1887

Abstract

Bovine herpesvirus 5 (BoHV-5) is a pathogen of cattle responsible for fatal meningoencephalitis. Like alpha herpesvirus subfamily members, BoHV-5 also encodes microRNA in lytic infections of epithelial cells. BoHV-5-miR-B10 was the most abundant miRNA detected in a high-throughput sequencing study. Here, we evaluated the kinetics of miR-B10 expression after BoHV-5 productive infection by stem-loop real-time quantitative PCR. miR-B10 candidate target sites in the virus were predicted, and BoHV-5 UL39 was confirmed as a target gene by dual-luciferase assay with the design of an miR-B10 tough decoy (TuD). The UL39 gene encoding ribonucleotide reductase (RR) large subunit plays an important role in the early stage of BoHV-5 lytic infection. As BoHV-5-miR-B10 is located in internal and terminal repeat regions, we generated a TuD gene-integrated BoHV-5 strain, which effectively down-regulated miR-B10-3p. Strikingly, the suppression of miR-B10-3p significantly improved BoHV-5 replication. Taking these findings together, our study established an efficient method to deliver and express TuD RNA for viral miRNA suppression, and demonstrated that virus-encoded miRNA suppresses viral-genome biogenesis with a feedback mode, which might serve as a brake for viral replication. Herpesviruses infect humans and a variety of animals. Almost all herpesviruses can encode miRNAs, but the functions of these miRNAs remain to be elucidated. Most herpesvirus-encoded miRNA harbours dual copies, which is difficult to be deleted by current genetic modulation. Here, we developed an efficient method to deliver and express TuD RNA to efficiently suppress viral miRNA with multiple copies. Using this method, we demonstrated for the first time that viral miRNA feedback regulates viral replication by suppressing the expression of RR.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.001375
2020-01-14
2020-01-27
Loading full text...

Full text loading...

References

  1. Roizmann B, Desrosiers RC, Fleckenstein B, Lopez C, Minson AC et al. The family Herpesviridae: an update. The Herpesvirus Study Group of the International Committee on Taxonomy of Viruses. Arch Virol 1992;123:425–449
    [Google Scholar]
  2. Delhon G, Moraes MP, Lu Z, Afonso CL, Flores EF et al. Genome of bovine herpesvirus 5. J Virol 2003;77:10339–10347 [CrossRef]
    [Google Scholar]
  3. Lee BJ, Weiss ML, Mosier D, Chowdhury SI. Spread of bovine herpesvirus type 5 (BHV-5) in the rabbit brain after intranasal inoculation. J Neurovirol 1999;5:474–484 [CrossRef]
    [Google Scholar]
  4. Mesquita LP, Costa RC, Fusuma MM, Bruhn FRP, Mori E et al. Susceptibility of mice to bovine herpesvirus type 5 infection in the central nervous system. Vet Res Commun 2017;41:279–288 [CrossRef]
    [Google Scholar]
  5. Meyer G, Lemaire M, Ros C, Belak K, Gabriel A et al. Comparative pathogenesis of acute and latent infections of calves with bovine herpesvirus types 1 and 5. Arch Virol 2001;146:633–652 [CrossRef]
    [Google Scholar]
  6. Chase CCL, Fulton RW, O’Toole D, Gillette B, Daly RF et al. Bovine herpesvirus 1 modified live virus vaccines for cattle reproduction: balancing protection with undesired effects. Vet Microbiol 2017;206:6977 [CrossRef]
    [Google Scholar]
  7. Jones C, da Silva LF, Sinani D. Regulation of the latency–reactivation cycle by products encoded by the bovine herpesvirus 1 (BHV-1) latency-related gene. J Neurovirol 2011;17:535–545 [CrossRef]
    [Google Scholar]
  8. Simard C, Langlois I, Styger D, Vogt B, Vlcek C et al. Sequence analysis of the UL39, UL38 and UL37 homologues of bovine herpesvirus 1 and expression studies of UL40 and UL39, the subunits of ribonucleotide reductase. Virology 1995;212:734–740 [CrossRef]
    [Google Scholar]
  9. Spear MA, Sun F, Eling DJ, Gilpin E, Kipps TJ et al. Cytotoxicity, apoptosis, and viral replication in tumor cells treated with oncolytic ribonucleotide reductase-defective herpes simplex type 1 virus (hrR3) combined with ionizing radiation. Cancer Gene Ther 2000;7:1051–1059 [CrossRef]
    [Google Scholar]
  10. Cokarić Brdovčak M, Zubković A, Jurak I. Herpes simplex virus 1 deregulation of host microRNAs. ncRNA 2018;4:36 [CrossRef]
    [Google Scholar]
  11. Kincaid RP, Sullivan CS. Virus-Encoded microRNAs: an overview and a look to the future. PLoS Pathog 2012;8:e1003018 [CrossRef]
    [Google Scholar]
  12. Tang Q, Wu Y-Q, Chen D-S, Zhou Q, Chen H-C et al. Bovine herpesvirus 5 encodes a unique pattern of microRNAs compared with bovine herpesvirus 1. J Gen Virol 2014;95:671–678 [CrossRef]
    [Google Scholar]
  13. Glazov EA, Horwood PF, Assavalapsakul W, Kongsuwan K, Mitchell RW et al. Characterization of microRNAs encoded by the bovine herpesvirus 1 genome. J Gen Virol 2010;91:32–41 [CrossRef]
    [Google Scholar]
  14. Kurata JS, Lin R-J. MicroRNA-focused CRISPR-Cas9 library screen reveals fitness-associated miRNAs. RNA 2018;24:966–981 [CrossRef]
    [Google Scholar]
  15. Hollensen AK, Bak RO, Haslund D, Mikkelsen JG. Suppression of microRNAs by dual-targeting and clustered tough decoy inhibitors. RNA Biol 2013;10:406–414 [CrossRef]
    [Google Scholar]
  16. Hutvágner G, Simard MJ, Mello CC, Zamore PD. Sequence-specific inhibition of small RNA function. PLoS Biol 2004;2:e98–475 [CrossRef]
    [Google Scholar]
  17. Meister G, Landthaler M, Dorsett Y, Tuschl T. Sequence-specific inhibition of microRNA- and siRNA-induced RNA silencing. RNA 2004;10:544–550 [CrossRef]
    [Google Scholar]
  18. Elmén J, Lindow M, Schütz S, Lawrence M, Petri A et al. Lna-Mediated microRNA silencing in non-human primates. Nature 2008;452:896–899 [CrossRef]
    [Google Scholar]
  19. Krützfeldt J, Rajewsky N, Braich R, Rajeev KG, Tuschl T et al. Silencing of microRNAs in vivo with ‘antagomirs’. Nature 2005;438:685–689 [CrossRef]
    [Google Scholar]
  20. Ebert MS, Neilson JR, Sharp PA. Microrna sponges: competitive inhibitors of small RNAs in mammalian cells. Nat Methods 2007;4:721–726 [CrossRef]
    [Google Scholar]
  21. Haraguchi T, Ozaki Y, Iba H. Vectors expressing efficient RNA decoys achieve the long-term suppression of specific microRNA activity in mammalian cells. Nucleic Acids Res 2009;37:e43 [CrossRef]
    [Google Scholar]
  22. Xie J, Ameres SL, Friedline R, Hung J-H, Zhang Y et al. Long-term, efficient inhibition of microRNA function in mice using rAAV vectors. Nat Methods 2012;9:403–409 [CrossRef]
    [Google Scholar]
  23. Israelow B, Mullokandov G, Agudo J, Sourisseau M, Bashir A et al. Hepatitis C virus genetics affects miR-122 requirements and response to miR-122 inhibitors. Nat Commun 2014;5: [CrossRef]
    [Google Scholar]
  24. Bak RO, Hollensen AK, Primo MN, Sorensen CD, Mikkelsen JG. Potent microRNA suppression by RNA Pol II-transcribed 'Tough Decoy' inhibitors. RNA 2013;19:280–293 [CrossRef]
    [Google Scholar]
  25. Enright AJ, John B, Gaul U, Tuschl T, Sander C et al. Microrna targets in Drosophila. Genome Biol 2003;5:R1 [CrossRef]
    [Google Scholar]
  26. Pan D, Flores O, Umbach JL, Pesola JM, Bentley P et al. A neuron-specific host microRNA targets herpes simplex virus-1 ICP0 expression and promotes latency. Cell Host Microbe 2014;15:446–456 [CrossRef]
    [Google Scholar]
  27. Boss IW, Plaisance KB, Renne R. Role of virus-encoded microRNAs in herpesvirus biology. Trends Microbiol 2009;17:544–553 [CrossRef]
    [Google Scholar]
  28. Grundhoff A, Sullivan CS. Virus-encoded microRNAs. Virology 2011;411:325–343 [CrossRef]
    [Google Scholar]
  29. Wang X, Zhang M-M, Yan K, Tang Q, Wu Y-Q et al. The full-length microRNA cluster in the intron of large latency transcript is associated with the virulence of pseudorabies virus. Virology 2018;520:5966 [CrossRef]
    [Google Scholar]
  30. Hook LM, Grey F, Grabski R, Tirabassi R, Doyle T et al. Cytomegalovirus miRNAs target secretory pathway genes to facilitate formation of the virion assembly compartment and reduce cytokine secretion. Cell Host Microbe 2014;15:363–373 [CrossRef]
    [Google Scholar]
  31. Liu ZF, Brum MCS, Doster A, Jones C, Chowdhury SI. A bovine herpesvirus type 1 mutant virus specifying a carboxyl-terminal truncation of glycoprotein E is defective in anterograde neuronal transport in rabbits and calves. J Virol 2008;82:7432–7442 [CrossRef]
    [Google Scholar]
  32. Adler H, Messerle M, Wagner M, Koszinowski UH. Cloning and mutagenesis of the murine gammaherpesvirus 68 genome as an infectious bacterial artificial chromosome. J Virol 2000;74:6964–6974 [CrossRef]
    [Google Scholar]
  33. Mostafa HH, Thompson TW, Konen AJ, Haenchen SD, Hilliard JG et al. Herpes simplex virus 1 mutant with point mutations in UL39 is impaired for acute viral replication in mice, establishment of latency, and explant-induced reactivation. J Virol 2018;92: [CrossRef]
    [Google Scholar]
  34. Smith CC. The herpes simplex virus type 2 protein ICP10PK: a master of versatility. Front Biosci 2005;10:2820 [CrossRef]
    [Google Scholar]
  35. Ma W, He H, Wang H. Oncolytic herpes simplex virus and immunotherapy. BMC Immunol 2018;19:40 [CrossRef]
    [Google Scholar]
  36. Kanai R, Zaupa C, Sgubin D, Antoszczyk SJ, Martuza RL et al. Effect of γ34.5 deletions on oncolytic herpes simplex virus activity in brain tumors. J Virol 2012;86:4420–4431 [CrossRef]
    [Google Scholar]
  37. Chowdhury SI, Lee BJ, Mosier D, Sur JH, Osorio FA et al. Neuropathology of bovine herpesvirus type 5 (BHV-5) meningo-encephalitis in a rabbit seizure model. J Comp Pathol 1997;117:295–310 [CrossRef]
    [Google Scholar]
  38. Chen C, Ridzon DA, Broomer AJ, Zhou Z, Lee DH. Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res 2005;33:e179 [CrossRef]
    [Google Scholar]
  39. Wang X, Wu C-X, Song X-R, Chen H-C, Liu Z-F. Comparison of pseudorabies virus China reference strain with emerging variants reveals independent virus evolution within specific geographic regions. Virology 2017;506:9298 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.001375
Loading
/content/journal/jgv/10.1099/jgv.0.001375
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error