Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1993 Mar;67(3):1236–1245. doi: 10.1128/jvi.67.3.1236-1245.1993

Pseudorabies virus protein homologous to herpes simplex virus type 1 ICP18.5 is necessary for capsid maturation.

T C Mettenleiter 1, A Saalmüller 1, F Weiland 1
PMCID: PMC237489  PMID: 8382292

Abstract

In pseudorabies virus (PrV), an open reading frame that partially overlaps the gene for the essential glycoprotein gII has been shown to encode a protein homologous to the ICP18.5 polypeptide of herpes simplex virus type 1 (N. Pederson and L. Enquist, Nucleic Acids Res. 17:3597, 1989). To study the function of this protein during the viral replicative cycle, a PrV mutant which carries a beta-galactosidase expression cassette interrupting the ICP18.5(PrV) gene was constructed. This mutant could be propagated only on cell lines that were able to provide ICP18.5(PrV) in trans after transformation with a corresponding genomic PrV DNA fragment. Detailed analysis showed that inactivation of the ICP18.5(PrV) gene did not impair infection of noncomplementing cells, nor did it impair early or late gene expression, as shown by immunoprecipitation of glycoproteins gII, gIII, and gp50. Surface localization of glycoproteins as demonstrated by fluorescence-activated cell sorting analyses was also not affected. Southern blot hybridizations, however, showed that cleavage of replicative concatemeric viral DNA did not occur in noncomplementing cells infected by the ICP18.5 mutant PrV. In addition, electron microscopic analysis revealed an accumulation of empty capsids in the nucleus of mutant-infected noncomplementing cells. We conclude that the ICP18.5(PrV) protein is necessary for viral replication and plays an essential role in the process of mature capsid formation.

Full text

PDF
1236

Images in this article

1239Image
on p.1239
1240Image
on p.1240
1240Image
on p.1240
1241Image
on p.1241
1242Image
on p.1242
1243Image
on p.1243

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Addison C., Rixon F. J., Preston V. G. Herpes simplex virus type 1 UL28 gene product is important for the formation of mature capsids. J Gen Virol. 1990 Oct;71(Pt 10):2377–2384. doi: 10.1099/0022-1317-71-10-2377. [DOI] [PubMed] [Google Scholar]
  2. Albrecht J. C., Fleckenstein B. Structural organization of the conserved gene block of Herpesvirus saimiri coding for DNA polymerase, glycoprotein B, and major DNA binding protein. Virology. 1990 Feb;174(2):533–542. doi: 10.1016/0042-6822(90)90107-3. [DOI] [PubMed] [Google Scholar]
  3. Baer R., Bankier A. T., Biggin M. D., Deininger P. L., Farrell P. J., Gibson T. J., Hatfull G., Hudson G. S., Satchwell S. C., Séguin C. DNA sequence and expression of the B95-8 Epstein-Barr virus genome. Nature. 1984 Jul 19;310(5974):207–211. doi: 10.1038/310207a0. [DOI] [PubMed] [Google Scholar]
  4. Bush M., Yager D. R., Gao M., Weisshart K., Marcy A. I., Coen D. M., Knipe D. M. Correct intranuclear localization of herpes simplex virus DNA polymerase requires the viral ICP8 DNA-binding protein. J Virol. 1991 Mar;65(3):1082–1089. doi: 10.1128/jvi.65.3.1082-1089.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bzik D. J., Debroy C., Fox B. A., Pederson N. E., Person S. The nucleotide sequence of the gB glycoprotein gene of HSV-2 and comparison with the corresponding gene of HSV-1. Virology. 1986 Dec;155(2):322–333. doi: 10.1016/0042-6822(86)90196-0. [DOI] [PubMed] [Google Scholar]
  6. Cai W. H., Gu B., Person S. Role of glycoprotein B of herpes simplex virus type 1 in viral entry and cell fusion. J Virol. 1988 Aug;62(8):2596–2604. doi: 10.1128/jvi.62.8.2596-2604.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chartrand P., Crumpacker C. S., Schaffer P. A., Wilkie N. M. Physical and genetic analysis of the herpes simplex virus DNA polymerase locus. Virology. 1980 Jun;103(2):311–326. doi: 10.1016/0042-6822(80)90190-7. [DOI] [PubMed] [Google Scholar]
  8. Chee M. S., Bankier A. T., Beck S., Bohni R., Brown C. M., Cerny R., Horsnell T., Hutchison C. A., 3rd, Kouzarides T., Martignetti J. A. Analysis of the protein-coding content of the sequence of human cytomegalovirus strain AD169. Curr Top Microbiol Immunol. 1990;154:125–169. doi: 10.1007/978-3-642-74980-3_6. [DOI] [PubMed] [Google Scholar]
  9. Crute J. J., Lehman I. R. Herpes simplex-1 DNA polymerase. Identification of an intrinsic 5'----3' exonuclease with ribonuclease H activity. J Biol Chem. 1989 Nov 15;264(32):19266–19270. [PubMed] [Google Scholar]
  10. 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]
  11. Deiss L. P., Frenkel N. Herpes simplex virus amplicon: cleavage of concatemeric DNA is linked to packaging and involves amplification of the terminally reiterated a sequence. J Virol. 1986 Mar;57(3):933–941. doi: 10.1128/jvi.57.3.933-941.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gao M., Knipe D. M. Potential role for herpes simplex virus ICP8 DNA replication protein in stimulation of late gene expression. J Virol. 1991 May;65(5):2666–2675. doi: 10.1128/jvi.65.5.2666-2675.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gibson W., Roizman B. Proteins specified by herpes simplex virus. 8. Characterization and composition of multiple capsid forms of subtypes 1 and 2. J Virol. 1972 Nov;10(5):1044–1052. doi: 10.1128/jvi.10.5.1044-1052.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gompels U. A., Craxton M. A., Honess R. W. Conservation of gene organization in the lymphotropic herpesviruses herpesvirus Saimiri and Epstein-Barr virus. J Virol. 1988 Mar;62(3):757–767. doi: 10.1128/jvi.62.3.757-767.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Griffin A. M. The nucleotide sequence of the glycoprotein gB gene of infectious laryngotracheitis virus: analysis and evolutionary relationship to the homologous gene from other herpesviruses. J Gen Virol. 1991 Feb;72(Pt 2):393–398. doi: 10.1099/0022-1317-72-2-393. [DOI] [PubMed] [Google Scholar]
  17. Hammerschmidt W., Conraths F., Mankertz J., Pauli G., Ludwig H., Buhk H. J. Conservation of a gene cluster including glycoprotein B in bovine herpesvirus type 2 (BHV-2) and herpes simplex virus type 1 (HSV-1). Virology. 1988 Aug;165(2):388–405. doi: 10.1016/0042-6822(88)90583-1. [DOI] [PubMed] [Google Scholar]
  18. Hampl H., Ben-Porat T., Ehrlicher L., Habermehl K. O., Kaplan A. S. Characterization of the envelope proteins of pseudorabies virus. J Virol. 1984 Nov;52(2):583–590. doi: 10.1128/jvi.52.2.583-590.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Holland L. E., Sandri-Goldin R. M., Goldin A. L., Glorioso J. C., Levine M. Transcriptional and genetic analyses of the herpes simplex virus type 1 genome: coordinates 0.29 to 0.45. J Virol. 1984 Mar;49(3):947–959. doi: 10.1128/jvi.49.3.947-959.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jonjić S., Koszinowski U. H. Monoclonal antibodies reactive with swine lymphocytes. I. Antibodies to membrane structures that define the cytolytic T lymphocyte subset in the swine. J Immunol. 1984 Aug;133(2):647–652. [PubMed] [Google Scholar]
  21. 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]
  22. Knopf C. W. The nucleotide sequence of the herpes simplex virus type 1 late gene ICP18.5 of strain Angelotti. Nucleic Acids Res. 1987 Oct 12;15(19):8109–8110. doi: 10.1093/nar/15.19.8109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ladin B. F., Blankenship M. L., Ben-Porat T. Replication of herpesvirus DNA. V. Maturation of concatemeric DNA of pseudorabies virus to genome length is related to capsid formation. J Virol. 1980 Mar;33(3):1151–1164. doi: 10.1128/jvi.33.3.1151-1164.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. McGeoch D. J., Dalrymple M. A., Davison A. J., Dolan A., Frame M. C., McNab D., Perry L. J., Scott J. E., Taylor P. The complete DNA sequence of the long unique region in the genome of herpes simplex virus type 1. J Gen Virol. 1988 Jul;69(Pt 7):1531–1574. doi: 10.1099/0022-1317-69-7-1531. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Orberg P. K., Schaffer P. A. Expression of herpes simplex virus type 1 major DNA-binding protein, ICP8, in transformed cell lines: complementation of deletion mutants and inhibition of wild-type virus. J Virol. 1987 Apr;61(4):1136–1146. doi: 10.1128/jvi.61.4.1136-1146.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pancake B. A., Aschman D. P., Schaffer P. A. Genetic and phenotypic analysis of herpes simplex virus type 1 mutants conditionally resistant to immune cytolysis. J Virol. 1983 Sep;47(3):568–585. doi: 10.1128/jvi.47.3.568-585.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Pederson N. E., Enquist L. W. Overexpression in bacterial and identification in infected cells of the pseudorabies virus protein homologous to herpes simplex virus type 1 ICP18.5. J Virol. 1991 Jul;65(7):3746–3758. doi: 10.1128/jvi.65.7.3746-3758.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pederson N. E., Enquist L. W. The nucleotide sequence of a pseudorabies virus gene similar to ICP18.5 of herpes simplex virus type 1. Nucleic Acids Res. 1989 May 11;17(9):3597–3597. doi: 10.1093/nar/17.9.3597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pellett P. E., Jenkins F. J., Ackermann M., Sarmiento M., Roizman B. Transcription initiation sites and nucleotide sequence of a herpes simplex virus 1 gene conserved in the Epstein-Barr virus genome and reported to affect the transport of viral glycoproteins. J Virol. 1986 Dec;60(3):1134–1140. doi: 10.1128/jvi.60.3.1134-1140.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. 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]
  34. Riggio M. P., Cullinane A. A., Onions D. E. Identification and nucleotide sequence of the glycoprotein gB gene of equine herpesvirus 4. J Virol. 1989 Mar;63(3):1123–1133. doi: 10.1128/jvi.63.3.1123-1133.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Robbins A. K., Dorney D. J., Wathen M. W., Whealy M. E., Gold C., Watson R. J., Holland L. E., Weed S. D., Levine M., Glorioso J. C. The pseudorabies virus gII gene is closely related to the gB glycoprotein gene of herpes simplex virus. J Virol. 1987 Sep;61(9):2691–2701. doi: 10.1128/jvi.61.9.2691-2701.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Robbins A. K., Watson R. J., Whealy M. E., Hays W. W., Enquist L. W. Characterization of a pseudorabies virus glycoprotein gene with homology to herpes simplex virus type 1 and type 2 glycoprotein C. J Virol. 1986 May;58(2):339–347. doi: 10.1128/jvi.58.2.339-347.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ross L. J., Sanderson M., Scott S. D., Binns M. M., Doel T., Milne B. Nucleotide sequence and characterization of the Marek's disease virus homologue of glycoprotein B of herpes simplex virus. J Gen Virol. 1989 Jul;70(Pt 7):1789–1804. doi: 10.1099/0022-1317-70-7-1789. [DOI] [PubMed] [Google Scholar]
  38. Rziha H. J., Mettenleiter T. C., Ohlinger V., Wittmann G. Herpesvirus (pseudorabies virus) latency in swine: occurrence and physical state of viral DNA in neural tissues. Virology. 1986 Dec;155(2):600–613. doi: 10.1016/0042-6822(86)90220-5. [DOI] [PubMed] [Google Scholar]
  39. Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
  40. Wang Y. S., Hall J. D. Characterization of a major DNA-binding domain in the herpes simplex virus type 1 DNA-binding protein (ICP8). J Virol. 1990 May;64(5):2082–2089. doi: 10.1128/jvi.64.5.2082-2089.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wathen M. W., Wathen L. M. Isolation, characterization, and physical mapping of a pseudorabies virus mutant containing antigenically altered gp50. J Virol. 1984 Jul;51(1):57–62. doi: 10.1128/jvi.51.1.57-62.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Weiland F., Keil G. M., Reddehase M. J., Koszinowski U. H. Studies on the morphogenesis of murine cytomegalovirus. Intervirology. 1986;26(4):192–201. doi: 10.1159/000149701. [DOI] [PubMed] [Google Scholar]
  43. Whalley J. M., Robertson G. R., Scott N. A., Hudson G. C., Bell C. W., Woodworth L. M. Identification and nucleotide sequence of a gene in equine herpesvirus 1 analogous to the herpes simplex virus gene encoding the major envelope glycoprotein gB. J Gen Virol. 1989 Feb;70(Pt 2):383–394. doi: 10.1099/0022-1317-70-2-383. [DOI] [PubMed] [Google Scholar]
  44. Whitbeck J. C., Bello L. J., Lawrence W. C. Comparison of the bovine herpesvirus 1 gI gene and the herpes simplex virus type 1 gB gene. J Virol. 1988 Sep;62(9):3319–3327. doi: 10.1128/jvi.62.9.3319-3327.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Wu C. A., Harper L., Ben-Porat T. cis Functions involved in replication and cleavage-encapsidation of pseudorabies virus. J Virol. 1986 Aug;59(2):318–327. doi: 10.1128/jvi.59.2.318-327.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES