Skip to main content

Advertisement

SpringerLink
Log in

MicroRNAs miR-18a and miR-452 regulate the replication of enterovirus 71 by targeting the gene encoding VP3

  • Original Paper
  • Published:
Virus Genes Aims and scope Submit manuscript

Abstract

MicroRNAs (miRNAs) are crucial in the process of host–pathogen interaction. In this study, we established a screening system for miRNAs of target genes to detect the effect of miRNAs on Enterovirus 71 (EV71) replication in rhabdomyosarcoma (RD) cells. A 3’-untranslated region (UTR) dual-luciferase assay was performed to confirm putative miRNA targets in EV71 genome. Firstly, 13 fragments of EV71 genome were inserted into the vector pMIR, and luciferase activities were analyzed to identify the putative miRNAs of target genes. The expression of the reporter protein was significantly downregulated in cells transfected with the vector containing gene VP3. Then we screened for miRNAs that might target to VP3 through online analysis software. In addition, Western blot, real-time PCR, virus titration, and morphological changes were considered to examine the effects of miRNAs on virus replication. The results suggested that miR-18a and miR-452 repress the reproduction of EV71 virus by binding to VP3. Moreover, EV71 infection also affected the expression of endogenous miR-18a and miR-452. In addition, no significant cytotoxic effects were observed. The results from this study suggest that the intracellular miRNAs may play vital roles in the host–virus interaction.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Data availability

All data presented in this study are available from the corresponding author upon reasonable request.

References

  1. Ayukekbong JA, Bergström T (2014) Polio will go, acute flaccid paralysis will stay. Lancet 383:2209–2210

    Article  Google Scholar 

  2. Chen KT, Chang HL, Wang ST, Cheng YT, Yang JY (2007) Epidemiologic features of hand–foot–mouth disease and herpangina caused by enterovirus 71 in Taiwan, 1998–2005. Pediatrics 120:244–252

    Article  Google Scholar 

  3. Ooi MH, Wong SC, Lewthwaite P, Cardosa MJ, Solomon T (2010) Clinical features, diagnosis, and management of enterovirus 71. Lancet Neurol 9:1097–1105

    Article  Google Scholar 

  4. Wong KT, Munisamy B, Ong KC, Kojima H, Noriyo N, Chua KB, Ong BB, Nagashima K (2008) The distribution of inflammation and virus in human enterovirus 71 encephalomyelitis suggests possible viral spread by neural pathways. J Neuropathol Exp Neurol 67:162–169

    Article  Google Scholar 

  5. Cox JA, Hiscox JA, Solomon T, Ooi MH, Ng LFP (2017) Immunopathogenesis and virus-host interactions of enterovirus 71 in patients with hand, foot and mouth disease. Front Microbiol 8:2249

    Article  Google Scholar 

  6. Lee JA, Yoon YS, Hyeon JY, Yoo JS, Lee SW, Lee JW, Lee SW (2016) Sequence analysis of the first C2 subgenogroup strain of enterovirus 71 isolated in Korea. J Clin Virol 85:13–16

    Article  CAS  Google Scholar 

  7. Xing W, Liao Q, Viboud C, Zhang J, Sun J, Wu JT et al (2014) Hand, foot, and mouth disease in China, 2008–12: an epidemiological study. Lancet Infect Dis 14:308–318

    Article  Google Scholar 

  8. Chen M, Ju Y, Chen M, Xie Z, Zhou K, Tan Y, Mo J (2017) Epidemiological and genetic characteristics of EV71 in hand, foot, and mouth disease in Guangxi, southern China, from 2010 to 2015. PLoS ONE 12:e0188640

    Article  Google Scholar 

  9. Yates LA, Norbury CJ, Gilbert RJ (2013) The long and short of microRNA. Cell 153:516–519

    Article  CAS  Google Scholar 

  10. Shukla GC, Singh J, Barik S (2011) MicroRNAs: processing, maturation, target recognition and regulatory functions. Mol Cell Pharmacol 3:83–92

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Ebert M, Sharp PA (2012) Roles for microRNAs in conferring robustness to biological processes. Cell 149:515–524

