Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1996 Dec;70(12):8402–8410. doi: 10.1128/jvi.70.12.8402-8410.1996

Accessory human cytomegalovirus glycoprotein US9 in the unique short component of the viral genome promotes cell-to-cell transmission of virus in polarized epithelial cells.

E Maidji 1, S Tugizov 1, T Jones 1, Z Zheng 1, L Pereira 1
PMCID: PMC190929  PMID: 8970961

Abstract

Human cytomegalovirus (CMV) encodes accessory glycoproteins that are dispensable for virus growth in nonpolarized cells in culture. We report that CMV deletion mutants lacking the gene for accessory glycoprotein US9 in the unique short component of the viral genome are impaired in plaque formation in polarized human retinal pigment epithelial (ARPE-19) cells. Comparison of CMV deletion mutants in US9 with herpes simplex virus type 1 deletion mutants lacking glycoproteins gE and gI showed that both of these mutants are impaired in altering junctional complexes and increasing paracellular permeability in polarized ARPE-19 cells cultured on permeable filter supports. Results of functional studies indicate that CMV US9 and homologs of gE have analogous roles in promoting virus spread across lateral membranes of polarized epithelial cells.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. 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]
  2. Baines J. D., Roizman B. The UL10 gene of herpes simplex virus 1 encodes a novel viral glycoprotein, gM, which is present in the virion and in the plasma membrane of infected cells. J Virol. 1993 Mar;67(3):1441–1452. doi: 10.1128/jvi.67.3.1441-1452.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Balan P., Davis-Poynter N., Bell S., Atkinson H., Browne H., Minson T. An analysis of the in vitro and in vivo phenotypes of mutants of herpes simplex virus type 1 lacking glycoproteins gG, gE, gI or the putative gJ. J Gen Virol. 1994 Jun;75(Pt 6):1245–1258. doi: 10.1099/0022-1317-75-6-1245. [DOI] [PubMed] [Google Scholar]
  4. Baudry B., Fasano A., Ketley J., Kaper J. B. Cloning of a gene (zot) encoding a new toxin produced by Vibrio cholerae. Infect Immun. 1992 Feb;60(2):428–434. doi: 10.1128/iai.60.2.428-434.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Card J. P., Whealy M. E., Robbins A. K., Enquist L. W. Pseudorabies virus envelope glycoprotein gI influences both neurotropism and virulence during infection of the rat visual system. J Virol. 1992 May;66(5):3032–3041. doi: 10.1128/jvi.66.5.3032-3041.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Card J. P., Whealy M. E., Robbins A. K., Moore R. Y., Enquist L. W. Two alpha-herpesvirus strains are transported differentially in the rodent visual system. Neuron. 1991 Jun;6(6):957–969. doi: 10.1016/0896-6273(91)90236-s. [DOI] [PubMed] [Google Scholar]
  7. Chakraborty T., Ebel F., Domann E., Niebuhr K., Gerstel B., Pistor S., Temm-Grove C. J., Jockusch B. M., Reinhard M., Walter U. A focal adhesion factor directly linking intracellularly motile Listeria monocytogenes and Listeria ivanovii to the actin-based cytoskeleton of mammalian cells. EMBO J. 1995 Apr 3;14(7):1314–1321. doi: 10.1002/j.1460-2075.1995.tb07117.x. [DOI] [PMC free article] [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. Cossart P. Actin-based bacterial motility. Curr Opin Cell Biol. 1995 Feb;7(1):94–101. doi: 10.1016/0955-0674(95)80050-6. [DOI] [PubMed] [Google Scholar]
  10. Coster T. S., Killeen K. P., Waldor M. K., Beattie D. T., Spriggs D. R., Kenner J. R., Trofa A., Sadoff J. C., Mekalanos J. J., Taylor D. N. Safety, immunogenicity, and efficacy of live attenuated Vibrio cholerae O139 vaccine prototype. Lancet. 1995 Apr 15;345(8955):949–952. doi: 10.1016/s0140-6736(95)90698-3. [DOI] [PubMed] [Google Scholar]
