Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1997 Jul;71(7):5639–5646. doi: 10.1128/jvi.71.7.5639-5646.1997

The UL20 gene product of pseudorabies virus functions in virus egress.

W Fuchs 1, B G Klupp 1, H Granzow 1, T C Mettenleiter 1
PMCID: PMC191809  PMID: 9188641

Abstract

The UL20 open reading frame is positionally conserved in different alphaherpesvirus genomes and is predicted to encode an integral membrane protein. A previously described UL20- mutant of herpes simplex virus type 1 (HSV-1) exhibited a defect in egress correlating with retention of virions in the perinuclear space (J. D. Baines, P. L. Ward, G. Campadelli-Fiume, and B. Roizman, J. Virol. 65:6414-6424, 1991). To analyze UL20 function in a related but different herpesvirus, we constructed a UL20- pseudorabies virus (PrV) mutant by insertional mutagenesis. Similar to HSV-1, UL20- PrV was found to be severely impaired in both cell-to-cell spread and release from cultured cells. The severity of this defect appeared to be cell type dependent, being more prominent in Vero than in human 143TK- cells. Surprisingly, electron microscopy revealed the retention of enveloped virus particles in cytoplasmic vesicles of Vero cells infected with UL20- PrV. This contrasts with the situation in the UL20- HSV-1 mutant, which accumulated virions in the perinuclear cisterna of Vero cells. Therefore, the UL20 gene products of PrV and HSV-1 appear to be involved in distinct steps of viral egress, acting in different intracellular compartments. This might be caused either by different functions of the UL20 proteins themselves or by generally different egress pathways of PrV and HSV-1 mediated by other viral gene products.

Full Text

The Full Text of this article is available as a PDF (1.6 MB).

