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. 1997 Jan;71(1):17–24. doi: 10.1128/jvi.71.1.17-24.1997

Adaptability in herpesviruses: glycoprotein D-independent infectivity of pseudorabies virus.

J Schmidt 1, B G Klupp 1, A Karger 1, T C Mettenleiter 1
PMCID: PMC191019  PMID: 8985318

Abstract

Initial contact between herpesviruses and host cells is mediated by virion envelope glycoproteins which bind to cellular receptors. In several alphaherpesviruses, the nonessential glycoprotein gC has been found to interact with cell surface proteoglycans, whereas the essential glycoprotein gD is involved in stable secondary attachment. In addition, gD is necessary for penetration, which involves fusion between virion envelope and cellular cytoplasmic membrane. As opposed to other alphaherpesvirus gD homologs, pseudorabies virus (PrV) gD is not required for direct viral cell-to-cell spread. Therefore, gD- PrV can be passaged in noncomplementing cells by cocultivating infected and noninfected cells. Whereas infectivity was found to be strictly cell associated in early passages, repeated passaging resulted in the appearance of infectivity in the supernatant, finally reaching titers as high as 10(7) PFU/ml (PrV gD- Pass). Filtration experiments indicated that this infectivity was not due to the presence of infected cells, and the absence of gD was verified by Southern and Western blotting and by virus neutralization. Infection of bovine kidney cells constitutively expressing PrV gD interfered with the infectivity of wild-type PrV but did not inhibit that of PrV gD- Pass. Similar results were obtained after passaging of a second PrV mutant, PrV-376, which in addition to gD also lacks gG, gI, and gE. Penetration assays demonstrated that PrV gD- Pass entered cells much more slowly than wild-type PrV. In summary, our data demonstrate the existence of a gD-independent mode of initiation of infection in PrV and indicate that the essential function(s) that gD performs in wild-type PrV infection can be compensated for after passaging. Therefore, regarding the requirement for gD, PrV seems to be intermediate between herpes simplex virus type 1, in which gD is necessary for penetration and cell-to-cell spread, and varicella-zoster virus (VZV), which lacks a gD gene. Our data show that the relevance of an essential protein can change under selective pressure and thus demonstrate a way in which VZV could have evolved from a PrV-like ancestor.

