1887

Abstract

We examined the influence of inactivation of various genes located in the unique short (U) region of pseudorabies virus on virus replication and assembly in porcine nasal mucosa explant cultures. The following strains were used: the virulent wild-type strain NIA-3, and strains derived from NIA-3 containing a mutation inactivating the genes encoding either the US3-encoded protein kinase (PK), gG, gD, gI, gE, the 28 kDa (‘28K’) protein (single mutant), or the 28K and 11 kDa (‘11K’) proteins (double mutant). In addition a wild-type rescuant was used, which was generated by marker rescue from a PK mutant. All virus strains infected nasal epithelium and had invaded the stroma after approximately 24 h. The morphogenesis in nasal epithelium cells of two PK mutants showed the most striking differences compared to the parent NIA-3 strain and the other mutant strains. The changes could be ascribed to the US3-encoded PK because the rescue mutant showed a similar morphogenesis to wild-type NIA-3. The transmembrane transport of the PK mutants was impaired at the outer nuclear membrane which resulted in an accumulation of virions in the perinuclear space. These results suggest that proteins, phosphorylated by the US3-encoded PK, are involved in debudding of virus particles at the outer nuclear membrane. This defect in the transport of the US3 mutant probably explains their reduced replication . The gG, gD, gI, gE, 28K and 11K mutant strains showed minor or no changes in viral assembly. Thus the reported decreased virulence of the gD, gI and gE mutants was, in contrast to that of the PK mutants, not associated with clear alterations in morphogenesis.

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1995-07-01
2024-03-28
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References

