Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1989 Sep;63(9):3792–3800. doi: 10.1128/jvi.63.9.3792-3800.1989

Identification and characterization of a human cytomegalovirus gene coding for a membrane protein that is conserved among human herpesviruses.

R Lehner 1, H Meyer 1, M Mach 1
PMCID: PMC250972  PMID: 2547996

Abstract

A rabbit antiserum was raised against envelope material from purified human cytomegalovirus strain AD169. The serum recognized polypeptides 200, 170, 160, 75, 58, and 45 kilodaltons in size. It was used to screen a cDNA library constructed from poly(A)+ RNA from human cytomegalovirus-infected cells in the expression vector lambda gt11. A recombinant bacteriophage expressing cytomegalovirus-specific sequences was identified, and the corresponding gene was mapped to the HindIII R fragment. The gene is transcribed into a late 1.5-kilobase RNA. The nucleotide sequence of the coding region was determined. Computer analysis of the gene product revealed a polypeptide containing multiple potential membrane-spanning domains, representing a type of protein not identified in the envelope of herpesviruses before. The protein shows homology on the amino acid level to hypothetical proteins from reading frames BBRF3 of Epstein-Barr virus, UL10 of herpes simplex virus type 1, and ORF50 of varicella-zoster virus. By using an antiserum raised against procaryote-expressed parts of the cytomegalovirus membrane protein, a 45-kilodalton structural component of the virus was identified as the gene product.

Full text

PDF
3793

Images in this article

3794Image
on p.3794
3795Image
on p.3795
3795Image
on p.3795
3797Image
on p.3797
3797Image
on p.3797