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Selected References

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  1. Brunetti C. R., Burke R. L., Kornfeld S., Gregory W., Masiarz F. R., Dingwell K. S., Johnson D. C. Herpes simplex virus glycoprotein D acquires mannose 6-phosphate residues and binds to mannose 6-phosphate receptors. J Biol Chem. 1994 Jun 24;269(25):17067–17074. [PubMed] [Google Scholar]
  2. Buckmaster A. E., Scott S. D., Sanderson M. J., Boursnell M. E., Ross N. L., Binns M. M. Gene sequence and mapping data from Marek's disease virus and herpesvirus of turkeys: implications for herpesvirus classification. J Gen Virol. 1988 Aug;69(Pt 8):2033–2042. doi: 10.1099/0022-1317-69-8-2033. [DOI] [PubMed] [Google Scholar]
  3. Campadelli-Fiume G., Arsenakis M., Farabegoli F., Roizman B. Entry of herpes simplex virus 1 in BJ cells that constitutively express viral glycoprotein D is by endocytosis and results in degradation of the virus. J Virol. 1988 Jan;62(1):159–167. doi: 10.1128/jvi.62.1.159-167.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Campadelli-Fiume G., Qi S., Avitabile E., Foà-Tomasi L., Brandimarti R., Roizman B. Glycoprotein D of herpes simplex virus encodes a domain which precludes penetration of cells expressing the glycoprotein by superinfecting herpes simplex virus. J Virol. 1990 Dec;64(12):6070–6079. doi: 10.1128/jvi.64.12.6070-6079.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chase C. C., Carter-Allen K., Lohff C., Letchworth G. J., 3rd Bovine cells expressing bovine herpesvirus 1 (BHV-1) glycoprotein IV resist infection by BHV-1, herpes simplex virus, and pseudorabies virus. J Virol. 1990 Oct;64(10):4866–4872. doi: 10.1128/jvi.64.10.4866-4872.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Davison A. J., Scott J. E. The complete DNA sequence of varicella-zoster virus. J Gen Virol. 1986 Sep;67(Pt 9):1759–1816. doi: 10.1099/0022-1317-67-9-1759. [DOI] [PubMed] [Google Scholar]
  7. Dean H. J., Terhune S. S., Shieh M. T., Susmarski N., Spear P. G. Single amino acid substitutions in gD of herpes simplex virus 1 confer resistance to gD-mediated interference and cause cell-type-dependent alterations in infectivity. Virology. 1994 Feb 15;199(1):67–80. doi: 10.1006/viro.1994.1098. [DOI] [PubMed] [Google Scholar]
  8. Dean H. J., Warner M. S., Terhune S. S., Johnson R. M., Spear P. G. Viral determinants of the variable sensitivity of herpes simplex virus strains to gD-mediated interference. J Virol. 1995 Aug;69(8):5171–5176. doi: 10.1128/jvi.69.8.5171-5176.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fuchs W., Klupp B. G., Granzow H., Rziha H. J., Mettenleiter T. C. Identification and characterization of the pseudorabies virus UL3.5 protein, which is involved in virus egress. J Virol. 1996 Jun;70(6):3517–3527. doi: 10.1128/jvi.70.6.3517-3527.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  11. Hanssens F. P., Nauwynck H. J., Pensaert M. B. Involvement of membrane-bound viral glycoproteins in adhesion of pseudorabies virus-infected cells. J Virol. 1993 Aug;67(8):4492–4496. doi: 10.1128/jvi.67.8.4492-4496.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Heffner S., Kovács F., Klupp B. G., Mettenleiter T. C. Glycoprotein gp50-negative pseudorabies virus: a novel approach toward a nonspreading live herpesvirus vaccine. J Virol. 1993 Mar;67(3):1529–1537. doi: 10.1128/jvi.67.3.1529-1537.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Isfort R. J., Qian Z., Jones D., Silva R. F., Witter R., Kung H. J. Integration of multiple chicken retroviruses into multiple chicken herpesviruses: herpesviral gD as a common target of integration. Virology. 1994 Aug 15;203(1):125–133. doi: 10.1006/viro.1994.1462. [DOI] [PubMed] [Google Scholar]
  14. Johnson D. C., Burke R. L., Gregory T. Soluble forms of herpes simplex virus glycoprotein D bind to a limited number of cell surface receptors and inhibit virus entry into cells. J Virol. 1990 Jun;64(6):2569–2576. doi: 10.1128/jvi.64.6.2569-2576.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Johnson R. M., Spear P. G. Herpes simplex virus glycoprotein D mediates interference with herpes simplex virus infection. J Virol. 1989 Feb;63(2):819–827. doi: 10.1128/jvi.63.2.819-827.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. KAPLAN A. S., VATTER A. E. A comparison of herpes simplex and pseudorabies viruses. Virology. 1959 Apr;7(4):394–407. doi: 10.1016/0042-6822(59)90068-6. [DOI] [PubMed] [Google Scholar]
  17. Karger A., Mettenleiter T. C. Glycoproteins gIII and gp50 play dominant roles in the biphasic attachment of pseudorabies virus. Virology. 1993 Jun;194(2):654–664. doi: 10.1006/viro.1993.1305. [DOI] [PubMed] [Google Scholar]
  18. Karger A., Saalmüller A., Tufaro F., Banfield B. W., Mettenleiter T. C. Cell surface proteoglycans are not essential for infection by pseudorabies virus. J Virol. 1995 Jun;69(6):3482–3489. doi: 10.1128/jvi.69.6.3482-3489.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kimman T. G., de Wind N., Oei-Lie N., Pol J. M., Berns A. J., Gielkens A. L. Contribution of single genes within the unique short region of Aujeszky's disease virus (suid herpesvirus type 1) to virulence, pathogenesis and immunogenicity. J Gen Virol. 1992 Feb;73(Pt 2):243–251. doi: 10.1099/0022-1317-73-2-243. [DOI] [PubMed] [Google Scholar]
  20. Klupp B. G., Baumeister J., Karger A., Visser N., Mettenleiter T. C. Identification and characterization of a novel structural glycoprotein in pseudorabies virus, gL. J Virol. 1994 Jun;68(6):3868–3878. doi: 10.1128/jvi.68.6.3868-3878.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Klupp B. G., Visser N., Mettenleiter T. C. Identification and characterization of pseudorabies virus glycoprotein H. J Virol. 1992 May;66(5):3048–3055. doi: 10.1128/jvi.66.5.3048-3055.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kopp A., Mettenleiter T. C. Stable rescue of a glycoprotein gII deletion mutant of pseudorabies virus by glycoprotein gI of bovine herpesvirus 1. J Virol. 1992 May;66(5):2754–2762. doi: 10.1128/jvi.66.5.2754-2762.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  24. Liang X., Pyne C., Li Y., Babiuk L. A., Kowalski J. Delineation of the essential function of bovine herpesvirus 1 gD: an indication for the modulatory role of gD in virus entry. Virology. 1995 Mar 10;207(2):429–441. doi: 10.1006/viro.1995.1102. [DOI] [PubMed] [Google Scholar]
  25. Ligas M. W., Johnson D. C. A herpes simplex virus mutant in which glycoprotein D sequences are replaced by beta-galactosidase sequences binds to but is unable to penetrate into cells. J Virol. 1988 May;62(5):1486–1494. doi: 10.1128/jvi.62.5.1486-1494.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lukàcs N., Thiel H. J., Mettenleiter T. C., Rziha H. J. Demonstration of three major species of pseudorabies virus glycoproteins and identification of a disulfide-linked glycoprotein complex. J Virol. 1985 Jan;53(1):166–173. doi: 10.1128/jvi.53.1.166-173.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. McClain D. S., Fuller A. O. Cell-specific kinetics and efficiency of herpes simplex virus type 1 entry are determined by two distinct phases of attachment. Virology. 1994 Feb;198(2):690–702. doi: 10.1006/viro.1994.1081. [DOI] [PubMed] [Google Scholar]
  28. McGeoch D. J. Evolutionary relationships of virion glycoprotein genes in the S regions of alphaherpesvirus genomes. J Gen Virol. 1990 Oct;71(Pt 10):2361–2367. doi: 10.1099/0022-1317-71-10-2361. [DOI] [PubMed] [Google Scholar]
  29. Mettenleiter T. C. Glycoprotein gIII deletion mutants of pseudorabies virus are impaired in virus entry. Virology. 1989 Aug;171(2):623–625. doi: 10.1016/0042-6822(89)90635-1. [DOI] [PubMed] [Google Scholar]
  30. Mettenleiter T. C. Initiation and spread of alpha-herpesvirus infections. Trends Microbiol. 1994 Jan;2(1):2–4. doi: 10.1016/0966-842x(94)90335-2. [DOI] [PubMed] [Google Scholar]
  31. Mettenleiter T. C., Klupp B. G., Weiland F., Visser N. Characterization of a quadruple glycoprotein-deleted pseudorabies virus mutant for use as a biologically safe live virus vaccine. J Gen Virol. 1994 Jul;75(Pt 7):1723–1733. doi: 10.1099/0022-1317-75-7-1723. [DOI] [PubMed] [Google Scholar]
  32. Mettenleiter T. C. Pseudorabies (Aujeszky's disease) virus: state of the art. August 1993. Acta Vet Hung. 1994;42(2-3):153–177. [PubMed] [Google Scholar]
  33. Mettenleiter T. C., Rauh I. A glycoprotein gX-beta-galactosidase fusion gene as insertional marker for rapid identification of pseudorabies virus mutants. J Virol Methods. 1990 Oct;30(1):55–65. doi: 10.1016/0166-0934(90)90043-f. [DOI] [PubMed] [Google Scholar]
  34. Parcells M. S., Anderson A. S., Morgan R. W. Characterization of a Marek's disease virus mutant containing a lacZ insertion in the US6 (gD) homologue gene. Virus Genes. 1994 Sep;9(1):5–13. doi: 10.1007/BF01703430. [DOI] [PubMed] [Google Scholar]
  35. Peeters B., Bouma A., de Bruin T., Moormann R., Gielkens A., Kimman T. Non-transmissible pseudorabies virus gp50 mutants: a new generation of safe live vaccines. Vaccine. 1994 Mar;12(4):375–380. doi: 10.1016/0264-410x(94)90104-x. [DOI] [PubMed] [Google Scholar]
  36. Peeters B., de Wind N., Hooisma M., Wagenaar F., Gielkens A., Moormann R. Pseudorabies virus envelope glycoproteins gp50 and gII are essential for virus penetration, but only gII is involved in membrane fusion. J Virol. 1992 Feb;66(2):894–905. doi: 10.1128/jvi.66.2.894-905.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Petrovskis E. A., Meyer A. L., Post L. E. Reduced yield of infectious pseudorabies virus and herpes simplex virus from cell lines producing viral glycoprotein gp50. J Virol. 1988 Jun;62(6):2196–2199. doi: 10.1128/jvi.62.6.2196-2199.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rauh I., Mettenleiter T. C. Pseudorabies virus glycoproteins gII and gp50 are essential for virus penetration. J Virol. 1991 Oct;65(10):5348–5356. doi: 10.1128/jvi.65.10.5348-5356.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Rauh I., Weiland F., Fehler F., Keil G. M., Mettenleiter T. C. Pseudorabies virus mutants lacking the essential glycoprotein gII can be complemented by glycoprotein gI of bovine herpesvirus 1. J Virol. 1991 Feb;65(2):621–631. doi: 10.1128/jvi.65.2.621-631.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Roizman B. The structure and isomerization of herpes simplex virus genomes. Cell. 1979 Mar;16(3):481–494. doi: 10.1016/0092-8674(79)90023-0. [DOI] [PubMed] [Google Scholar]
  41. Schröder C., Linde G., Fehler F., Keil G. M. From essential to beneficial: glycoprotein D loses importance for replication of bovine herpesvirus 1 in cell culture. J Virol. 1997 Jan;71(1):25–33. doi: 10.1128/jvi.71.1.25-33.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Zsak L., Sugg N., Ben-Porat T. The different interactions of a gIII mutant of pseudorabies virus with several different cell types. J Gen Virol. 1992 Apr;73(Pt 4):821–827. doi: 10.1099/0022-1317-73-4-821. [DOI] [PubMed] [Google Scholar]

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