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. 1992 Nov;66(11):6609–6615. doi: 10.1128/jvi.66.11.6609-6615.1992

The UL16 gene of human cytomegalovirus encodes a glycoprotein that is dispensable for growth in vitro.

J Kaye 1, H Browne 1, M Stoffel 1, T Minson 1
PMCID: PMC240156  PMID: 1328682

Abstract

The UL16 gene of human cytomegalovirus (HCMV) encodes a predicted translation product with features characteristic of glycoproteins (signal and anchor sequences and eight potential N-linked glycosylation sites). Antisera were raised against the UL16 gene product expressed in Escherichia coli as a beta-galactosidase fusion protein. The antisera detected a 50-kDa glycoprotein in HCMV-infected cells that was absent from purified virions. The UL16 glycoprotein was synthesized at early times after infection and accumulated to the highest levels at late times after infection. A recombinant HCMV in which UL16 coding sequences were interrupted by a lacZ expression cassette was constructed by insertional mutagenesis. Analysis of the phenotype of the recombinant virus indicated that the UL16 gene product is nonessential for virus infectivity and growth in tissue culture.

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Selected References

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  1. Beck S., Barrell B. G. Human cytomegalovirus encodes a glycoprotein homologous to MHC class-I antigens. Nature. 1988 Jan 21;331(6153):269–272. doi: 10.1038/331269a0. [DOI] [PubMed] [Google Scholar]
  2. Britt W. J., Auger D. Synthesis and processing of the envelope gp55-116 complex of human cytomegalovirus. J Virol. 1986 Apr;58(1):185–191. doi: 10.1128/jvi.58.1.185-191.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Britt W. J., Vugler L. G. Processing of the gp55-116 envelope glycoprotein complex (gB) of human cytomegalovirus. J Virol. 1989 Jan;63(1):403–410. doi: 10.1128/jvi.63.1.403-410.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Browne H., Churcher M., Minson T. Construction and characterization of a human cytomegalovirus mutant with the UL18 (class I homolog) gene deleted. J Virol. 1992 Nov;66(11):6784–6787. doi: 10.1128/jvi.66.11.6784-6787.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Browne H., Smith G., Beck S., Minson T. A complex between the MHC class I homologue encoded by human cytomegalovirus and beta 2 microglobulin. Nature. 1990 Oct 25;347(6295):770–772. doi: 10.1038/347770a0. [DOI] [PubMed] [Google Scholar]
  6. Bzik D. J., Fox B. A., DeLuca N. A., Person S. Nucleotide sequence specifying the glycoprotein gene, gB, of herpes simplex virus type 1. Virology. 1984 Mar;133(2):301–314. doi: 10.1016/0042-6822(84)90397-0. [DOI] [PubMed] [Google Scholar]
  7. 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]
  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. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cranage M. P., Kouzarides T., Bankier A. T., Satchwell S., Weston K., Tomlinson P., Barrell B., Hart H., Bell S. E., Minson A. C. Identification of the human cytomegalovirus glycoprotein B gene and induction of neutralizing antibodies via its expression in recombinant vaccinia virus. EMBO J. 1986 Nov;5(11):3057–3063. doi: 10.1002/j.1460-2075.1986.tb04606.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cranage M. P., Smith G. L., Bell S. E., Hart H., Brown C., Bankier A. T., Tomlinson P., Barrell B. G., Minson T. C. Identification and expression of a human cytomegalovirus glycoprotein with homology to the Epstein-Barr virus BXLF2 product, varicella-zoster virus gpIII, and herpes simplex virus type 1 glycoprotein H. J Virol. 1988 Apr;62(4):1416–1422. doi: 10.1128/jvi.62.4.1416-1422.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Davison A. J., Edson C. M., Ellis R. W., Forghani B., Gilden D., Grose C., Keller P. M., Vafai A., Wroblewska Z., Yamanishi K. New common nomenclature for glycoprotein genes of varicella-zoster virus and their glycosylated products. J Virol. 1986 Mar;57(3):1195–1197. doi: 10.1128/jvi.57.3.1195-1197.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Farrar G. H., Greenaway P. J. Characterization of glycoprotein complexes present in human cytomegalovirus envelopes. J Gen Virol. 1986 Jul;67(Pt 7):1469–1473. doi: 10.1099/0022-1317-67-7-1469. [DOI] [PubMed] [Google Scholar]