  11. Cudmore S., Cossart P., Griffiths G., Way M. Actin-based motility of vaccinia virus. Nature. 1995 Dec 7;378(6557):636–638. doi: 10.1038/378636a0. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Detrick B., Rhame J., Wang Y., Nagineni C. N., Hooks J. J. Cytomegalovirus replication in human retinal pigment epithelial cells. Altered expression of viral early proteins. Invest Ophthalmol Vis Sci. 1996 Apr;37(5):814–825. [PubMed] [Google Scholar]
  14. Dingwell K. S., Brunetti C. R., Hendricks R. L., Tang Q., Tang M., Rainbow A. J., Johnson D. C. Herpes simplex virus glycoproteins E and I facilitate cell-to-cell spread in vivo and across junctions of cultured cells. J Virol. 1994 Feb;68(2):834–845. doi: 10.1128/jvi.68.2.834-845.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Dingwell K. S., Doering L. C., Johnson D. C. Glycoproteins E and I facilitate neuron-to-neuron spread of herpes simplex virus. J Virol. 1995 Nov;69(11):7087–7098. doi: 10.1128/jvi.69.11.7087-7098.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Drew W. L. Cytomegalovirus infection in patients with AIDS. J Infect Dis. 1988 Aug;158(2):449–456. doi: 10.1093/infdis/158.2.449. [DOI] [PubMed] [Google Scholar]
  17. Dunn K. C., Aotaki-Keen A. E., Putkey F. R., Hjelmeland L. M. ARPE-19, a human retinal pigment epithelial cell line with differentiated properties. Exp Eye Res. 1996 Feb;62(2):155–169. doi: 10.1006/exer.1996.0020. [DOI] [PubMed] [Google Scholar]
  18. Elton D. M., Halliburton I. W., Killington R. A., Meredith D. M., Bonass W. A. Sequence analysis of the 4.7-kb BamHI-EcoRI fragment of the equine herpesvirus type-1 short unique region. Gene. 1991 May 30;101(2):203–208. doi: 10.1016/0378-1119(91)90412-5. [DOI] [PubMed] [Google Scholar]
  19. Enquist L. W., Dubin J., Whealy M. E., Card J. P. Complementation analysis of pseudorabies virus gE and gI mutants in retinal ganglion cell neurotropism. J Virol. 1994 Aug;68(8):5275–5279. doi: 10.1128/jvi.68.8.5275-5279.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Fasano A., Baudry B., Pumplin D. W., Wasserman S. S., Tall B. D., Ketley J. M., Kaper J. B. Vibrio cholerae produces a second enterotoxin, which affects intestinal tight junctions. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5242–5246. doi: 10.1073/pnas.88.12.5242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Fasano A., Fiorentini C., Donelli G., Uzzau S., Kaper J. B., Margaretten K., Ding X., Guandalini S., Comstock L., Goldblum S. E. Zonula occludens toxin modulates tight junctions through protein kinase C-dependent actin reorganization, in vitro. J Clin Invest. 1995 Aug;96(2):710–720. doi: 10.1172/JCI118114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Fath K. R., Burgess D. R. Golgi-derived vesicles from developing epithelial cells bind actin filaments and possess myosin-I as a cytoplasmically oriented peripheral membrane protein. J Cell Biol. 1993 Jan;120(1):117–127. doi: 10.1083/jcb.120.1.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Gompels U. A., Nicholas J., Lawrence G., Jones M., Thomson B. J., Martin M. E., Efstathiou S., Craxton M., Macaulay H. A. The DNA sequence of human herpesvirus-6: structure, coding content, and genome evolution. Virology. 1995 May 10;209(1):29–51. doi: 10.1006/viro.1995.1228. [DOI] [PubMed] [Google Scholar]
  24. Gumbiner B. Structure, biochemistry, and assembly of epithelial tight junctions. Am J Physiol. 1987 Dec;253(6 Pt 1):C749–C758. doi: 10.1152/ajpcell.1987.253.6.C749. [DOI] [PubMed] [Google Scholar]
