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. 1996 Sep;70(9):6097–6105. doi: 10.1128/jvi.70.9.6097-6105.1996

Proteins associated with purified human cytomegalovirus particles.

C J Baldick Jr 1, T Shenk 1
PMCID: PMC190631  PMID: 8709233

Abstract

Virion-associated proteins isolated from purified human cytomegalovirus particles (strain AD169) were used as substrates for chemical sequence analysis. Extracellular virions, noninfectious enveloped particles, and dense bodies were purified by negative viscosity-positive density gradient centrifugation, and their component proteins were separated by denaturing polyacrylamide gel electrophoresis. The deduced amino acid sequence of individual protein bands was used to identify six corresponding viral genes whose products have not previously been identified as virion constituents: UL47, UL25, UL88, UL85, UL26, and UL48.5. In addition, a 45-kDa cellular protein was identified, and the protein fragments sequenced have a high degree of amino acid identity with actin. However, antiactin monoclonal and polyclonal antibodies did not react with a specific protein in the virus preparations, suggesting that this 45-kDa protein is an immunologically distinct isoform of actin. The newly identified viral and cellular proteins were resistant to protease treatment of purified virions, suggesting that they are unlikely to be contaminants of the viral preparations.

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Selected References

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  1. Bradshaw P. A., Duran-Guarino M. R., Perkins S., Rowe J. I., Fernandez J., Fry K. E., Reyes G. R., Young L., Foung S. K. Localization of antigenic sites on human cytomegalovirus virion structural proteins encoded by UL48 and UL56. Virology. 1994 Nov 15;205(1):321–328. doi: 10.1006/viro.1994.1648. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. 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]
  4. Dallas P. B., Lyons P. A., Hudson J. B., Scalzo A. A., Shellam G. R. Identification and characterization of a murine cytomegalovirus gene with homology to the UL25 open reading frame of human cytomegalovirus. Virology. 1994 May 1;200(2):643–650. doi: 10.1006/viro.1994.1227. [DOI] [PubMed] [Google Scholar]
  5. Damsky C. H., Sheffield J. B., Tuszynski G. P., Warren L. Is there a role for actin in virus budding? J Cell Biol. 1977 Nov;75(2 Pt 1):593–605. doi: 10.1083/jcb.75.2.593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Davis M. G., Huang E. S. Nucleotide sequence of a human cytomegalovirus DNA fragment encoding a 67-kilodalton phosphorylated viral protein. J Virol. 1985 Oct;56(1):7–11. doi: 10.1128/jvi.56.1.7-11.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Davison M. D., Rixon F. J., Davison A. J. Identification of genes encoding two capsid proteins (VP24 and VP26) of herpes simplex virus type 1. J Gen Virol. 1992 Oct;73(Pt 10):2709–2713. doi: 10.1099/0022-1317-73-10-2709. [DOI] [PubMed] [Google Scholar]
  8. Desai P., DeLuca N. A., Glorioso J. C., Person S. Mutations in herpes simplex virus type 1 genes encoding VP5 and VP23 abrogate capsid formation and cleavage of replicated DNA. J Virol. 1993 Mar;67(3):1357–1364. doi: 10.1128/jvi.67.3.1357-1364.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fiala M., Honess R. W., Heiner D. C., Heine J. W., Jr, Murnane J., Wallace R., Guze L. B. Cytomegalovirus proteins. I. Polypeptides of virions and dense bodies. J Virol. 1976 Jul;19(1):243–254. doi: 10.1128/jvi.19.1.243-254.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Geballe A. P., Leach F. S., Mocarski E. S. Regulation of cytomegalovirus late gene expression: gamma genes are controlled by posttranscriptional events. J Virol. 