfection. (B) TRIM25 is required for efficient activation of NF-κB promoter by MAVS. WT and TRIM25 KO MEFs were transfected with vector or MAVS plasmids together with NF-κB reporter and TK-Renilla reporter plasmids. Promoter activities were determined using procedures similar to those in (A). (C) Enhanced MAVS-induced IFN-β promoter activity by TRIM25 overexpression. HEK293T cells were transfected with vector, MAVS and TRIM25 plasmids as indicated, together with IFN-β reporter and TK-Renilla reporter plasmids. Promoter activities were determined using procedures similar to those in (A). (D) Enhanced MAVS-induced NF-κB promoter activity by TRIM25 overexpression. HEK293T cells were transfected with expression plasmids and NF-κB reporter plasmids as indicated. Promoter activities were determined using procedures similar to those in (C). Results of experiments, performed in triplicate, are presented as means ± standard deviation.

Mentions: Upon activation of RIG-I and MDA5, both are recruited to MAVS, a critical downstream molecule. The effect of TRIM25 on MAVS-induced interferon signaling was investigated to determine whether TRIM25 acts upstream or downstream of MAVS. MAVS-induced interferon-β promoter activity and NF-κB transcriptional activity in TRIM25 deficient MEFs were lower than those in wild-type MEFs, indicating that TRIM25 may act downstream of MAVS (Figs. 2A and 2B). In addition, ectopic expression of TRIM25 augmented MAVS-induced activation of interferon-β promoter activity and NF-κB transcriptional activities (Figs. 2C and 2D).


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

Lee NR, Kim HI, Choi MS, Yi CM, Inn KS - Mol. Cells (2015)

Role of TRIM25 in MAVS-mediated signaling. (A) TRIM25 is required for efficient activation of IFN-β promoter by MAVS. Wild-type (WT) and TRIM25 knock-out (TRIM25 KO) MEFs were transfected with vector or MAVS plasmids (hMAVS). Cells were co-transfected with IFN-β promoter-reporter and TK-Renilla reporter plasmids. Promoter activities were determined by Dual-Luciferase assay 16 h after transfection. (B) TRIM25 is required for efficient activation of NF-κB promoter by MAVS. WT and TRIM25 KO MEFs were transfected with vector or MAVS plasmids together with NF-κB reporter and TK-Renilla reporter plasmids. Promoter activities were determined using procedures similar to those in (A). (C) Enhanced MAVS-induced IFN-β promoter activity by TRIM25 overexpression. HEK293T cells were transfected with vector, MAVS and TRIM25 plasmids as indicated, together with IFN-β reporter and TK-Renilla reporter plasmids. Promoter activities were determined using procedures similar to those in (A). (D) Enhanced MAVS-induced NF-κB promoter activity by TRIM25 overexpression. HEK293T cells were transfected with expression plasmids and NF-κB reporter plasmids as indicated. Promoter activities were determined using procedures similar to those in (C). Results of experiments, performed in triplicate, are presented as means ± standard deviation.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4588718&req=5

f2-molce-38-9-759: Role of TRIM25 in MAVS-mediated signaling. (A) TRIM25 is required for efficient activation of IFN-β promoter by MAVS. Wild-type (WT) and TRIM25 knock-out (TRIM25 KO) MEFs were transfected with vector or MAVS plasmids (hMAVS). Cells were co-transfected with IFN-β promoter-reporter and TK-Renilla reporter plasmids. Promoter activities were determined by Dual-Luciferase assay 16 h after transfection. (B) TRIM25 is required for efficient activation of NF-κB promoter by MAVS. WT and TRIM25 KO MEFs were transfected with vector or MAVS plasmids together with NF-κB reporter and TK-Renilla reporter plasmids. Promoter activities were determined using procedures similar to those in (A). (C) Enhanced MAVS-induced IFN-β promoter activity by TRIM25 overexpression. HEK293T cells were transfected with vector, MAVS and TRIM25 plasmids as indicated, together with IFN-β reporter and TK-Renilla reporter plasmids. Promoter activities were determined using procedures similar to those in (A). (D) Enhanced MAVS-induced NF-κB promoter activity by TRIM25 overexpression. HEK293T cells were transfected with expression plasmids and NF-κB reporter plasmids as indicated. Promoter activities were determined using procedures similar to those in (C). Results of experiments, performed in triplicate, are presented as means ± standard deviation.
Mentions: Upon activation of RIG-I and MDA5, both are recruited to MAVS, a critical downstream molecule. The effect of TRIM25 on MAVS-induced interferon signaling was investigated to determine whether TRIM25 acts upstream or downstream of MAVS. MAVS-induced interferon-β promoter activity and NF-κB transcriptional activity in TRIM25 deficient MEFs were lower than those in wild-type MEFs, indicating that TRIM25 may act downstream of MAVS (Figs. 2A and 2B). In addition, ectopic expression of TRIM25 augmented MAVS-induced activation of interferon-β promoter activity and NF-κB transcriptional activities (Figs. 2C and 2D).

Bottom Line: 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.Mechanical studies showed that TRIM25 is involved in TRAF6-mediated NF-κB activation.These collectively indicate that TRIM25 plays an additional role in RIG-I/MDA5 signaling other than RIG-I ubiquitination via activation of NF-κB.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Science, College of Pharmacy, Kyung Hee University, Seoul 130-701, Korea.

ABSTRACT
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-κB and interferon production. TRIM25 is required for the full activation of NF-κB 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-κB activation. These collectively indicate that TRIM25 plays an additional role in RIG-I/MDA5 signaling other than RIG-I ubiquitination via activation of NF-κB.

No MeSH data available.


Related in: MedlinePlus