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TRIM52 inhibits Japanese Encephalitis Virus replication by degrading the viral NS2A

View Article: PubMed Central - PubMed

ABSTRACT

The members of tripartite-motif containing (TRIM) protein participate in various cellular processes and play an important role in host antiviral function. TRIM proteins exert their antiviral activity either directly by degrading viral proteins through their E3 ligase activity, or indirectly by promoting host innate immunity. This study demonstrated for the first time that TRIM52 is a novel antiviral TRIM protein against Japanese encephalitis virus (JEV) infection. Overexpression of TRIM52 restricted JEV replication in BHK-21 and 293T cells. In addition, JEV nonstructural protein 2A (NS2A) is a protein that interacts with TRIM52. Their interaction degraded NS2A in a proteasome-dependent manner via the E3 ligase activity of TRIM52. Thus, TRIM52 is a novel antiviral TRIM protein, and it exerted antiviral activity against JEV infection by targeting and degrading viral NS2A.

No MeSH data available.


Related in: MedlinePlus

RING domain of TRIM52 is required for its antiviral activity against JEV replication.The 293T cells in 12-well plates were transfected with plasmid expressing full length TRIM52, RING domain-deleted TRIM52 and empty vector for 30 h and then infected with JEV (MOI = 1). The cells and culture supernatants were collected at 24 hpi. (a) Production of progeny virus in supernatants was determined using plaques assay. (b) Intracellular JEV NS3 protein was detected via Western blot analysis using the indicated antibodies. GAPDH was used to normalize the total protein loading. ***P < 0.001.
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f6: RING domain of TRIM52 is required for its antiviral activity against JEV replication.The 293T cells in 12-well plates were transfected with plasmid expressing full length TRIM52, RING domain-deleted TRIM52 and empty vector for 30 h and then infected with JEV (MOI = 1). The cells and culture supernatants were collected at 24 hpi. (a) Production of progeny virus in supernatants was determined using plaques assay. (b) Intracellular JEV NS3 protein was detected via Western blot analysis using the indicated antibodies. GAPDH was used to normalize the total protein loading. ***P < 0.001.

Mentions: TRIM52 is a RING-type E3 ligase and it can mediate NS2A degradation via the ubiquitin-proteasome pathway. We thus speculated that the RING domain of TRIM52 also contributes to the antiviral activity of TRIM52. To validate this conjecture, we transfected 293T cells with plasmids encoding either the full-length TRIM52or RING domain deleted TRIM52 (TRIM52dR) or with empty vector 30 h prior to JEV infection at an MOI of 1. Progeny virus in culture supernatants was collected at 24 hpi, and viral titers were determined by plaque assay. The cells with full-length TRIM52 showed an obviously reduced viral production compared with the cells with TRIM52dR and empty vector control (Fig. 6a). By contrast, viral production in cells with TRIM52dR versus empty vector control did not vary (Fig. 6a). Moreover, JEV NS3 protein expression was significantly reduced in cells overexpressing the full-length TRIM52 compared with cells overexpressing TRIM52dR and vector control (Fig. 6b). No difference was observed between cells with TRIM52dR and empty vector (Fig. 6b). Overall, these results suggest that the RING domain of TRIM52 is required for TRIM52 to control JEV replication.


TRIM52 inhibits Japanese Encephalitis Virus replication by degrading the viral NS2A
RING domain of TRIM52 is required for its antiviral activity against JEV replication.The 293T cells in 12-well plates were transfected with plasmid expressing full length TRIM52, RING domain-deleted TRIM52 and empty vector for 30 h and then infected with JEV (MOI = 1). The cells and culture supernatants were collected at 24 hpi. (a) Production of progeny virus in supernatants was determined using plaques assay. (b) Intracellular JEV NS3 protein was detected via Western blot analysis using the indicated antibodies. GAPDH was used to normalize the total protein loading. ***P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5035999&req=5

f6: RING domain of TRIM52 is required for its antiviral activity against JEV replication.The 293T cells in 12-well plates were transfected with plasmid expressing full length TRIM52, RING domain-deleted TRIM52 and empty vector for 30 h and then infected with JEV (MOI = 1). The cells and culture supernatants were collected at 24 hpi. (a) Production of progeny virus in supernatants was determined using plaques assay. (b) Intracellular JEV NS3 protein was detected via Western blot analysis using the indicated antibodies. GAPDH was used to normalize the total protein loading. ***P < 0.001.
Mentions: TRIM52 is a RING-type E3 ligase and it can mediate NS2A degradation via the ubiquitin-proteasome pathway. We thus speculated that the RING domain of TRIM52 also contributes to the antiviral activity of TRIM52. To validate this conjecture, we transfected 293T cells with plasmids encoding either the full-length TRIM52or RING domain deleted TRIM52 (TRIM52dR) or with empty vector 30 h prior to JEV infection at an MOI of 1. Progeny virus in culture supernatants was collected at 24 hpi, and viral titers were determined by plaque assay. The cells with full-length TRIM52 showed an obviously reduced viral production compared with the cells with TRIM52dR and empty vector control (Fig. 6a). By contrast, viral production in cells with TRIM52dR versus empty vector control did not vary (Fig. 6a). Moreover, JEV NS3 protein expression was significantly reduced in cells overexpressing the full-length TRIM52 compared with cells overexpressing TRIM52dR and vector control (Fig. 6b). No difference was observed between cells with TRIM52dR and empty vector (Fig. 6b). Overall, these results suggest that the RING domain of TRIM52 is required for TRIM52 to control JEV replication.

View Article: PubMed Central - PubMed

ABSTRACT

The members of tripartite-motif containing (TRIM) protein participate in various cellular processes and play an important role in host antiviral function. TRIM proteins exert their antiviral activity either directly by degrading viral proteins through their E3 ligase activity, or indirectly by promoting host innate immunity. This study demonstrated for the first time that TRIM52 is a novel antiviral TRIM protein against Japanese encephalitis virus (JEV) infection. Overexpression of TRIM52 restricted JEV replication in BHK-21 and 293T cells. In addition, JEV nonstructural protein 2A (NS2A) is a protein that interacts with TRIM52. Their interaction degraded NS2A in a proteasome-dependent manner via the E3 ligase activity of TRIM52. Thus, TRIM52 is a novel antiviral TRIM protein, and it exerted antiviral activity against JEV infection by targeting and degrading viral NS2A.

No MeSH data available.


Related in: MedlinePlus