  1. Ben-porat T., Rixon F. J., Blankenship M. L. 1979; Analysis of the structure of the genome of pseudorabies virus. Virology 95:285–294
    [Google Scholar]
  2. Campadelli-fiume G., Farabegoli F., Di gaeta S., Roizman B. 1991; Origin of unenveloped capsids in the cytoplasm of cells infected with herpes simplex virus 1. Journal of Virology 65:1589–1595
    [Google Scholar]
  3. Daikoku T., Kurachi R., Tsurumi T., Nishiyama Y. 1994; Identification of a target protein of US3 protein kinase of herpes simplex virus type 2. Journal of General Virology 75:2065–2068
    [Google Scholar]
  4. De wind N., Zijderveld A., Glazenburg K., Gielkens A., Berns A. 1990; Linker insertion mutagenesis of herpesviruses: inactivation of single genes within the Us region of pseudorabies virus. Journal of Virology 64:4691–4696
    [Google Scholar]
  5. De wind N., Domen J., Berns A. 1992; Herpesviruses encode an unusual protein – serine/threonine kinase which is nonessential for growth in cultured cells. Journal of Virology 66:5200–5209
    [Google Scholar]
  6. Eggers M., Bogner E., Agricola B., Kern H. F., Radsak K. 1991; Inhibition of human cytomegalovirus maturation by bre–feldin A. Journal of General Virology 73:2679–2692
    [Google Scholar]
  7. Frame M. C., Purves F. C., Mcgeoch D. J., Marsden H. S., Leader D. P. 1987; Identification of the herpes simplex virus protein kinase as the product of viral gene US3. Journal of General Virology 68:2699–2704
    [Google Scholar]
  8. Gibson W., Roizman B. 1972; Proteins specified by herpes simplex virus. VIII. Characterization and composition of multiple capsid forms of subtypes 1 and 2. Journal of Virology 10:1044–1052
    [Google Scholar]
  9. Jacobs L., Rziha H. J., Kimman T. G., Gielkens A. L. J., Van oirschot J. T. 1993; Deleting valine-125 and cysteine-126 in glycoprotein gl of pseudorabies virus strain NIA-3 decreases plaque size and reduces virulence in mice. Archives of Virology 131:251–264
    [Google Scholar]
  10. Katsumoto T., Hirano A., Kurimura T., Takagi A. 1981; In situ electron microscopical observation of cells infected with herpes simplex virus. Journal of General Virology 52:267–278
    [Google Scholar]
  11. Kimman T. G., De wind N., Oei-lie N., Pol J. M. A., Berns A. J. M., Gielkens A. L. J. 1992; Contribution of single genes within the unique short region of Aujeszky’s disease virus (suid herpesvirus type 1) to virulence, pathogenesis and immunogenicity. Journal of General Virology 13:243–251
    [Google Scholar]
  12. Kimman T. G., De wind N., De bruin T., De visser Y., Voermans J. 1994; Inactivation of glycoprotein gE and thymidine kinase or the US3-encoded protein kinase synergistically decreases in vivo replication of pseudorabies virus and the induction of protective immunity. Virology 205:511–518
    [Google Scholar]
  13. Kritas S. K., Pensaert M. B., Mettenleiter T. C. 1994a; Invasion and spread of single glycoprotein deleted mutants of Aujeszky’s disease virus (ADV) in the trigeminal nervous pathway of pigs after intranasal inoculation. Veterinary Microbiology 40:323–334
    [Google Scholar]
  14. Kritas S. K., Pensaert M. B., Mettenleiter T. C. 1994b; Role of envelope glycoproteins gl, gp63 and gill in the invasion and spread of Aujeszky’s disease virus in the olfactory nervous pathway of the pig. Journal of General Virology 75:2319–2327
    [Google Scholar]
  15. Kurachi R., Daikoku T., Tsurumr T., Maeno K., Nishiyama K., Kurata T. 1993; The pathogenicity of a US3 protein kinase-deficient mutant of herpes simplex type 2 in mice. Archives of Virology 133:259–273
    [Google Scholar]
  16. Mettenleiter T. C., Zsak L., Kaplan A. S., Ben-porat T., Lomnici B. 1987; Role of a structural glycoprotein of pseudorabies virus in virus virulence. Journal of Virology 61:4030–4032
    [Google Scholar]
  17. Mulder W. A. M., Jacobs L., Priem J., Kok G. L., Wagenaar F., Kjmman T. G., Pol J. M. A. 1994; Glycoprotein gE-negative pseudorabies virus has a reduced capability to infect second- and third-order neurons of the olfactory and trigeminal routes in the porcine central nervous system. Journal of General Virology 75:3095–3106
    [Google Scholar]
  18. Nishiyama Y., Yamada Y., Kurachi R., Daikoku T. 1992; Construction of a US3 lac Z insertion mutant of herpes simplex virus type 2 and characterization of its phenotype in vitro and in vivo. Virology 190:256–268
    [Google Scholar]
  19. Peeters B., De wind N., Hooisma M., Wagenaar F., Gielkens A., Moormann R. 1992; Pseudorabies virus envelope glycoproteins gp50 and gll are essential for virus penetration, but only gII is involved in membrane fusion. Journal of Virology 66:894–905
    [Google Scholar]
  20. Peeters B., Pol J., Gielkens A., Moormann R. 1993; Envelope glycoprotein gp50 of pseudorabies virus is essential for virus entry but is not required for viral spread in mice. Journal of Virology 67:170–177
    [Google Scholar]
  21. Peeters B., Bouma A., De bruin T., Moormann R., Gielkens A., Kimman T. 1994; Non-transmissible pseudorabies virus gp50 mutants: a new generation of safe live vaccines. Vaccine 12:375–380
    [Google Scholar]
  22. Petrovskis E. A., Post L. E. 1987; A small open reading frame in pseudorabies virus and implications for evolutionary relationships between herpesviruses. Virology 159:193–195
    [Google Scholar]
  23. Petrovskis E. A., Timmens J. G., Post L. E. 1986a; Use of lambda gt11 to isolate genes for two pseudorabies virus glycoproteins with homology to herpes simplex virus and varicella-zoster virus glycoproteins. Journal of Virology 60:185–193
    [Google Scholar]
  24. Petrovskis E. A., Timmens J. G., Armentrout M. A., Marchioli C. C., Yancey R. J. Jr, Post L. E. 1986b; DNA sequence of the gene for pseudorabiesvirus gp50, a glycoprotein without N-linked glycosylation. Journal of Virology 59:216–223
    [Google Scholar]
  25. Pol J. M. A., Quint W. G. V., Kok G. L., Broekhuijsen-davies J. M. 1991a; Pseudorabies virus infections in explants of porcine nasal mucosa. Research in Veterinary Science 50:45–53
    [Google Scholar]
  26. Pol J. M. A., Wagenaar F., Gielkens A. 1991b; Morphogenesis of three pseudorabies virus strains in porcine nasal mucosa. Intervirology 32:327–337
    [Google Scholar]
  27. Purves F. C., Spector D., Roizman B. 1991; The herpes simplex virus 1 protein kinase encoded by the US3 gene mediates posttranslational modification of the phosphoprotein encoded by the UL34 gene. Journal of Virology 65:5757–5764
    [Google Scholar]
  28. Rea T. J., Timmens J. G., Long G. W., Post L. E. 1985; Mapping and sequence of the gene for the pseudorabies virus glycoprotein which accumulates in the medium of infected cells. Journal of Virology 54:21–29
    [Google Scholar]
  29. Roizman B., Sears A. 1990; Flerpes simplex viruses and their replication. In Virology 2nd edn Edited by Fields B. N., Knipe M. D. pp 1795–1841 New York: Raven Press;
    [Google Scholar]
  30. Thomsen D. R., Marchioli C. C., Yancey R. J. Jr, Post L. E. 1987; Replication and virulence of pseudorabies virus mutants lacking glycoprotein gX. Journal of Virology 61:229–232
    [Google Scholar]
  31. Van zijl M., Quint W., Briaire J., De rover T., Gielkens A., Berns A. 1988; Regeneration of herpesviruses from molecularly cloned subgenomic fragments. Journal of Virology 62:2191–2195
    [Google Scholar]
  32. Van zijl M., Van der gulden H., De wind N., Gielkens A., Berns A. 1990; Identification of two genes in the unique short region of pseudorabies virus; comparison with herpes simplex virus and varicella-zoster virus. Journal of General Virology 71:1747–1755
    [Google Scholar]
  33. Wagenaar F., Kok G. L., Broekhuijsen-davies J. M., Pol J. M. A. 1993; Rapid cold fixation of tissue samples by microwave irradiation for use in electron microscopy. Histochemical Journal 25:719–725
    [Google Scholar]
  34. Whealy M. E., Card J. P., Meade R. P., Robbins A. K., Enquist L. W. 1991; Effect of brefeldin A on alphaherpesvirus membrane protein glycosylation and virus egress. Journal of Virology 65:1066–1081
    [Google Scholar]
  35. Whealy M. E., Card J. P., Robbins A. K., Dubin J. R., Rziha H.-J., Enquist L. W. 1993; Specific pseudorabies virus infection of the rat visual system requires both gl and gp63 glycoproteins. Journal of Virology 67:3786–3797
    [Google Scholar]
  36. Zhang G., Stevens R., Leader D. P. 1990; The protein kinase encoded in the short unique region of pseudorabies virus: description of the gene and identification of its product in virions and in infected cells. Journal of General Virology 711757–1765
    [Google Scholar]
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