  14. Forrester A., Farrell H., Wilkinson G., Kaye J., Davis-Poynter N., Minson T. Construction and properties of a mutant of herpes simplex virus type 1 with glycoprotein H coding sequences deleted. J Virol. 1992 Jan;66(1):341–348. doi: 10.1128/jvi.66.1.341-348.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gompels U., Minson A. The properties and sequence of glycoprotein H of herpes simplex virus type 1. Virology. 1986 Sep;153(2):230–247. doi: 10.1016/0042-6822(86)90026-7. [DOI] [PubMed] [Google Scholar]
  16. Greenaway P. J., Wilkinson G. W. Nucleotide sequence of the most abundantly transcribed early gene of human cytomegalovirus strain AD169. Virus Res. 1987 Feb;7(1):17–31. doi: 10.1016/0168-1702(87)90055-4. [DOI] [PubMed] [Google Scholar]
  17. Gretch D. R., Gehrz R. C., Stinski M. F. Characterization of a human cytomegalovirus glycoprotein complex (gcI). J Gen Virol. 1988 Jun;69(Pt 6):1205–1215. doi: 10.1099/0022-1317-69-6-1205. [DOI] [PubMed] [Google Scholar]
  18. Gretch D. R., Kari B., Gehrz R. C., Stinski M. F. A multigene family encodes the human cytomegalovirus glycoprotein complex gcII (gp47-52 complex). J Virol. 1988 Jun;62(6):1956–1962. doi: 10.1128/jvi.62.6.1956-1962.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gretch D. R., Kari B., Rasmussen L., Gehrz R. C., Stinski M. F. Identification and characterization of three distinct families of glycoprotein complexes in the envelopes of human cytomegalovirus. J Virol. 1988 Mar;62(3):875–881. doi: 10.1128/jvi.62.3.875-881.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hutchinson L., Browne H., Wargent V., Davis-Poynter N., Primorac S., Goldsmith K., Minson A. C., Johnson D. C. A novel herpes simplex virus glycoprotein, gL, forms a complex with glycoprotein H (gH) and affects normal folding and surface expression of gH. J Virol. 1992 Apr;66(4):2240–2250. doi: 10.1128/jvi.66.4.2240-2250.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. 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]
  23. Kari B., Gehrz R. A human cytomegalovirus glycoprotein complex designated gC-II is a major heparin-binding component of the envelope. J Virol. 1992 Mar;66(3):1761–1764. doi: 10.1128/jvi.66.3.1761-1764.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kari B., Lussenhop N., Goertz R., Wabuke-Bunoti M., Radeke R., Gehrz R. Characterization of monoclonal antibodies reactive to several biochemically distinct human cytomegalovirus glycoprotein complexes. J Virol. 1986 Nov;60(2):345–352. doi: 10.1128/jvi.60.2.345-352.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Keller P. M., Davison A. J., Lowe R. S., Bennett C. D., Ellis R. W. Identification and structure of the gene encoding gpII, a major glycoprotein of varicella-zoster virus. Virology. 1986 Jul 15;152(1):181–191. doi: 10.1016/0042-6822(86)90383-1. [DOI] [PubMed] [Google Scholar]