    Article  CAS  Google Scholar 

  12. Kim J, Yao F, Xiao Z, Sun Y, Ma L (2018) MicroRNAs and metastasis: small RNAs play big roles. Cancer Metastasis Rev 37:5–15

    Article  CAS  Google Scholar 

  13. Modai S, Farberov L, Herzig E, Isakov O, Hizi A, Shomron N (2019) HIV-1 infection increases microRNAs that inhibit Dicer1, HRB and HIV-EP2, thereby reducing viral replication. PLoS ONE 14:e0211111

    Article  CAS  Google Scholar 

  14. Wang Y, Li Y (2018) miR-146 promotes HBV replication and expression by targeting ZEB2. Biomed Pharmacother 99:576–582

    Article  CAS  Google Scholar 

  15. Wang R, Zhang YY, Lu JS, Xia BH, Yang ZX, Zhu XD, Zhou XW, Huang PT (2017) The highly pathogenic H5N1 influenza A virus down-regulated several cellular MicroRNAs which target viral genome. J Cell Mol Med 21:3076–3086

    Article  CAS  Google Scholar 

  16. Wang RYL, Weng KF, Huang YC, Chen CJ (2016) Elevated expression of circulating miR876-5p is a specific response to severe EV71 infections. Sci Rep 6:24149

    Article  CAS  Google Scholar 

  17. Sun Y, Feng L, Li J, Xu H, Mei X, Feng L, Sun H, Gao J, Zhang X (2019) miR-545 promoted enterovirus 71 replication via directly targeting phosphatase and tensin homolog and tumor necrosis factor receptor-associated factor 6. J Cell Physiol. https://doi.org/10.1002/jcp.28222

    Article  PubMed  PubMed Central  Google Scholar 

  18. Zhao Q, Xiong Y, Xu J, Chen S, Li P, Huang Y, Wang Y, Chen WX, Wang B (2018) Host microRNA hsa-miR-494-3p promotes EV71 replication by directly targeting PTEN. Front Cell Infect Microbiol 8:278

    Article  Google Scholar 

  19. Zhang L, Chen X, Shi Y, Zhou B, Du C, Liu Y (2014) miR-27a suppresses EV71 replication by directly targeting EGFR. Virus Genes 49:373–382

    Article  CAS  Google Scholar 

  20. Zheng Z, Ke X, Wang M, He S, Li Q, Zheng C, Zhang Z, Liu Y, Wang H (2013) Human microRNA hsa-miR-296-5p suppresses enterovirus 71 replication by targeting the viral genome. J Virol 87:5645–5656

    Article  CAS  Google Scholar 

  21. Wen BP, Dai HJ, Yang YH, Zhuang Y, Sheng R (2013) MicroRNA-23b inhibits enterovirus 71 replication through downregulation of EV71 VP1 protein. Intervirology 56:195–200

    Article  CAS  Google Scholar 

  22. Wang B, Zhang H, Zhu M, Luo Z, Peng Y (2012) MEK1-ERKs signal cascade is required for the replication of enterovirus 71 (EV71). Antivir Res 93:110–117

    Article  CAS  Google Scholar 

  23. Hsu PW, Lin LZ, Hsu SD, Hsu JB, Huang HD (2007) ViTa: prediction of host microRNAs targets on viruses. Nucleic Acids Res 35:D381–D385

    Article  CAS  Google Scholar 

  24. Ventsislav R, Vesselin B, Ivan NM, Martin T (2005) MicroInspector: a web tool for detection of miRNA binding sites in an RNA sequence. Nucleic Acids Res 33:W696–W700

    Article  Google Scholar 

  25. Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 3:1101–1108

    Article  CAS  Google Scholar 

  26. Lin JY, Chen TC, Weng KF, Chang SC, Chen LL, Shih SR (2009) Viral and host proteins involved in picornavirus life cycle. J Biomed Sci 16:103

    Article  Google Scholar 

  27. Yi EJ, Shin YJ, Kim JH, Kim TG, Chang SY (2017) Enterovirus 71 infection and vaccines. Clin Exp Vaccine Res 6:4–14

    Article  CAS  Google Scholar 

  28. Gao F, Wang YP, Mao QY, Yao X, Liu S, Li FX et al (2012) Enterovirus 71 viral capsid protein linear epitopes: identification and characterization. J Virol 9:26

    Article  CAS  Google Scholar 

  29. Lei X, Liu X, Ma Y, Sun Z, Yang Y, Jin Q, He B, Wang J (2010) The 3C protein of enterovirus 71 inhibits retinoid acid-inducible gene I-mediated interferon regulatory factor 3 activation and type I interferon responses. J Virol 84:8051–8061

    Article  CAS  Google Scholar 

  30. Lecellier CH, Dunoyer P, Arar K, Che JL, Eyquem S, Himber C, Saib A, Voinnet O (2005) A cellular microRNA mediates antiviral defense in human cells. Science 308:557–560

    Article  CAS  Google Scholar 

  31. Otsuka M, Jing Q, Georgel P, New L, Chen J, Mols J, Kang YJ, Jiang Z, Du X, Cook R, Das SC, Pattnaik AK, Beutler B, Han J (2007) Hypersusceptibility to vesicular stomatitis virus infection in Dicer1-deficient mice is due to impaired miR24 and miR93 expression. Immunity 27:123–134

    Article  CAS  Google Scholar 

  32. Coppola N, Potenza N, Pisaturo M, Mosca N, Tonziello G, Signoriello G, Messina V, Sagnelli C, Russo A, Sagnelli E (2013) Liver microRNA hsa-miR-125a-5p in HBV chronic infection: correlation with HBV replication and disease progression. PLoS ONE 8:e65336

    Article  CAS  Google Scholar 

  33. Chen YN, Shen A, Rider PJ, Yu Y, Wu KL, Mu YX, Hao Q, Liu YL, Gong H, Zhu Y, Liu FY, Wu JG (2011) A liver-specific microRNA binds to a highly conserved RNA sequence of hepatitis B virus and negatively regulates viral gene expression and replication. FASEB J 25:4511–4521

    Article  CAS  Google Scholar 

  34. Chang J, Guo JT, Jiang D, Guo H, Taylor JM, Block TM (2008) Liver-specific microRNA miR-122 enhances the replication of hepatitis C virus in nonhepatic cells. J Virol 82:8215–8223

    Article  CAS  Google Scholar 

  35. Song L, Liu H, Gao S, Jiang W, Huang W (2010) Cellular microRNAs inhibit replication of the H1N1 influenza A virus in infected cells. J Virol 84:8849–8860

    Article  CAS  Google Scholar 

Download references

Funding

This study was supported by grant from Operating Project of Public Health Emergency Response Mechanism of National Institute for Nutrition and Health, C CDC (131031107000160002).

Author information

Authors and Affiliations

  1. National Institute for Nutrition and Health, Chinese Center for Disease Control and Prevention, Beijing, 100050, China

    Zhuo Yang, Qin Zhuo, Wen Qin, Jingbo Wang & Liyuan Wang

  2. Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China

    Po Tien

Authors
  1. Zhuo Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qin Zhuo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wen Qin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jingbo Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Liyuan Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Po Tien
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

ZY designed the study. ZY, WQ, JW, and LW performed the experiments. ZY analyzed and interpreted data. ZY wrote, reviewed, and revisioned the manuscript. QZ and PT provided administrative, technical, and material support.

Corresponding author

Correspondence to Zhuo Yang.

Ethics declarations

Conflict of interest

The authors declare no competing financial interests.

Ethical approval

This article does not contain any studies on animals performed by any of the authors.

Additional information

Edited by Joachim Jakob Bugert.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Fig.S1

Analysis of the specificity and cytotoxicity of miR-18a and miR-452. a Viability of RD cells (as an indicator of cytotoxicity) was determined by the CCK-8 assay after transfection with miRNA mimics or inhibitor. Absorbance values of the cells were measured at 450 nm. b The levels of IFN-α and IFN-β in the cells were determined by measuring the absorbance values at 450 nm (JPG 259 kb)

Supplementary file2 (DOCX 16 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yang, Z., Zhuo, Q., Qin, W. et al. MicroRNAs miR-18a and miR-452 regulate the replication of enterovirus 71 by targeting the gene encoding VP3. Virus Genes 57, 318–326 (2021). https://doi.org/10.1007/s11262-021-01842-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11262-021-01842-z

Keywords

Search

Navigation