Influenza A Virus-induced expression of ISG20 inhibits viral replication by interacting with nucleoprotein

Qu, Hongren; Li, Jing; Yang, Limin; Sun, Lei; Liu, Wenjun; He, Hongxuan

doi:10.1007/s11262-016-1366-2

Cited by 44 publications

(46 citation statements)

References 43 publications

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“…RuvB-like protein 2 disrupts vRNP assembly by interfering with NP oligomerization (34). Interferon-stimulated gene 20 interacts with NP and inhibits viral protein expression (35). Similarly, our study showed that TRIM41 modulates IAV infection by interaction with NP, which allows NP ubiquitination and subsequent protein degradation.…”

Section: Discussionsupporting

confidence: 54%

TRIM41-Mediated Ubiquitination of Nucleoprotein Limits Influenza A Virus Infection

Patil

Zhao

Song

et al. 2018

J Virol

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Influenza A virus (IAV) is a highly transmissible respiratory pathogen and a major cause of morbidity and mortality around the world. Nucleoprotein (NP) is an abundant IAV protein essential for multiple steps of the viral life cycle. Our recent proteomic study of the IAV-host interaction network found that TRIM41 (tripartite motif-containing 41), a ubiquitin E3 ligase, interacted with NP. However, the role of TRIM41 in IAV infection is unknown. Here, we report that TRIM41 interacts with NP through its SPRY domain. Furthermore, TRIM41 is constitutively expressed in lung epithelial cells, and overexpression of TRIM41 inhibits IAV infection. Conversely, RNA interference (RNAi) and knockout of TRIM41 increase host susceptibility to IAV infection. As a ubiquitin E3 ligase, TRIM41 ubiquitinates NP and in cells. The TRIM41 mutant lacking E3 ligase activity fails to inhibit IAV infection, suggesting that the E3 ligase activity is indispensable for TRIM41 antiviral function. Mechanistic analysis further revealed that the polyubiquitination leads to NP protein degradation and viral inhibition. Taking these observations together, TRIM41 is a constitutively expressed intrinsic IAV restriction factor that targets NP for ubiquitination and protein degradation. Influenza control strategies rely on annual immunization and require frequent updates of the vaccine, which is not always a foolproof process. Furthermore, the current antivirals are also losing effectiveness as new viral strains are often refractory to conventional treatments. Thus, there is an urgent need to find new antiviral mechanisms and develop therapeutic drugs based on these mechanisms. Targeting the virus-host interface is an emerging new strategy because host factors controlling viral replication activity will be ideal candidates, and cellular proteins are less likely to mutate under drug-mediated selective pressure. Here, we show that the ubiquitin E3 ligase TRIM41 is an intrinsic host restriction factor to IAV. TRIM41 directly binds the viral nucleoprotein and targets it for ubiquitination and proteasomal degradation, thereby limiting viral infection. Exploitation of this natural defense pathway may open new avenues to develop antiviral drugs targeting the influenza virus.

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Section: Discussionsupporting

confidence: 54%

TRIM41-Mediated Ubiquitination of Nucleoprotein Limits Influenza A Virus Infection

Patil

Zhao

Song

et al. 2018

J Virol

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“…However, RNA degradation might not be the only mechanism of ISG20's antiviral action. For instance, Zhou et al found that ISG20 did not promote the degradation of a replication-defective HCV RNA genome in Huh7.5 cells [17]; and Qu et al observed that ISG20 inhibits Influenza A Virus replication through interaction with viral nucleoproteins [37]. Furthermore, not all the RNA viruses are susceptible to ISG20, as severe acute respiratory syndrome coronavirus (SARS-CoV) has shown resistance to ISG20 in cell cultures [17], indicating that there is viral specific factor(s) to determine the antiviral specificity of ISG20.…”

Section: Discussionmentioning

confidence: 99%

Interferon-inducible ribonuclease ISG20 inhibits hepatitis B virus replication through directly binding to the epsilon stem-loop structure of viral RNA

