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TRIM30α Is a Negative-Feedback Regulator of the Intracellular DNA and DNA Virus-Triggered Response by Targeting STING.

Wang Y, Lian Q, Yang B, Yan S, Zhou H, He L, Lin G, Lian Z, Jiang Z, Sun B - PLoS Pathog. (2015)

Bottom Line: Homeostasis regulation of immune responses to cytosolic DNA is critical for limiting the risk of autoimmunity and survival of the host.Biochemical analyses showed that TRIM30α interacted with the stimulator of interferon genes (STING), which is a critical regulator of the DNA-sensing response.Overexpression of TRIM30α promoted the degradation of STING via K48-linked ubiquitination at Lys275 through a proteasome-dependent pathway.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China; School of Life Sciences, University of Science and Technology of China, Hefei, China.

ABSTRACT
Uncontrolled immune responses to intracellular DNA have been shown to induce autoimmune diseases. Homeostasis regulation of immune responses to cytosolic DNA is critical for limiting the risk of autoimmunity and survival of the host. Here, we report that the E3 ubiquitin ligase tripartite motif protein 30α (TRIM30α) was induced by herpes simplex virus type 1 (HSV-1) infection in dendritic cells (DCs). Knockdown or genetic ablation of TRIM30α augmented the type I IFNs and interleukin-6 response to intracellular DNA and DNA viruses. Trim30α-deficient mice were more resistant to infection by DNA viruses. Biochemical analyses showed that TRIM30α interacted with the stimulator of interferon genes (STING), which is a critical regulator of the DNA-sensing response. Overexpression of TRIM30α promoted the degradation of STING via K48-linked ubiquitination at Lys275 through a proteasome-dependent pathway. These findings indicate that E3 ligase TRIM30α is an important negative-feedback regulator of innate immune responses to DNA viruses by targeting STING.

No MeSH data available.


Related in: MedlinePlus

TRIM30α deficiency promotes immune signaling by cytoplasmic DNA and DNA virus infection in BMDCs.(A) Immunoblot analysis of TRIM30α expression in the splenocytes of wild type (WT) and Trim30α-/- (KO) mice. (B, C) ELISA of type I IFN and IL-6 in wild-type and Trim30α-/- BMDCs mock treated or stimulated for 16 h by transfection with poly(dA:dT) (1 μg/ml), ISD (1 μg/ml), c-di-GMP (8 μg/ml), genomic DNA (2 μg/ml) or poly(I:C) (5 μg/ml) (B); or with HSV-1 (MOI 10) or VACV (MOI 10) (C). (D) Immunoblot analysis of phosphorylated IRF3 (p-IRF3), p-p65 and TRIM30α in lysates of wild type and Trim30α-/- BMDCs stimulated for 1–5 h with poly(dA:dT) (1 μg/ml). Densitometry analysis to quantify ratio of p-IRF3 or p-p65 to β-actin is shown on the right. The data are representative of three independent experiments and are presented as mean ± SEM. *p < 0.05, **p < 0.01 and ***p < 0.001.
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ppat.1005012.g002: TRIM30α deficiency promotes immune signaling by cytoplasmic DNA and DNA virus infection in BMDCs.(A) Immunoblot analysis of TRIM30α expression in the splenocytes of wild type (WT) and Trim30α-/- (KO) mice. (B, C) ELISA of type I IFN and IL-6 in wild-type and Trim30α-/- BMDCs mock treated or stimulated for 16 h by transfection with poly(dA:dT) (1 μg/ml), ISD (1 μg/ml), c-di-GMP (8 μg/ml), genomic DNA (2 μg/ml) or poly(I:C) (5 μg/ml) (B); or with HSV-1 (MOI 10) or VACV (MOI 10) (C). (D) Immunoblot analysis of phosphorylated IRF3 (p-IRF3), p-p65 and TRIM30α in lysates of wild type and Trim30α-/- BMDCs stimulated for 1–5 h with poly(dA:dT) (1 μg/ml). Densitometry analysis to quantify ratio of p-IRF3 or p-p65 to β-actin is shown on the right. The data are representative of three independent experiments and are presented as mean ± SEM. *p < 0.05, **p < 0.01 and ***p < 0.001.

Mentions: To further demonstrate the function of TRIM30α in immune response to DNA-mediated signaling, we generated Trim30α-deficient mice, in which the second exon was knocked out by homologous recombination (S2 Fig). Immunoblot analysis confirmed that TRIM30α expression was completely absent in Trim30α-deficient mice (Fig 2A). As shown in Fig 2B, TRIM30α deficiency resulted in much higher expression of type I IFN and IL-6 in response to multiple DNA ligands compared with wild-type DCs, similar to the results observed in D2SC cells. In contrast, TRIM30α deficiency did not affect poly(I:C) signaling (Fig 2B). Consistently with previous results, TRIM30α deficiency dramatically increased IFN-β and IL-6 expression induced by HSV-1 and VACV infection at protein level (Fig 2C). Beside the role in DNA sensing pathway, we also determine the function of TRIM30α against RNA viruses. As shown in S3 Fig, TRIM30α knockdown or deficiency dramatically potentiated IFN-β and IL-6 production by infection with VSV in D2SC cells and BMDCs (S3 Fig). The results suggest that TRIM30α negatively regulate immune response both to DNA and RNA viruses.


TRIM30α Is a Negative-Feedback Regulator of the Intracellular DNA and DNA Virus-Triggered Response by Targeting STING.

Wang Y, Lian Q, Yang B, Yan S, Zhou H, He L, Lin G, Lian Z, Jiang Z, Sun B - PLoS Pathog. (2015)

TRIM30α deficiency promotes immune signaling by cytoplasmic DNA and DNA virus infection in BMDCs.(A) Immunoblot analysis of TRIM30α expression in the splenocytes of wild type (WT) and Trim30α-/- (KO) mice. (B, C) ELISA of type I IFN and IL-6 in wild-type and Trim30α-/- BMDCs mock treated or stimulated for 16 h by transfection with poly(dA:dT) (1 μg/ml), ISD (1 μg/ml), c-di-GMP (8 μg/ml), genomic DNA (2 μg/ml) or poly(I:C) (5 μg/ml) (B); or with HSV-1 (MOI 10) or VACV (MOI 10) (C). (D) Immunoblot analysis of phosphorylated IRF3 (p-IRF3), p-p65 and TRIM30α in lysates of wild type and Trim30α-/- BMDCs stimulated for 1–5 h with poly(dA:dT) (1 μg/ml). Densitometry analysis to quantify ratio of p-IRF3 or p-p65 to β-actin is shown on the right. The data are representative of three independent experiments and are presented as mean ± SEM. *p < 0.05, **p < 0.01 and ***p < 0.001.
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ppat.1005012.g002: TRIM30α deficiency promotes immune signaling by cytoplasmic DNA and DNA virus infection in BMDCs.(A) Immunoblot analysis of TRIM30α expression in the splenocytes of wild type (WT) and Trim30α-/- (KO) mice. (B, C) ELISA of type I IFN and IL-6 in wild-type and Trim30α-/- BMDCs mock treated or stimulated for 16 h by transfection with poly(dA:dT) (1 μg/ml), ISD (1 μg/ml), c-di-GMP (8 μg/ml), genomic DNA (2 μg/ml) or poly(I:C) (5 μg/ml) (B); or with HSV-1 (MOI 10) or VACV (MOI 10) (C). (D) Immunoblot analysis of phosphorylated IRF3 (p-IRF3), p-p65 and TRIM30α in lysates of wild type and Trim30α-/- BMDCs stimulated for 1–5 h with poly(dA:dT) (1 μg/ml). Densitometry analysis to quantify ratio of p-IRF3 or p-p65 to β-actin is shown on the right. The data are representative of three independent experiments and are presented as mean ± SEM. *p < 0.05, **p < 0.01 and ***p < 0.001.
Mentions: To further demonstrate the function of TRIM30α in immune response to DNA-mediated signaling, we generated Trim30α-deficient mice, in which the second exon was knocked out by homologous recombination (S2 Fig). Immunoblot analysis confirmed that TRIM30α expression was completely absent in Trim30α-deficient mice (Fig 2A). As shown in Fig 2B, TRIM30α deficiency resulted in much higher expression of type I IFN and IL-6 in response to multiple DNA ligands compared with wild-type DCs, similar to the results observed in D2SC cells. In contrast, TRIM30α deficiency did not affect poly(I:C) signaling (Fig 2B). Consistently with previous results, TRIM30α deficiency dramatically increased IFN-β and IL-6 expression induced by HSV-1 and VACV infection at protein level (Fig 2C). Beside the role in DNA sensing pathway, we also determine the function of TRIM30α against RNA viruses. As shown in S3 Fig, TRIM30α knockdown or deficiency dramatically potentiated IFN-β and IL-6 production by infection with VSV in D2SC cells and BMDCs (S3 Fig). The results suggest that TRIM30α negatively regulate immune response both to DNA and RNA viruses.

Bottom Line: Homeostasis regulation of immune responses to cytosolic DNA is critical for limiting the risk of autoimmunity and survival of the host.Biochemical analyses showed that TRIM30α interacted with the stimulator of interferon genes (STING), which is a critical regulator of the DNA-sensing response.Overexpression of TRIM30α promoted the degradation of STING via K48-linked ubiquitination at Lys275 through a proteasome-dependent pathway.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China; School of Life Sciences, University of Science and Technology of China, Hefei, China.

ABSTRACT
Uncontrolled immune responses to intracellular DNA have been shown to induce autoimmune diseases. Homeostasis regulation of immune responses to cytosolic DNA is critical for limiting the risk of autoimmunity and survival of the host. Here, we report that the E3 ubiquitin ligase tripartite motif protein 30α (TRIM30α) was induced by herpes simplex virus type 1 (HSV-1) infection in dendritic cells (DCs). Knockdown or genetic ablation of TRIM30α augmented the type I IFNs and interleukin-6 response to intracellular DNA and DNA viruses. Trim30α-deficient mice were more resistant to infection by DNA viruses. Biochemical analyses showed that TRIM30α interacted with the stimulator of interferon genes (STING), which is a critical regulator of the DNA-sensing response. Overexpression of TRIM30α promoted the degradation of STING via K48-linked ubiquitination at Lys275 through a proteasome-dependent pathway. These findings indicate that E3 ligase TRIM30α is an important negative-feedback regulator of innate immune responses to DNA viruses by targeting STING.

No MeSH data available.


Related in: MedlinePlus