  26. Mackett M., Smith G. L., Moss B. General method for production and selection of infectious vaccinia virus recombinants expressing foreign genes. J Virol. 1984 Mar;49(3):857–864. doi: 10.1128/jvi.49.3.857-864.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. McDonough S. H., Staprans S. I., Spector D. H. Analysis of the major transcripts encoded by the long repeat of human cytomegalovirus strain AD169. J Virol. 1985 Mar;53(3):711–718. doi: 10.1128/jvi.53.3.711-718.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. McGeoch D. J., Davison A. J. DNA sequence of the herpes simplex virus type 1 gene encoding glycoprotein gH, and identification of homologues in the genomes of varicella-zoster virus and Epstein-Barr virus. Nucleic Acids Res. 1986 May 27;14(10):4281–4292. doi: 10.1093/nar/14.10.4281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Oram J. D., Downing R. G., Akrigg A., Dollery A. A., Duggleby C. J., Wilkinson G. W., Greenaway P. J. Use of recombinant plasmids to investigate the structure of the human cytomegalovirus genome. J Gen Virol. 1982 Mar;59(Pt 1):111–129. doi: 10.1099/0022-1317-59-1-111. [DOI] [PubMed] [Google Scholar]
  30. Pachl C., Probert W. S., Hermsen K. M., Masiarz F. R., Rasmussen L., Merigan T. C., Spaete R. R. The human cytomegalovirus strain Towne glycoprotein H gene encodes glycoprotein p86. Virology. 1989 Apr;169(2):418–426. doi: 10.1016/0042-6822(89)90167-0. [DOI] [PubMed] [Google Scholar]
  31. Pellett P. E., Biggin M. D., Barrell B., Roizman B. Epstein-Barr virus genome may encode a protein showing significant amino acid and predicted secondary structure homology with glycoprotein B of herpes simplex virus 1. J Virol. 1985 Dec;56(3):807–813. doi: 10.1128/jvi.56.3.807-813.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Pellett P. E., Kousoulas K. G., Pereira L., Roizman B. Anatomy of the herpes simplex virus 1 strain F glycoprotein B gene: primary sequence and predicted protein structure of the wild type and of monoclonal antibody-resistant mutants. J Virol. 1985 Jan;53(1):243–253. doi: 10.1128/jvi.53.1.243-253.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Pereira L., Hoffman M., Tatsuno M., Dondero D. Polymorphism of human cytomegalovirus glycoproteins characterized by monoclonal antibodies. Virology. 1984 Nov;139(1):73–86. doi: 10.1016/0042-6822(84)90331-3. [DOI] [PubMed] [Google Scholar]
  35. Rasmussen L. E., Nelson R. M., Kelsall D. C., Merigan T. C. Murine monoclonal antibody to a single protein neutralizes the infectivity of human cytomegalovirus. Proc Natl Acad Sci U S A. 1984 Feb;81(3):876–880. doi: 10.1073/pnas.81.3.876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rasmussen L., Mullenax J., Nelson R., Merigan T. C. Viral polypeptides detected by a complement-dependent neutralizing murine monoclonal antibody to human cytomegalovirus. J Virol. 1985 Aug;55(2):274–280. doi: 10.1128/jvi.55.2.274-280.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Spaete R. R., Mocarski E. S. Insertion and deletion mutagenesis of the human cytomegalovirus genome. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7213–7217. doi: 10.1073/pnas.84.20.7213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Stanley K. K., Luzio J. P. Construction of a new family of high efficiency bacterial expression vectors: identification of cDNA clones coding for human liver proteins. EMBO J. 1984 Jun;3(6):1429–1434. doi: 10.1002/j.1460-2075.1984.tb01988.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Talbot P., Almeida J. D. Human cytomegalovirus: purification of enveloped virions and dense bodies. J Gen Virol. 1977 Aug;36(2):345–349. doi: 10.1099/0022-1317-36-2-345. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. WATSON D. H., RUSSELL W. C., WILDY P. Electron microscopic particle counts on herpes virus using the phosphotungstate negative staining technique. Virology. 1963 Mar;19:250–260. doi: 10.1016/0042-6822(63)90062-x. [DOI] [PubMed] [Google Scholar]
  42. Weston K., Barrell B. G. Sequence of the short unique region, short repeats, and part of the long repeats of human cytomegalovirus. J Mol Biol. 1986 Nov 20;192(2):177–208. doi: 10.1016/0022-2836(86)90359-1. [DOI] [PubMed] [Google Scholar]

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