et al. 2017

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Hepatitis B virus (HBV) replicates its DNA genome through reverse transcription of a viral RNA pregenome. We report herein that the interferon (IFN) stimulated exoribonuclease gene of 20 KD (ISG20) inhibits HBV replication through degradation of HBV RNA. ISG20 expression was observed at basal level and was highly upregulated upon IFN treatment in hepatocytes, and knock down of ISG20 resulted in elevation of HBV replication and attenuation of IFN-mediated antiviral effect. The sequence element conferring the susceptibility of HBV RNA to ISG20-mediated RNA degradation was mapped at the HBV RNA terminal redundant region containing epsilon (ε) stem-loop. Furthermore, ISG20-induced HBV RNA degradation relies on its ribonuclease activity, as the enzymatic inactive form ISG20D94G was unable to promote HBV RNA decay. Interestingly, ISG20D94G retained antiviral activity against HBV DNA replication by preventing pgRNA encapsidation, resulting from a consequence of ISG20-ε interaction. This interaction was further characterized by in vitro electrophoretic mobility shift assay (EMSA) and ISG20 was able to bind HBV ε directly in absence of any other cellular proteins, indicating a direct ε RNA binding capability of ISG20; however, cofactor(s) may be required for ISG20 to efficiently degrade ε. In addition, the lower stem portion of ε is the major ISG20 binding site, and the removal of 4 base pairs from the bottom portion of ε abrogated the sensitivity of HBV RNA to ISG20, suggesting that the specificity of ISG20-ε interaction relies on both RNA structure and sequence. Furthermore, the C-terminal Exonuclease III (ExoIII) domain of ISG20 was determined to be responsible for interacting with ε, as the deletion of ExoIII abolished in vitro ISG20-ε binding and intracellular HBV RNA degradation. Taken together, our study sheds light on the underlying mechanisms of IFN-mediated HBV inhibition and the antiviral mechanism of ISG20 in general.

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“…Moreover, ISG20, a member of the 3=-to-5= exonuclease superfamily, displays effective antiviral activity against several RNA viruses, including IAV, vesicular stomatitis virus (VSV), hepatitis C virus (HCV), and encephalomyocarditis virus (EMCV), mostly relying on its RNase activity (12)(13)(14)(15)(16). In addition, ISG20 inhibits IAV replication by interacting with viral NP protein and inhibiting viral polymerase activity (17).…”

Section: Importancementioning

confidence: 99%

Lnc-ISG20 Inhibits Influenza A Virus Replication by Enhancing ISG20 Expression

Chai

Shangguan

et al. 2018

J Virol

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Long noncoding RNAs (lncRNAs) are involved in many aspects of cellular processes, including the antiviral immune response. To identify influenza A virus (IAV)-related lncRNAs, we performed RNA deep sequencing to compare the profiles of lncRNAs in A549 and HEK293T cells with or without IAV infection. We identified an IAV-upregulated lncRNA named lnc-ISG20 because it shares most of its sequence with ISG20. We found that lnc-ISG20 is an interferon-stimulated gene similar to ISG20. Overexpression of lnc-ISG20 inhibited IAV replication, while lnc-ISG20 knockdown favored viral replication, suggesting that lnc-ISG20 is inhibitory to IAV replication. Further study indicated that overexpression of lnc-ISG20 enhances ISG20 protein levels, while knockdown of lnc-ISG20 reduces ISG20 protein levels in A549 cells induced with poly(I·C) and Sendai virus. We demonstrated that lnc-ISG20 inhibits IAV replication in an ISG20-dependent manner. As lnc-ISG20 did not affect the mRNA level of ISG20, we postulated that lnc-ISG20 may function as endogenous RNA competing with ISG20 to enhance its translation. Indeed, we identified that microRNA 326 (miR-326) is a mutual microRNA for both ISG20 and lnc-ISG20 that targets the 3' untranslated region of ISG20 mRNA to inhibit its translation. We confirmed that lnc-ISG20 can bind miR-326, which in turn decreased the amount of miR-326 bound to ISG20 mRNA. In conclusion, we identified that the IAV-upregulated lnc-ISG20 is a novel interferon-stimulated gene that elicits its inhibitory effect on IAV replication by enhancing ISG20 expression. We demonstrated that lnc-ISG20 functions as a competitive endogenous RNA to bind miR-326 to reduce its inhibition of ISG20 translation. Our results revealed the mechanism by which lnc-ISG20 inhibits IAV replication. The replication of influenza A virus is regulated by host factors. However, the mechanisms by which lncRNAs regulate IAV infection are not well understood. We identified that lnc-ISG20 is upregulated during IAV infection and is also an interferon-stimulated gene. We demonstrated that lnc-ISG20 can enhance ISG20 expression, which in turn inhibits IAV replication. Our studies indicate that lnc-ISG20 functions as a competing endogenous RNA that binds miR-326 and reduces its inhibitory effect on ISG20. Taken together, our findings reveal the mechanistic details of lnc-ISG20 negatively regulating IAV replication. These findings indicate that lnc-ISG20 plays an important role during the host antiviral immune response.

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Influenza A Virus-induced expression of ISG20 inhibits viral replication by interacting with nucleoprotein

Cited by 44 publications

References 43 publications

TRIM41-Mediated Ubiquitination of Nucleoprotein Limits Influenza A Virus Infection

TRIM41-Mediated Ubiquitination of Nucleoprotein Limits Influenza A Virus Infection

Interferon-inducible ribonuclease ISG20 inhibits hepatitis B virus replication through directly binding to the epsilon stem-loop structure of viral RNA

Lnc-ISG20 Inhibits Influenza A Virus Replication by Enhancing ISG20 Expression

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