Selected References

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

  1. Adams G. A., Rose J. K. Incorporation of a charged amino acid into the membrane-spanning domain blocks cell surface transport but not membrane anchoring of a viral glycoprotein. Mol Cell Biol. 1985 Jun;5(6):1442–1448. doi: 10.1128/mcb.5.6.1442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adams G. A., Rose J. K. Structural requirements of a membrane-spanning domain for protein anchoring and cell surface transport. Cell. 1985 Jul;41(3):1007–1015. doi: 10.1016/s0092-8674(85)80081-7. [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. Benko D. M., Gibson W. Primate cytomegalovirus glycoproteins: lectin-binding properties and sensitivities to glycosidases. J Virol. 1986 Sep;59(3):703–713. doi: 10.1128/jvi.59.3.703-713.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Birnstiel M. L., Busslinger M., Strub K. Transcription termination and 3' processing: the end is in site! Cell. 1985 Jun;41(2):349–359. doi: 10.1016/s0092-8674(85)80007-6. [DOI] [PubMed] [Google Scholar]
  6. Britt W. J., Auger D. Identification of a 65 000 dalton virion envelope protein of human cytomegalovirus. Virus Res. 1985 Dec;4(1):31–36. doi: 10.1016/0168-1702(85)90018-8. [DOI] [PubMed] [Google Scholar]
  7. Britt W. J. Neutralizing antibodies detect a disulfide-linked glycoprotein complex within the envelope of human cytomegalovirus. Virology. 1984 Jun;135(2):369–378. doi: 10.1016/0042-6822(84)90193-4. [DOI] [PubMed] [Google Scholar]
  8. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  9. Chee M., Rudolph S. A., Plachter B., Barrell B., Jahn G. Identification of the major capsid protein gene of human cytomegalovirus. J Virol. 1989 Mar;63(3):1345–1353. doi: 10.1128/jvi.63.3.1345-1353.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Claesson-Welsh L., Spear P. G. Amino-terminal sequence, synthesis, and membrane insertion of glycoprotein B of herpes simplex virus type 1. J Virol. 1987 Jan;61(1):1–7. doi: 10.1128/jvi.61.1.1-7.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Craighead J. E., Kanich R. E., Almeida J. D. Nonviral microbodies with viral antigenicity produced in cytomegalovirus-infected cells. J Virol. 1972 Oct;10(4):766–775. doi: 10.1128/jvi.10.4.766-775.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Davison A. J., Taylor P. Genetic relations between varicella-zoster virus and Epstein-Barr virus. J Gen Virol. 1987 Apr;68(Pt 4):1067–1079. doi: 10.1099/0022-1317-68-4-1067. [DOI] [PubMed] [Google Scholar]
  15. Debroy C., Pederson N., Person S. Nucleotide sequence of a herpes simplex virus type 1 gene that causes cell fusion. Virology. 1985 Aug;145(1):36–48. doi: 10.1016/0042-6822(85)90199-0. [DOI] [PubMed] [Google Scholar]
  16. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Farrar G. H., Oram J. D. Characterization of the human cytomegalovirus envelope glycoproteins. J Gen Virol. 1984 Nov;65(Pt 11):1991–2001. doi: 10.1099/0022-1317-65-11-1991. [DOI] [PubMed] [Google Scholar]
  19. Fleckenstein B., Müller I., Collins J. Cloning of the complete human cytomegalovirus genome in cosmids. Gene. 1982 Apr;18(1):39–46. doi: 10.1016/0378-1119(82)90054-3. [DOI] [PubMed] [Google Scholar]
  20. Gibson W. Protein counterparts of human and simian cytomegaloviruses. Virology. 1983 Jul 30;128(2):391–406. doi: 10.1016/0042-6822(83)90265-9. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Hu N., Messing J. The making of strand-specific M13 probes. Gene. 1982 Mar;17(3):271–277. doi: 10.1016/0378-1119(82)90143-3. [DOI] [PubMed] [Google Scholar]
  23. Irmiere A., Gibson W. Isolation of human cytomegalovirus intranuclear capsids, characterization of their protein constituents, and demonstration that the B-capsid assembly protein is also abundant in noninfectious enveloped particles. J Virol. 1985 Oct;56(1):277–283. doi: 10.1128/jvi.56.1.277-283.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Jahn G., Kouzarides T., Mach M., Scholl B. C., Plachter B., Traupe B., Preddie E., Satchwell S. C., Fleckenstein B., Barrell B. G. Map position and nucleotide sequence of the gene for the large structural phosphoprotein of human cytomegalovirus. J Virol. 1987 May;61(5):1358–1367. doi: 10.1128/jvi.61.5.1358-1367.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Jahn G., Scholl B. C., Traupe B., Fleckenstein B. The two major structural phosphoproteins (pp65 and pp150) of human cytomegalovirus and their antigenic properties. J Gen Virol. 1987 May;68(Pt 5):1327–1337. doi: 10.1099/0022-1317-68-5-1327. [DOI] [PubMed] [Google Scholar]
  26. Kanich R. E., Craighead J. E. Human cytomegalovirus infection of cultured fibroblasts. II. Viral replicative sequence of a wild and an adapted strain. Lab Invest. 1972 Sep;27(3):273–282. [PubMed] [Google Scholar]
  27. 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]
  28. Kouzarides T., Bankier A. T., Satchwell S. C., Preddy E., Barrell B. G. An immediate early gene of human cytomegalovirus encodes a potential membrane glycoprotein. Virology. 1988 Jul;165(1):151–164. doi: 10.1016/0042-6822(88)90668-x. [DOI] [PubMed] [Google Scholar]
  29. Kouzarides T., Bankier A. T., Satchwell S. C., Weston K., Tomlinson P., Barrell B. G. Large-scale rearrangement of homologous regions in the genomes of HCMV and EBV. Virology. 1987 Apr;157(2):397–413. doi: 10.1016/0042-6822(87)90282-0. [DOI] [PubMed] [Google Scholar]
  30. Kozak M. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res. 1984 Jan 25;12(2):857–872. doi: 10.1093/nar/12.2.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Landini M. P., Severi B., Badiali L., Gonczol E., Mirolo G. Structural components of human cytomegalovirus: in situ localization of the major glycoprotein. Intervirology. 1987;27(3):154–160. doi: 10.1159/000149734. [DOI] [PubMed] [Google Scholar]
  34. Landini M. P., Severi B., Furlini G., Badiali De Giorgi L. Human cytomegalovirus structural components: intracellular and intraviral localization of p28 and p65-69 by immunoelectron microscopy. Virus Res. 1987 Jul;8(1):15–23. doi: 10.1016/0168-1702(87)90036-0. [DOI] [PubMed] [Google Scholar]
  35. Liebowitz D., Wang D., Kieff E. Orientation and patching of the latent infection membrane protein encoded by Epstein-Barr virus. J Virol. 1986 Apr;58(1):233–237. doi: 10.1128/jvi.58.1.233-237.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. MCGAVRAN M. H., SMITH M. G. ULTRASTRUCTURAL, CYTOCHEMICAL, AND MICROCHEMICAL OBSERVATIONS ON CYTOMEGALOVIRUS (SALIVARY GLAND VIRUS) INFECTION OF HUMAN CELLS IN TISSUE CULTURE. Exp Mol Pathol. 1965 Feb;76:1–10. doi: 10.1016/0014-4800(65)90019-5. [DOI] [PubMed] [Google Scholar]
  37. Mach M., Utz U., Fleckenstein B. Mapping of the major glycoprotein gene of human cytomegalovirus. J Gen Virol. 1986 Jul;67(Pt 7):1461–1467. doi: 10.1099/0022-1317-67-7-1461. [DOI] [PubMed] [Google Scholar]
  38. 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]
  39. McGeoch D. J., Dolan A., Frame M. C. DNA sequence of the region in the genome of herpes simplex virus type 1 containing the exonuclease gene and neighbouring genes. Nucleic Acids Res. 1986 Apr 25;14(8):3435–3448. doi: 10.1093/nar/14.8.3435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. McGeoch D. J. On the predictive recognition of signal peptide sequences. Virus Res. 1985 Oct;3(3):271–286. doi: 10.1016/0168-1702(85)90051-6. [DOI] [PubMed] [Google Scholar]
  41. Meyer H., Bankier A. T., Landini M. P., Brown C. M., Barrell B. G., Rüger B., Mach M. Identification and procaryotic expression of the gene coding for the highly immunogenic 28-kilodalton structural phosphoprotein (pp28) of human cytomegalovirus. J Virol. 1988 Jul;62(7):2243–2250. doi: 10.1128/jvi.62.7.2243-2250.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Meyers J. D., Flournoy N., Thomas E. D. Nonbacterial pneumonia after allogeneic marrow transplantation: a review of ten years' experience. Rev Infect Dis. 1982 Nov-Dec;4(6):1119–1132. doi: 10.1093/clinids/4.6.1119. [DOI] [PubMed] [Google Scholar]
  43. Murray R. J., Wang D., Young L. S., Wang F., Rowe M., Kieff E., Rickinson A. B. Epstein-Barr virus-specific cytotoxic T-cell recognition of transfectants expressing the virus-coded latent membrane protein LMP. J Virol. 1988 Oct;62(10):3747–3755. doi: 10.1128/jvi.62.10.3747-3755.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Nowak B., Sullivan C., Sarnow P., Thomas R., Bricout F., Nicolas J. C., Fleckenstein B., Levine A. J. Characterization of monoclonal antibodies and polyclonal immune sera directed against human cytomegalovirus virion proteins. Virology. 1984 Jan 30;132(2):325–338. doi: 10.1016/0042-6822(84)90039-4. [DOI] [PubMed] [Google Scholar]
  45. 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]
  46. 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]
  47. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Schreier P. H., Cortese R. A fast and simple method for sequencing DNA cloned in the single-stranded bacteriophage M13. J Mol Biol. 1979 Mar 25;129(1):169–172. doi: 10.1016/0022-2836(79)90068-8. [DOI] [PubMed] [Google Scholar]
  49. Spector S. A., Hirata K. K., Newman T. R. Identification of multiple cytomegalovirus strains in homosexual men with acquired immunodeficiency syndrome. J Infect Dis. 1984 Dec;150(6):953–956. doi: 10.1093/infdis/150.6.953. [DOI] [PubMed] [Google Scholar]
  50. Stinski M. F. Human cytomegalovirus: glycoproteins associated with virions and dense bodies. J Virol. 1976 Aug;19(2):594–609. doi: 10.1128/jvi.19.2.594-609.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Stinski M. F. Synthesis of proteins and glycoproteins in cells infected with human cytomegalovirus. J Virol. 1977 Sep;23(3):751–767. doi: 10.1128/jvi.23.3.751-767.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. 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]
  53. Tanford C. The hydrophobic effect and the organization of living matter. Science. 1978 Jun 2;200(4345):1012–1018. doi: 10.1126/science.653353. [DOI] [PubMed] [Google Scholar]
  54. Thorley-Lawson D. A., Israelsohn E. S. Generation of specific cytotoxic T cells with a fragment of the Epstein-Barr virus-encoded p63/latent membrane protein. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5384–5388. doi: 10.1073/pnas.84.15.5384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. 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]
  56. Wilson I. A., Skehel J. J., Wiley D. C. Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3 A resolution. Nature. 1981 Jan 29;289(5796):366–373. doi: 10.1038/289366a0. [DOI] [PubMed] [Google Scholar]
  57. Xu-Bin, Murayama T., Ishida K., Furukawa T. Characterization of IgG Fc receptors induced by human cytomegalovirus. J Gen Virol. 1989 Apr;70(Pt 4):893–900. doi: 10.1099/0022-1317-70-4-893. [DOI] [PubMed] [Google Scholar]
  58. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  59. Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES