1887

Abstract

Bovine herpesvirus 1 (BHV-1) infection may lead to conjunctivitis, upper respiratory tract problems, pneumonia, genital disorders and abortion. BHV-1 is able to spread quickly in a plaque-wise manner and invade by breaching the basement membrane (BM) barrier in the respiratory mucosa. BHV-1 Us3, a serine/threonine kinase, induces a dramatic cytoskeletal reorganization and BHV-1 Us9, a tail-anchored membrane protein, is required for axonal transport of viruses in neurons. In this study, we investigated the role of Us3 and Us9 during BHV-1 infection in the respiratory mucosa. First, we constructed and characterized BHV-1 Us3 null, Us9 null and revertant viruses. Then, we analysed the viral replication and plaque size (latitude) in Madin–Darby bovine kidney (MDBK) cells and the respiratory mucosa as well as viral penetration depth underneath the BM of the respiratory mucosa when inoculated with these recombinant viruses. Knockout of Us3 resulted in a 1 log reduction in viral titre and plaque size (latitude) in MDBK cells and the trachea mucosa. There were no defects in the cell-to-cell spread observed for BHV-1 Us9 null virus. Both BHV-1 Us3 null and Us9 null viruses showed a significant reduction of plaque penetration underneath the BM; however, penetration was not completely inhibited. In conclusion, the current findings demonstrated that Us3 and Us9 play an important role in the invasion of BHV-1 through the BM of the respiratory mucosa, which shows the way forward for research-based attenuation of viruses in order to make safer and better-performing vaccines.

Keyword(s): BHV-1 , respiratory mucosa , Us3 and Us9
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2017-05-01
2024-12-04
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References

  1. Jones C, Chowdhury S. Bovine herpesvirus type 1 (BHV-1) is an important cofactor in the bovine respiratory disease complex. Vet Clin North Am Food Anim Pract 2010; 26:303–321 [View Article][PubMed]
    [Google Scholar]
  2. Tikoo SK, Campos M, Babiuk LA. Bovine herpesvirus 1 (BHV-1): biology, pathogenesis, and control. Adv Virus Res 1995; 45:191–223[PubMed] [CrossRef]
    [Google Scholar]
  3. Muylkens B, Thiry J, Kirten P, Schynts F, Thiry E. Bovine herpesvirus 1 infection and infectious bovine rhinotracheitis. Vet Res 2007; 38:181–209 [View Article][PubMed]
    [Google Scholar]
  4. Steukers L, Vandekerckhove AP, Van Den Broeck W, Glorieux S, Nauwynck HJ. Comparative analysis of replication characteristics of BoHV-1 subtypes in bovine respiratory and genital mucosa explants: a phylogenetic enlightenment. Vet Res 2011; 42:33 [View Article]
    [Google Scholar]
  5. Brzozowska A, Rychłowski M, Lipińska AD, Bieńkowska-Szewczyk K. Point mutations in BHV-1 Us3 gene abolish its ability to induce cytoskeletal changes in various cell types. Vet Microbiol 2010; 143:8–13 [View Article][PubMed]
    [Google Scholar]
  6. Steukers L. Nulla tenaci invia est via: unraveling herpesvirus mucosal invasion in an ex vivo organ culture PhD thesis, Ghent University 2013
    [Google Scholar]
  7. Klupp BG, Granzow H, Mettenleiter TC. Effect of the pseudorabies virus US3 protein on nuclear membrane localization of the UL34 protein and virus egress from the nucleus. J Gen Virol 2001; 82:2363–2371 [View Article][PubMed]
    [Google Scholar]
  8. Benetti L, Roizman B. Herpes simplex virus protein kinase US3 activates and functionally overlaps protein kinase A to block apoptosis. Proc Natl Acad Sci USA 2004; 101:9411–9416 [View Article][PubMed]
    [Google Scholar]
  9. Murata T, Goshima F, Yamauchi Y, Koshizuka T, Takakuwa H et al. Herpes simplex virus type 2 US3 blocks apoptosis induced by sorbitol treatment. Microbes Infect 2002; 4:707–712 [View Article][PubMed]
    [Google Scholar]