  25. Johnson D. C., Frame M. C., Ligas M. W., Cross A. M., Stow N. D. Herpes simplex virus immunoglobulin G Fc receptor activity depends on a complex of two viral glycoproteins, gE and gI. J Virol. 1988 Apr;62(4):1347–1354. doi: 10.1128/jvi.62.4.1347-1354.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Johnson J. A., Morris J. G., Jr, Kaper J. B. Gene encoding zonula occludens toxin (zot) does not occur independently from cholera enterotoxin genes (ctx) in Vibrio cholerae. J Clin Microbiol. 1993 Mar;31(3):732–733. doi: 10.1128/jcm.31.3.732-733.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Jones N. L., Lewis J. C., Kilpatrick B. A. Cytoskeletal disruption during human cytomegalovirus infection of human lung fibroblasts. Eur J Cell Biol. 1986 Aug;41(2):304–312. [PubMed] [Google Scholar]
  28. Jones T. R., Hanson L. K., Sun L., Slater J. S., Stenberg R. M., Campbell A. E. Multiple independent loci within the human cytomegalovirus unique short region down-regulate expression of major histocompatibility complex class I heavy chains. J Virol. 1995 Aug;69(8):4830–4841. doi: 10.1128/jvi.69.8.4830-4841.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Jones T. R., Muzithras V. P. A cluster of dispensable genes within the human cytomegalovirus genome short component: IRS1, US1 through US5, and the US6 family. J Virol. 1992 Apr;66(4):2541–2546. doi: 10.1128/jvi.66.4.2541-2546.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Jones T. R., Muzithras V. P. Fine mapping of transcripts expressed from the US6 gene family of human cytomegalovirus strain AD169. J Virol. 1991 Apr;65(4):2024–2036. doi: 10.1128/jvi.65.4.2024-2036.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Jones T. R., Muzithras V. P., Gluzman Y. Replacement mutagenesis of the human cytomegalovirus genome: US10 and US11 gene products are nonessential. J Virol. 1991 Nov;65(11):5860–5872. doi: 10.1128/jvi.65.11.5860-5872.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Kemble G. W., McCormick A. L., Pereira L., Mocarski E. S. A cytomegalovirus protein with properties of herpes simplex virus ICP8: partial purification of the polypeptide and map position of the gene. J Virol. 1987 Oct;61(10):3143–3151. doi: 10.1128/jvi.61.10.3143-3151.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Kollert-Jöns A., Bogner E., Radsak K. A 15-kilobase-pair region of the human cytomegalovirus genome which includes US1 through US13 is dispensable for growth in cell culture. J Virol. 1991 Oct;65(10):5184–5189. doi: 10.1128/jvi.65.10.5184-5189.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Kuznetsov S. A., Langford G. M., Weiss D. G. Actin-dependent organelle movement in squid axoplasm. Nature. 1992 Apr 23;356(6371):722–725. doi: 10.1038/356722a0. [DOI] [PubMed] [Google Scholar]
  35. Leung-Tack P., Audonnet J. C., Riviere M. The complete DNA sequence and the genetic organization of the short unique region (US) of the bovine herpesvirus type 1 (ST strain). Virology. 1994 Mar;199(2):409–421. doi: 10.1006/viro.1994.1139. [DOI] [PubMed] [Google Scholar]
  36. Litwin V., Jackson W., Grose C. Receptor properties of two varicella-zoster virus glycoproteins, gpI and gpIV, homologous to herpes simplex virus gE and gI. J Virol. 1992 Jun;66(6):3643–3651. doi: 10.1128/jvi.66.6.3643-3651.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Longnecker R., Chatterjee S., Whitley R. J., Roizman B. Identification of a herpes simplex virus 1 glycoprotein gene within a gene cluster dispensable for growth in cell culture. Proc Natl Acad Sci U S A. 1987 Jun;84(12):4303–4307. doi: 10.1073/pnas.84.12.4303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Longnecker R., Roizman B. Clustering of genes dispensable for growth in culture in the S component of the HSV-1 genome. Science. 1987 May 1;236(4801):573–576. doi: 10.1126/science.3033823. [DOI] [PubMed] [Google Scholar]
  39. Madara J. L., Barenberg D., Carlson S. Effects of cytochalasin D on occluding junctions of intestinal absorptive cells: further evidence that the cytoskeleton may influence paracellular permeability and junctional charge selectivity. J Cell Biol. 1986 Jun;102(6):2125–2136. doi: 10.1083/jcb.102.6.2125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Mays R. W., Beck K. A., Nelson W. J. Organization and function of the cytoskeleton in polarized epithelial cells: a component of the protein sorting machinery. Curr Opin Cell Biol. 1994 Feb;6(1):16–24. doi: 10.1016/0955-0674(94)90111-2. [DOI] [PubMed] [Google Scholar]
  41. McGeoch D. J., Dolan A., Donald S., Rixon F. J. Sequence determination and genetic content of the short unique region in the genome of herpes simplex virus type 1. J Mol Biol. 1985 Jan 5;181(1):1–13. doi: 10.1016/0022-2836(85)90320-1. [DOI] [PubMed] [Google Scholar]
  42. Mettenleiter T. C., Zsak L., Kaplan A. S., Ben-Porat T., Lomniczi B. Role of a structural glycoprotein of pseudorabies in virus virulence. J Virol. 1987 Dec;61(12):4030–4032. doi: 10.1128/jvi.61.12.4030-4032.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Miceli M. V., Newsome D. A., Novak L. C., Beuerman R. W. Cytomegalovirus replication in cultured human retinal pigment epithelial cells. Curr Eye Res. 1989 Aug;8(8):835–839. doi: 10.3109/02713688909000873. [DOI] [PubMed] [Google Scholar]
  44. Mocarski E. S., Pereira L., Michael N. Precise localization of genes on large animal virus genomes: use of lambda gt11 and monoclonal antibodies to map the gene for a cytomegalovirus protein family. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1266–1270. doi: 10.1073/pnas.82.4.1266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Navarro D., Paz P., Pereira L. Domains of herpes simplex virus I glycoprotein B that function in virus penetration, cell-to-cell spread, and cell fusion. Virology. 1992 Jan;186(1):99–112. doi: 10.1016/0042-6822(92)90064-v. [DOI] [PubMed] [Google Scholar]
  46. Navarro D., Paz P., Tugizov S., Topp K., La Vail J., Pereira L. Glycoprotein B of human cytomegalovirus promotes virion penetration into cells, transmission of infection from cell to cell, and fusion of infected cells. Virology. 1993 Nov;197(1):143–158. doi: 10.1006/viro.1993.1575. [DOI] [PubMed] [Google Scholar]
  47. Näthke I. S., Hinck L. E., Nelson W. J. Epithelial cell adhesion and development of cell surface polarity: possible mechanisms for modulation of cadherin function, organization and distribution. J Cell Sci Suppl. 1993;17:139–145. doi: 10.1242/jcs.1993.supplement_17.20. [DOI] [PubMed] [Google Scholar]
  48. Pereira L. Function of glycoprotein B homologues of the family herpesviridae. Infect Agents Dis. 1994 Feb;3(1):9–28. [PubMed] [Google Scholar]
  49. Pereira L., Hoffman M., Gallo D., Cremer N. Monoclonal antibodies to human cytomegalovirus: three surface membrane proteins with unique immunological and electrophoretic properties specify cross-reactive determinants. Infect Immun. 1982 Jun;36(3):924–932. doi: 10.1128/iai.36.3.924-932.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Pereira L., Maidji E., Tugizov S., Jones T. Deletion mutants in human cytomegalovirus glycoprotein US9 are impaired in cell-cell transmission and in altering tight junctions of polarized human retinal pigment epithelial cells. Scand J Infect Dis Suppl. 1995;99:82–87. [PubMed] [Google Scholar]
  51. Petrovskis E. A., Timmins J. G., Post L. E. Use of lambda gt11 to isolate genes for two pseudorabies virus glycoproteins with homology to herpes simplex virus and varicella-zoster virus glycoproteins. J Virol. 1986 Oct;60(1):185–193. doi: 10.1128/jvi.60.1.185-193.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Pollard T. D. Actin cytoskeleton. Missing link for intracellular bacterial motility? Curr Biol. 1995 Aug 1;5(8):837–840. doi: 10.1016/s0960-9822(95)00167-9. [DOI] [PubMed] [Google Scholar]
  53. Quint W., Gielkens A., Van Oirschot J., Berns A., Cuypers H. T. Construction and characterization of deletion mutants of pseudorabies virus: a new generation of 'live' vaccines. J Gen Virol. 1987 Feb;68(Pt 2):523–534. doi: 10.1099/0022-1317-68-2-523. [DOI] [PubMed] [Google Scholar]
  54. Rodriguez-Boulan E., Powell S. K. Polarity of epithelial and neuronal cells. Annu Rev Cell Biol. 1992;8:395–427. doi: 10.1146/annurev.cb.08.110192.002143. [DOI] [PubMed] [Google Scholar]
  55. 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]
  56. Sears A. E., McGwire B. S., Roizman B. Infection of polarized MDCK cells with herpes simplex virus 1: two asymmetrically distributed cell receptors interact with different viral proteins. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5087–5091. doi: 10.1073/pnas.88.12.5087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Stamey F. R., Dominguez G., Black J. B., Dambaugh T. R., Pellett P. E. Intragenomic linear amplification of human herpesvirus 6B oriLyt suggests acquisition of oriLyt by transposition. J Virol. 1995 Jan;69(1):589–596. doi: 10.1128/jvi.69.1.589-596.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Theriot J. A. Bacterial pathogens caught in the actin. Curr Biol. 1992 Dec;2(12):649–651. doi: 10.1016/0960-9822(92)90115-q. [DOI] [PubMed] [Google Scholar]
  59. Trucksis M., Galen J. E., Michalski J., Fasano A., Kaper J. B. Accessory cholera enterotoxin (Ace), the third toxin of a Vibrio cholerae virulence cassette. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5267–5271. doi: 10.1073/pnas.90.11.5267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Tugizov S., Maidji E., Pereira L. Role of apical and basolateral membranes in replication of human cytomegalovirus in polarized retinal pigment epithelial cells. J Gen Virol. 1996 Jan;77(Pt 1):61–74. doi: 10.1099/0022-1317-77-1-61. [DOI] [PubMed] [Google Scholar]
  61. Waldor M. K., Mekalanos J. J. Emergence of a new cholera pandemic: molecular analysis of virulence determinants in Vibrio cholerae O139 and development of a live vaccine prototype. J Infect Dis. 1994 Aug;170(2):278–283. doi: 10.1093/infdis/170.2.278. [DOI] [PubMed] [Google Scholar]
  62. Whealy M. E., Card J. P., Robbins A. K., Dubin J. R., Rziha H. J., Enquist L. W. Specific pseudorabies virus infection of the rat visual system requires both gI and gp63 glycoproteins. J Virol. 1993 Jul;67(7):3786–3797. doi: 10.1128/jvi.67.7.3786-3797.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Wiertz E. J., Jones T. R., Sun L., Bogyo M., Geuze H. J., Ploegh H. L. The human cytomegalovirus US11 gene product dislocates MHC class I heavy chains from the endoplasmic reticulum to the cytosol. Cell. 1996 Mar 8;84(5):769–779. doi: 10.1016/s0092-8674(00)81054-5. [DOI] [PubMed] [Google Scholar]
  64. Yamashita Y., Shimokata K., Saga S., Mizuno S., Tsurumi T., Nishiyama Y. Rapid degradation of the heavy chain of class I major histocompatibility complex antigens in the endoplasmic reticulum of human cytomegalovirus-infected cells. J Virol. 1994 Dec;68(12):7933–7943. doi: 10.1128/jvi.68.12.7933-7943.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Zuckermann F. A., Mettenleiter T. C., Schreurs C., Sugg N., Ben-Porat T. Complex between glycoproteins gI and gp63 of pseudorabies virus: its effect on virus replication. J Virol. 1988 Dec;62(12):4622–4626. doi: 10.1128/jvi.62.12.4622-4626.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES