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Neddylation is required for herpes simplex virus type I (HSV-1)-induced early phase interferon-beta production

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ABSTRACT

Type I interferons such as interferon-beta (IFN-β) play essential roles in the host innate immune response to herpes simplex virus type I (HSV-1) infection. The transcription of type I interferon genes is controlled by nuclear factor-κB (NF-κB) and interferon regulatory factor (IRF) family members including IRF3. NF-κB activation depends on the phosphorylation of inhibitor of κB (IκB), which triggers its ubiqitination and degradation. It has been reported that neddylation inhibition by a pharmacological agent MLN4924 potently suppresses lipopolysaccharide (LPS)-induced proinflammatory cytokine production with the accumulation of phosphorylated IκBα. However, the role of neddylation in type I interferon expression remains unknown. Here, we report that neddylation inhibition with MLN4924 or upon UBA3 deficiency led to accumulation of phosphorylated IκBα, impaired IκBα degradation, and impaired NF-κB nuclear translocation in the early phase of HSV-1 infection even though phosphorylation and nuclear translocation of IRF3 were not affected. The blockade of NF-κB nuclear translocation by neddylation inhibition becomes less efficient at the later time points of HSV-1 infection. Consequently, HSV-1-induced early phase IFN-β production significantly decreased upon MLN4924 treatment and UBA3 deficiency. NF-κB inhibitor JSH-23 mimicked the effects of neddylation inhibition in the early phase of HSV-1 infection. Moreover, the effects of neddylation inhibition on HSV-1-induced early phase IFN-β production diminished in the presence of NF-κB inhibitor JSH-23. Thus, neddylation contributes to HSV-1-induced early phase IFN-β production through, at least partially, promoting NF-κB activation.

No MeSH data available.


Neddylation inhibition dampens HSV-1-induced early phase IFN-β production. BMMs were pretreated with 0.1 μM MLN4924 or DMSO of equal volume for 30 min (a,c) or BMMs were cultured from UBA3F/F and UBA3ΔMye mice (b,d). Then BMMs were infected with HSV-1 for the indicated periods of time. Relative IFN-β mRNA expression was measured by qRT-PCR (a,b) and IFN-β concentration in the supernatants was measured with ELISA (c,d). **p < 0.01.
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fig6: Neddylation inhibition dampens HSV-1-induced early phase IFN-β production. BMMs were pretreated with 0.1 μM MLN4924 or DMSO of equal volume for 30 min (a,c) or BMMs were cultured from UBA3F/F and UBA3ΔMye mice (b,d). Then BMMs were infected with HSV-1 for the indicated periods of time. Relative IFN-β mRNA expression was measured by qRT-PCR (a,b) and IFN-β concentration in the supernatants was measured with ELISA (c,d). **p < 0.01.

Mentions: Since neddylation inhibition upon MLN4924 pretreatment or UBA3 deficiency leads to impaired NF-κB activation in the early phase of HSV-1 infection, we then explored whether neddylation inhibition affected IFN-β expression with quantitative real-time reverse-transcriptase PCR. As expected, the level of IFN-β mRNA was significantly upregulated 4 h after HSV-1 infection, which dropped about 50% 24 h after HSV-1 infection (Figure 6a,b). A measure of 0.1 μM MLN4924 pretreatment for 30 min led to about 50% reduction of IFN-β mRNA 4 h after HSV-1 infection (Figure 6a). The inhibitory effect diminished 24 h after HSV-1 infection (Figure 6a). UBA3 deficiency exhibited similar effects on HSV-1-induced upregulation of IFN-β mRNA (Figure 6b).


Neddylation is required for herpes simplex virus type I (HSV-1)-induced early phase interferon-beta production
Neddylation inhibition dampens HSV-1-induced early phase IFN-β production. BMMs were pretreated with 0.1 μM MLN4924 or DMSO of equal volume for 30 min (a,c) or BMMs were cultured from UBA3F/F and UBA3ΔMye mice (b,d). Then BMMs were infected with HSV-1 for the indicated periods of time. Relative IFN-β mRNA expression was measured by qRT-PCR (a,b) and IFN-β concentration in the supernatants was measured with ELISA (c,d). **p < 0.01.
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Related In: Results  -  Collection

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fig6: Neddylation inhibition dampens HSV-1-induced early phase IFN-β production. BMMs were pretreated with 0.1 μM MLN4924 or DMSO of equal volume for 30 min (a,c) or BMMs were cultured from UBA3F/F and UBA3ΔMye mice (b,d). Then BMMs were infected with HSV-1 for the indicated periods of time. Relative IFN-β mRNA expression was measured by qRT-PCR (a,b) and IFN-β concentration in the supernatants was measured with ELISA (c,d). **p < 0.01.
Mentions: Since neddylation inhibition upon MLN4924 pretreatment or UBA3 deficiency leads to impaired NF-κB activation in the early phase of HSV-1 infection, we then explored whether neddylation inhibition affected IFN-β expression with quantitative real-time reverse-transcriptase PCR. As expected, the level of IFN-β mRNA was significantly upregulated 4 h after HSV-1 infection, which dropped about 50% 24 h after HSV-1 infection (Figure 6a,b). A measure of 0.1 μM MLN4924 pretreatment for 30 min led to about 50% reduction of IFN-β mRNA 4 h after HSV-1 infection (Figure 6a). The inhibitory effect diminished 24 h after HSV-1 infection (Figure 6a). UBA3 deficiency exhibited similar effects on HSV-1-induced upregulation of IFN-β mRNA (Figure 6b).

View Article: PubMed Central - PubMed

ABSTRACT

Type I interferons such as interferon-beta (IFN-&beta;) play essential roles in the host innate immune response to herpes simplex virus type I (HSV-1) infection. The transcription of type I interferon genes is controlled by nuclear factor-&kappa;B (NF-&kappa;B) and interferon regulatory factor (IRF) family members including IRF3. NF-&kappa;B activation depends on the phosphorylation of inhibitor of &kappa;B (I&kappa;B), which triggers its ubiqitination and degradation. It has been reported that neddylation inhibition by a pharmacological agent MLN4924 potently suppresses lipopolysaccharide (LPS)-induced proinflammatory cytokine production with the accumulation of phosphorylated I&kappa;B&alpha;. However, the role of neddylation in type I interferon expression remains unknown. Here, we report that neddylation inhibition with MLN4924 or upon UBA3 deficiency led to accumulation of phosphorylated I&kappa;B&alpha;, impaired I&kappa;B&alpha; degradation, and impaired NF-&kappa;B nuclear translocation in the early phase of HSV-1 infection even though phosphorylation and nuclear translocation of IRF3 were not affected. The blockade of NF-&kappa;B nuclear translocation by neddylation inhibition becomes less efficient at the later time points of HSV-1 infection. Consequently, HSV-1-induced early phase IFN-&beta; production significantly decreased upon MLN4924 treatment and UBA3 deficiency. NF-&kappa;B inhibitor JSH-23 mimicked the effects of neddylation inhibition in the early phase of HSV-1 infection. Moreover, the effects of neddylation inhibition on HSV-1-induced early phase IFN-&beta; production diminished in the presence of NF-&kappa;B inhibitor JSH-23. Thus, neddylation contributes to HSV-1-induced early phase IFN-&beta; production through, at least partially, promoting NF-&kappa;B activation.

No MeSH data available.