  10. Takashima Y, Tamura H, Xuan X, Otsuka H. Identification of the US3 gene product of BHV-1 as a protein kinase and characterization of BHV-1 mutants of the US3 gene. Virus Res 1999; 59:23–34 [View Article][PubMed]
    [Google Scholar]
  11. Brideau AD, Card JP, Enquist LW. Role of pseudorabies virus Us9, a type II membrane protein, in infection of tissue culture cells and the rat nervous system. J Virol 2000; 74:834–845 [View Article][PubMed]
    [Google Scholar]
  12. Lyman MG, Kemp CD, Taylor MP, Enquist LW. Comparison of the pseudorabies virus Us9 protein with homologs from other veterinary and human alphaherpesviruses. J Virol 2009; 83:6978–6986 [View Article][PubMed]
    [Google Scholar]
  13. Chowdhury SI, Mahmood S, Simon J, Al-Mubarak A, Zhou Y. The Us9 gene of bovine herpesvirus 1 (BHV-1) effectively complements a Us9-null strain of BHV-5 for anterograde transport, neurovirulence, and neuroinvasiveness in a rabbit model. J Virol 2006; 80:4396–4405 [View Article][PubMed]
    [Google Scholar]
  14. Butchi N, Jones C, Perez S, Doster A, Chowdhury S. Role of envelope protein Us9 in the anterograde transport of bovine herpesvirus-1 (BHV-1) following reactivation in the trigeminal ganglia. J Neurovirol 2007; 13:384–388 [CrossRef]
    [Google Scholar]
  15. Arsenakis M, Campadelli-Fiume G, Lombardo MT, Roizman B. The glycoprotein C gene of herpes simplex virus 1 resident in clonal L cell lines manifests two regulatory domains conferring a dominant B and a subordinate γ2 regulation. Virology 1988; 162:300–310 [View Article][PubMed]
    [Google Scholar]
  16. Liman A, Engels M, Meyer G, Ackermann M. Glycoprotein C of bovine herpesvirus 5 (BHV-5) confers a distinct heparin-binding phenotype to BHV-1. Arch Virol 2000; 145:2047–2059 [View Article][PubMed]
    [Google Scholar]
  17. Kaashoek MJ, Rijsewijk FA, Ruuls RC, Keil GM, Thiry E et al. Virulence, immunogenicity and reactivation of bovine herpesvirus 1 mutants with a deletion in the gC, gG, gI, gE, or in both the gI and gE gene. Vaccine 1998; 16:802–809 [View Article][PubMed]
    [Google Scholar]
  18. Deruelle MJ, Favoreel HW. Keep it in the subfamily: the conserved alphaherpesvirus US3 protein kinase. J Gen Virol 2011; 92:18–30 [View Article][PubMed]
    [Google Scholar]
  19. Lyman MG, Feierbach B, Curanovic D, Bisher M, Enquist LW. Pseudorabies virus Us9 directs axonal sorting of viral capsids. J Virol 2007; 81:11363–11371 [View Article][PubMed]
    [Google Scholar]
  20. Haugo AC, Szpara ML, Parsons L, Enquist LW, Roller RJ. Herpes simplex virus 1 pUL34 plays a critical role in cell-to-cell spread of virus in addition to its role in virus replication. J Virol 2011; 85:7203–7215 [View Article][PubMed]
    [Google Scholar]
  21. Coats CJ. Development of primary neuronal culture of embryonic rabbit dorsal root ganglia for microfluidic chamber analysis of axon mediated neuronal spread of bovine herpesvirus type 1 PhD thesis, Kansas State University 2010
    [Google Scholar]
  22. Lamote JA, Glorieux S, Nauwynck HJ, Favoreel HW. The US3 protein of pseudorabies virus drives viral passage across the basement membrane in porcine respiratory mucosa explants. J Virol 2016; 90:10945–10950 [View Article][PubMed]
    [Google Scholar]
  23. Chuluunbaatar U, Mohr I. A herpesvirus kinase that masquerades as Akt: you don't have to look like Akt, to act like it. Cell Cycle 2011; 10:2064–2068 [View Article][PubMed]
    [Google Scholar]
  24. Sun X, Gao X, Zhou L, Sun L, Lu C. PDGF-BB-induced MT1-MMP expression regulates proliferation and invasion of mesenchymal stem cells in 3-dimensional collagen via MEK/ERK1/2 and PI3K/AKT signaling. Cell Signal 2013; 25:1279–1287 [View Article][PubMed]
    [Google Scholar]
  25. Labiuk SL, Babiuk LA, van Drunen Littel-van den Hurk S. Major tegument protein VP8 of bovine herpesvirus 1 is phosphorylated by viral US3 and cellular CK2 protein kinases. J Gen Virol 2009; 90:2829–2839 [View Article][PubMed]
    [Google Scholar]
  26. Labiuk SL, Lobanov V, Lawman Z, Snider M, Babiuk LA et al. Bovine herpesvirus-1 US3 protein kinase: critical residues and involvement in the phosphorylation of VP22. J Gen Virol 2010; 91:1117–1126 [View Article][PubMed]
    [Google Scholar]
  27. Jiang YM, Yamada H, Goshima F, Daikoku T, Oshima S et al. Characterization of the herpes simplex virus type 2 (HSV-2) US2 gene product and a US2-deficient HSV-2 mutant. J Gen Virol 1998; 79:2777–2784 [View Article][PubMed]
    [Google Scholar]
  28. Kato A, Yamamoto M, Ohno T, Kodaira H, Nishiyama Y et al. Identification of proteins phosphorylated directly by the Us3 protein kinase encoded by herpes simplex virus 1. J Virol 2005; 79:9325–9331 [View Article][PubMed]
    [Google Scholar]
  29. Matsuzaki A, Yamauchi Y, Kato A, Goshima F, Kawaguchi Y et al. US3 protein kinase of herpes simplex virus type 2 is required for the stability of the UL46-encoded tegument protein and its association with virus particles. J Gen Virol 2005; 86:1979–1985 [View Article][PubMed]
    [Google Scholar]
  30. Leopardi R, van Sant C, Roizman B. The herpes simplex virus 1 protein kinase US3 is required for protection from apoptosis induced by the virus. Proc Natl Acad Sci USA 1997; 94:7891–7896 [View Article][PubMed]
    [Google Scholar]
  31. Geenen K, Favoreel HW, Olsen L, Enquist LW, Nauwynck HJ. The pseudorabies virus US3 protein kinase possesses anti-apoptotic activity that protects cells from apoptosis during infection and after treatment with sorbitol or staurosporine. Virology 2005; 331:144–150 [View Article][PubMed]
    [Google Scholar]
  32. Young LS, Dawson CW, Eliopoulos AG. Viruses and apoptosis. Br Med Bull 1997; 53:509–521 [View Article][PubMed]
    [Google Scholar]
  33. Al-Mubarak A, Simon J, Coats C, Okemba JD, Burton MD et al. Glycoprotein E (gE) specified by bovine herpesvirus type 5 (BHV-5) enables trans-neuronal virus spread and neurovirulence without being a structural component of enveloped virions. Virology 2007; 365:398–409 [View Article][PubMed]
    [Google Scholar]
  34. Kramer T, Greco TM, Taylor MP, Ambrosini AE, Cristea IM et al. Kinesin-3 mediates axonal sorting and directional transport of alphaherpesvirus particles in neurons. Cell Host Microbe 2012; 12:806–814 [View Article][PubMed]
    [Google Scholar]
  35. Kratchmarov R, Kramer T, Greco TM, Taylor MP, Ch'ng TH et al. Glycoproteins gE and gI are required for efficient KIF1A-dependent anterograde axonal transport of alphaherpesvirus particles in neurons. J Virol 2013; 87:9431–9440 [View Article][PubMed]
    [Google Scholar]
  36. Ch'ng TH, Enquist LW. Efficient axonal localization of alphaherpesvirus structural proteins in cultured sympathetic neurons requires viral glycoprotein E. J Virol 2005; 79:8835–8846 [View Article][PubMed]
    [Google Scholar]
  37. Awasthi S, Friedman HM. Molecular association of herpes simplex virus type 1 glycoprotein E with membrane protein Us9. Arch Virol 2016; 161:3203–3213 [View Article][PubMed]
    [Google Scholar]
  38. Liu ZF, Brum MC, 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 [View Article][PubMed]
    [Google Scholar]
  39. Tischer BK, Smith GA, Osterrieder N. En passant mutagenesis: a two step markerless Red recombination system. Methods Mol Biol 2010; 634:421–430 [View Article][PubMed]
    [Google Scholar]
  40. Chowdhury SI. Molecular basis of antigenic variation between the glycoproteins C of respiratory bovine herpesvirus 1 (BHV-1) and neurovirulent BHV-5. Virology 1995; 213:558–568 [View Article][PubMed]
    [Google Scholar]
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