Selected References

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

  1. Avitabile E., Ward P. L., Di Lazzaro C., Torrisi M. R., Roizman B., Campadelli-Fiume G. The herpes simplex virus UL20 protein compensates for the differential disruption of exocytosis of virions and viral membrane glycoproteins associated with fragmentation of the Golgi apparatus. J Virol. 1994 Nov;68(11):7397–7405. doi: 10.1128/jvi.68.11.7397-7405.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Babic N., Mettenleiter T. C., Ugolini G., Flamand A., Coulon P. Propagation of pseudorabies virus in the nervous system of the mouse after intranasal inoculation. Virology. 1994 Nov 1;204(2):616–625. doi: 10.1006/viro.1994.1576. [DOI] [PubMed] [Google Scholar]
  3. Baines J. D., Koyama A. H., Huang T., Roizman B. The UL21 gene products of herpes simplex virus 1 are dispensable for growth in cultured cells. J Virol. 1994 May;68(5):2929–2936. doi: 10.1128/jvi.68.5.2929-2936.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baines J. D., Ward P. L., Campadelli-Fiume G., Roizman B. The UL20 gene of herpes simplex virus 1 encodes a function necessary for viral egress. J Virol. 1991 Dec;65(12):6414–6424. doi: 10.1128/jvi.65.12.6414-6424.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Baumeister J., Klupp B. G., Mettenleiter T. C. Pseudorabies virus and equine herpesvirus 1 share a nonessential gene which is absent in other herpesviruses and located adjacent to a highly conserved gene cluster. J Virol. 1995 Sep;69(9):5560–5567. doi: 10.1128/jvi.69.9.5560-5567.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ben-Porat T., Veach R. A., Ihara S. Localization of the regions of homology between the genomes of herpes simplex virus, type 1, and pseudorabies virus. Virology. 1983 May;127(1):194–204. doi: 10.1016/0042-6822(83)90383-5. [DOI] [PubMed] [Google Scholar]
  7. Campadelli-Fiume G., Farabegoli F., Di Gaeta S., Roizman B. Origin of unenveloped capsids in the cytoplasm of cells infected with herpes simplex virus 1. J Virol. 1991 Mar;65(3):1589–1595. doi: 10.1128/jvi.65.3.1589-1595.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Campadelli G., Brandimarti R., Di Lazzaro C., Ward P. L., Roizman B., Torrisi M. R. Fragmentation and dispersal of Golgi proteins and redistribution of glycoproteins and glycolipids processed through the Golgi apparatus after infection with herpes simplex virus 1. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2798–2802. doi: 10.1073/pnas.90.7.2798. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Dean H. J., Cheung A. K. A 3' coterminal gene cluster in pseudorabies virus contains herpes simplex virus UL1, UL2, and UL3 gene homologs and a unique UL3.5 open reading frame. J Virol. 1993 Oct;67(10):5955–5961. doi: 10.1128/jvi.67.10.5955-5961.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dezélée S., Bras F., Vende P., Simonet B., Nguyen X., Flamand A., Masse M. J. The BamHI fragment 9 of pseudorabies virus contains genes homologous to the UL24, UL25, UL26, and UL 26.5 genes of herpes simplex virus type 1. Virus Res. 1996 Jun;42(1-2):27–39. doi: 10.1016/0168-1702(96)01293-2. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Gershon A. A., Sherman D. L., Zhu Z., Gabel C. A., Ambron R. T., Gershon M. D. Intracellular transport of newly synthesized varicella-zoster virus: final envelopment in the trans-Golgi network. J Virol. 1994 Oct;68(10):6372–6390. doi: 10.1128/jvi.68.10.6372-6390.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Granzow H., Weiland F., Jöns A., Klupp B. G., Karger A., Mettenleiter T. C. Ultrastructural analysis of the replication cycle of pseudorabies virus in cell culture: a reassessment. J Virol. 1997 Mar;71(3):2072–2082. doi: 10.1128/jvi.71.3.2072-2082.1997. [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. Klupp B. G., Kern H., Mettenleiter T. C. The virulence-determining genomic BamHI fragment 4 of pseudorabies virus contains genes corresponding to the UL15 (partial), UL18, UL19, UL20, and UL21 genes of herpes simplex virus and a putative origin of replication. Virology. 1992 Dec;191(2):900–908. doi: 10.1016/0042-6822(92)90265-q. [DOI] [PubMed] [Google Scholar]
  18. Klupp B. G., Lomniczi B., Visser N., Fuchs W., Mettenleiter T. C. Mutations affecting the UL21 gene contribute to avirulence of pseudorabies virus vaccine strain Bartha. Virology. 1995 Oct 1;212(2):466–473. doi: 10.1006/viro.1995.1504. [DOI] [PubMed] [Google Scholar]
  19. Klupp B. G., Mettenleiter T. C. Sequence and expression of the glycoprotein gH gene of pseudorabies virus. Virology. 1991 Jun;182(2):732–741. doi: 10.1016/0042-6822(91)90614-h. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. MacLean C. A., Efstathiou S., Elliott M. L., Jamieson F. E., McGeoch D. J. Investigation of herpes simplex virus type 1 genes encoding multiply inserted membrane proteins. J Gen Virol. 1991 Apr;72(Pt 4):897–906. doi: 10.1099/0022-1317-72-4-897. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Mettenleiter T. C., Kern H., Rauh I. Isolation of a viable herpesvirus (pseudorabies virus) mutant specifically lacking all four known nonessential glycoproteins. Virology. 1990 Nov;179(1):498–503. doi: 10.1016/0042-6822(90)90324-k. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. Roizmann B., Desrosiers R. C., Fleckenstein B., Lopez C., Minson A. C., Studdert M. J. The family Herpesviridae: an update. The Herpesvirus Study Group of the International Committee on Taxonomy of Viruses. Arch Virol. 1992;123(3-4):425–449. doi: 10.1007/BF01317276. [DOI] [PubMed] [Google Scholar]
  27. Sanes J. R., Rubenstein J. L., Nicolas J. F. Use of a recombinant retrovirus to study post-implantation cell lineage in mouse embryos. EMBO J. 1986 Dec 1;5(12):3133–3142. doi: 10.1002/j.1460-2075.1986.tb04620.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schwyzer M., Ackermann M. Molecular virology of ruminant herpesviruses. Vet Microbiol. 1996 Nov;53(1-2):17–29. doi: 10.1016/s0378-1135(96)01231-x. [DOI] [PubMed] [Google Scholar]
  29. Telford E. A., Watson M. S., McBride K., Davison A. J. The DNA sequence of equine herpesvirus-1. Virology. 1992 Jul;189(1):304–316. doi: 10.1016/0042-6822(92)90706-u. [DOI] [PubMed] [Google Scholar]
  30. Torrisi M. R., Di Lazzaro C., Pavan A., Pereira L., Campadelli-Fiume G. Herpes simplex virus envelopment and maturation studied by fracture label. J Virol. 1992 Jan;66(1):554–561. doi: 10.1128/jvi.66.1.554-561.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Vlcek C., Benes V., Lu Z., Kutish G. F., Paces V., Rock D., Letchworth G. J., Schwyzer M. Nucleotide sequence analysis of a 30-kb region of the bovine herpesvirus 1 genome which exhibits a colinear gene arrangement with the UL21 to UL4 genes of herpes simplex virus. Virology. 1995 Jun 20;210(1):100–108. doi: 10.1006/viro.1995.1321. [DOI] [PubMed] [Google Scholar]
  32. Ward P. L., Campadelli-Fiume G., Avitabile E., Roizman B. Localization and putative function of the UL20 membrane protein in cells infected with herpes simplex virus 1. J Virol. 1994 Nov;68(11):7406–7417. doi: 10.1128/jvi.68.11.7406-7417.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Whealy M. E., Card J. P., Meade R. P., Robbins A. K., Enquist L. W. Effect of brefeldin A on alphaherpesvirus membrane protein glycosylation and virus egress. J Virol. 1991 Mar;65(3):1066–1081. doi: 10.1128/jvi.65.3.1066-1081.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Whealy M. E., Robbins A. K., Tufaro F., Enquist L. W. A cellular function is required for pseudorabies virus envelope glycoprotein processing and virus egress. J Virol. 1992 Jun;66(6):3803–3810. doi: 10.1128/jvi.66.6.3803-3810.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. de Wind N., Wagenaar F., Pol J., Kimman T., Berns A. The pseudorabies virus homology of the herpes simplex virus UL21 gene product is a capsid protein which is involved in capsid maturation. J Virol. 1992 Dec;66(12):7096–7103. doi: 10.1128/jvi.66.12.7096-7103.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES