Hepatitis A virus (HAV), a unique hepatotropic human picornavirus, is the causative agent of acute hepatitis A in humans. Some studies have shown that HAV antagonizes the innate immune response by disrupting interferon-beta (IFN-β) signaling by viral proteins. However, whether microRNAs (miRNAs), a class of non-coding RNAs, are involved in the antagonism of IFN-β induction upon HAV infection is still unclear. In this study, we investigated the effects and mechanisms by which HAV-induced miRNAs antagonize IFN-β signaling. A variety of analytical methods, including miRNA microarray, RT-qPCR, dual-luciferase reporter assay, and Western blotting, were performed using HAV-infected cells. The results indicated that HAV infection upregulates the expression of hsa-miR-146a-5p, which in turn partially suppresses the induction of IFN-β synthesis, thereby promoting viral replication. Mechanistically, TRAF6 (TNF receptor-associated factor 6), a key adaptor protein in the RIG-I/MDA5-mediated IFN-I signaling pathway, is targeted and degraded by hsa-miR-146a-5p. As TRAF6 is necessary for IFN-β induction, inhibition of this protein attenuates IFN-β signaling. Taken together, the results from this study indicated that HAV disrupts RIG-I/MDA5-mediated IFN-I signaling partially through the cleavage of the essential adaptor molecule TRAF6 via hsa-miR-146a-5p.
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Hepatitis A virus
Real-time reverse transcription PCR
Mitochondrial antiviral signaling protein
Tumor necrosis factor
Type I interferon
Toll-like receptor 3
Retinoic acid-inducible gene I
Oncorhynchus mykiss melanoma differentiation associated gene 5
Interferon-associated receptor 1/2
Janus kinase/signal transducer and activator of tran-ions
Nuclear factor-κB essential modulator
Tripartite motif containing 14
TANK-binding kinase 1
IΚB kinase ε
Interferon regulatory factor 3
C-C motif chemokine ligand 5
Kaposi’s sarcoma associated herpesvirus
Ubiquitin-specific peptidase 3
Vesicular stomatitis virus
Interleukin 1 receptor associated kinase 1/2
Herpes simplex virus
Interferon regulatory factor 1
Multiplicity of infection
Enzyme-linked immunosorbent assay
- TCID50 :
Tissue culture infectious dose
RNA integrity number
TIR domain-containing adaptor inducing IFN-β
Mitochondrial antiviral signaling protein
Myeloid differentiation primary response gene 88
TNF receptor associated factor 6
Receptor interacting serine/threonine kinase 3
Transmembrane protein 173
Ticehurst JR, Racaniello VR, Baroudy BM et al (1983) Molecular cloning and characterization of hepatitis A virus cDNA. Proc Natl Acad Sci USA 80:5885–5889
Najarian R, Caput D, Gee W et al (1985) Primary structure and gene organization of human hepatitis A virus. Proc Natl Acad Sci USA 82:2627–2631
Adams MJ, Lefkowitz EJ, King AMQ, Carstens EB (2014) Ratification vote on taxonomic proposals to the International Committee on Taxonomy of Viruses (2014). Arch Virol 159:2831–2841
Gauss-Müller V, Deinhardt F (1984) Effect of hepatitis A virus infection on cell metabolism in vitro. Proc Soc Exp Biol Med. 175:10–15
Vallbracht A, Hofmann L, Wurster KG, Flehmig B (1984) Persistent infection of human fibroblasts by hepatitis A virus. J Gen Virol. 65:609–615
Dotzauer A, Feinstone SM, Kaplan G (1994) Susceptibility of nonprimate cell lines to hepatitis A virus infection. J Virol. 68:6064–6068
Lanford RE, Zongdi F, Deborah C et al (2011) Acute hepatitis A virus infection is associated with a limited type I interferon response and persistence of intrahepatic viral RNA. Proc Natl Acad Sci USA 108:11223–11228
Vallbracht A, Gabriel P, Zahn J, Flehmig B (1985) Hepatitis A virus infection and the interferon system. J Infect Dis. 152:211–213
Kawai T, Akira S (2010) The role of pattern-recognition receptors in innate immunity: update on Toll-like receptors. Nat Immunol. 11:373–384
Satoshi U, Shizuo A (2006) Innate immune recognition of viral infection. Uirusu. 56:1–8
Takeuchi O, Akira S (2010) Recognition of viruses by innate immunity. Immunol Rev. 220:214–224
Paulmann D, Magulski T, Schwarz R et al (2008) Hepatitis A virus protein 2B suppresses beta interferon (IFN) gene transcription by interfering with IFN regulatory factor 3 activation. J Gen Virol. 89:1593–1604
Lin Q, Zongdi F, Daisuke Y et al (2011) Disruption of TLR3 signaling due to cleavage of TRIF by the hepatitis A virus protease-polymerase processing intermediate, 3CD. Plos Pathog. 7:e1002169
Yan Y, Yuqiong L, Lin Q et al (2007) Disruption of innate immunity due to mitochondrial targeting of a picornaviral protease precursor. Proc Natl Acad Sci USA. 104:7253–7258
Wang D, Fang L, Wei D et al (2014) Hepatitis A virus 3C protease cleaves NEMO to impair induction of beta interferon. J Virol. 88:10252–10258
Volker F, Dajana G, Iris B et al (2005) Hepatitis A virus suppresses RIG-I-mediated IRF-3 activation to block induction of beta interferon. J Virol. 79:10968–10977
Bhaskaran M, Mohan M (2014) MicroRNAs History, Biogenesis, and Their Evolving Role in Animal Development and Disease. Vet Pathol. 51:759–774
Houzet L, Jeang KT (2011) MicroRNAs and human retroviruses. BBA Gene Regul Me. 1809:686–693
Umbach JL, Cullen BR (2009) The role of RNAi and microRNAs in animal virus replication and antiviral immunity. Genes Dev. 23:1151–1164
Charles-Henri L, Patrice D, Khalil A et al (2005) A cellular microRNA mediates antiviral defense in human cells. Science. 308:557–560
Youngkyun K, Sanghyun L, Sungchul K et al (2012) Human cytomegalovirus clinical strain-specific microRNA miR-UL148D targets the human chemokine RANTES during infection. Plos Pathog. 8:e1002577
Liang D, Gao Y, Lin X et al (2011) A human herpesvirus miRNA attenuates interferon signaling and contributes to maintenance of viral latency by targeting IKKɛ. Cell Res. 21:793–806
Gao S, Li J, Song L, Wu J, Huang W (2017) Influenza A virus-induced downregulation of miR-26a contributes to reduced IFNα/β production. Virol Sin. 32:1–10
Changzhi X, Xiang H, Zirui Z et al (2014) Downregulation of microRNA miR-526a by enterovirus inhibits RIG-I-dependent innate immune response. J Virol. 88:11356–11368
Shi J, Duan Z, Sun J et al (2014) Identification and validation of a novel microRNA-like molecule derived from a cytoplasmic RNA virus antigenome by bioinformatics and experimental approaches. Virol J. 11:121–134
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402–408
Than LTL, Pei PC, Ng KP, Seow HF (2015) Detection of medically important candida species by absolute quantitation real-time polymerase chain reaction. Jundishapur J Microb 8:e14940
Saba R, Sorensen DL, Booth SA (2014) MicroRNA-146a: a dominant, negative regulator of the innate immune response. Front Immuno 5:578–588
Ho BC, Yu I, Lu LF et al (2014) Inhibition of miR-146a prevents enterovirus-induced death by restoring the production of type I interferon. Nat Commun 5:3344–3355
Zhang F, Sun X, Zhu Y, Qin W (2019) Downregulation of miR-146a inhibits influenza A virus replication by enhancing the type I interferon response in vitro and in vivo. Biomed Pharmacother 111:740–750
Hou J, Wang PL (2009) MicroRNA-146a feedback inhibits RIG-I-dependent Type I IFN production in macrophages by targeting TRAF6, IRAK1, and IRAK2. J Immunol 183:2150–2158
Wu S, He L, Li Y et al (2013) miR-146a facilitates replication of dengue virus by dampening interferon induction by targeting TRAF6. J Infect. 67:329–341
Walker CM, Feng Z, Lemon SM (2015) Reassessing immune control of hepatitis A virus. Curr Opin Virol. 11:7–13
Feng Z, Lemon SM (2019) Innate Immunity to Enteric Hepatitis Viruses. Cold Spring Harb Perspect Med 9:a033464
Bhaumik D, Scott GK, Schokrpur S et al (2008) Expression of microRNA-146 suppresses NF-kappaB activity with reduction of metastatic potential in breast cancer cells. Oncogene. 27:5643–5647
Hiroyasu K, Takuya Y, Kohsuke Y et al (2009) TRAF6 establishes innate immune responses by activating NF-kappaB and IRF7 upon sensing cytosolic viral RNA and DNA. Plos One. 4:e5674
This study was supported by the Chinese Academy of Medical Science (CAMS) Innovation Fund for Medical Sciences (2017-I2M-3-022), the National Natural Science Foundation of China (NSFC grant no. 31500724), the Applied Basic Research Key Project of Yunnan (CN) (202001AS070046), and the Fund for Reserve Talents of Young and Middle-Aged Academic and Technical Leaders of Yunnan Province (2019HB043).
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Mo, L., Zeng, Z., Deng, R. et al. Hepatitis A virus-induced hsa-miR-146a-5p attenuates IFN-β signaling by targeting adaptor protein TRAF6. Arch Virol 166, 789–799 (2021). https://doi.org/10.1007/s00705-021-04952-z