1986 Mar;57(3):864–874. doi: 10.1128/jvi.57.3.864-874.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gibson W., Clopper K. S., Britt W. J., Baxter M. K. Human cytomegalovirus (HCMV) smallest capsid protein identified as product of short open reading frame located between HCMV UL48 and UL49. J Virol. 1996 Aug;70(8):5680–5683. doi: 10.1128/jvi.70.8.5680-5683.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gibson W., Marcy A. I., Comolli J. C., Lee J. Identification of precursor to cytomegalovirus capsid assembly protein and evidence that processing results in loss of its carboxy-terminal end. J Virol. 1990 Mar;64(3):1241–1249. doi: 10.1128/jvi.64.3.1241-1249.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Gibson W. Structural and nonstructural proteins of strain Colburn cytomegalovirus. Virology. 1981 Jun;111(2):516–537. doi: 10.1016/0042-6822(81)90354-8. [DOI] [PubMed] [Google Scholar]
  15. Giuffre R. M., Tovell D. R., Kay C. M., Tyrrell D. L. Evidence for an interaction between the membrane protein of a paramyxovirus and actin. J Virol. 1982 Jun;42(3):963–968. doi: 10.1128/jvi.42.3.963-968.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Goins W. F., Stinski M. F. Expression of a human cytomegalovirus late gene is posttranscriptionally regulated by a 3'-end-processing event occurring exclusively late after infection. Mol Cell Biol. 1986 Dec;6(12):4202–4213. doi: 10.1128/mcb.6.12.4202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Grefte J. M., van der Gun B. T., Schmolke S., van der Giessen M., van Son W. J., Plachter B., Jahn G., The T. H. The lower matrix protein pp65 is the principal viral antigen present in peripheral blood leukocytes during an active cytomegalovirus infection. J Gen Virol. 1992 Nov;73(Pt 11):2923–2932. doi: 10.1099/0022-1317-73-11-2923. [DOI] [PubMed] [Google Scholar]
  18. Grundy J. E., McKeating J. A., Griffiths P. D. Cytomegalovirus strain AD169 binds beta 2 microglobulin in vitro after release from cells. J Gen Virol. 1987 Mar;68(Pt 3):777–784. doi: 10.1099/0022-1317-68-3-777. [DOI] [PubMed] [Google Scholar]
  19. Grundy J. E., McKeating J. A., Ward P. J., Sanderson A. R., Griffiths P. D. Beta 2 microglobulin enhances the infectivity of cytomegalovirus and when bound to the virus enables class I HLA molecules to be used as a virus receptor. J Gen Virol. 1987 Mar;68(Pt 3):793–803. doi: 10.1099/0022-1317-68-3-793. [DOI] [PubMed] [Google Scholar]
  20. Herman I. M. Actin isoforms. Curr Opin Cell Biol. 1993 Feb;5(1):48–55. doi: 10.1016/s0955-0674(05)80007-9. [DOI] [PubMed] [Google Scholar]
  21. Irmiere A., Gibson W. Isolation and characterization of a noninfectious virion-like particle released from cells infected with human strains of cytomegalovirus. Virology. 1983 Oct 15;130(1):118–133. doi: 10.1016/0042-6822(83)90122-8. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. Jahn G., Mach M. Human cytomegalovirus phosphoproteins and glycoproteins and their coding regions. Curr Top Microbiol Immunol. 1990;154:171–185. doi: 10.1007/978-3-642-74980-3_7. [DOI] [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. Kim K. S., Sapienza V. J., Carp R. I., Moon H. M. Analysis of structural polypeptides of purified human cytomegalovirus. J Virol. 1976 Dec;20(3):604–611. doi: 10.1128/jvi.20.3.604-611.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Klages S., Rüger B., Jahn G. Multiplicity dependent expression of the predominant phosphoprotein pp65 of human cytomegalovirus. Virus Res. 1989 Feb;12(2):159–168. doi: 10.1016/0168-1702(89)90061-0. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Lahijani R. S., Otteson E. W., Adlish J. D., St Jeor S. C. Characterization of a human cytomegalovirus 1.6-kilobase late mRNA and identification of its putative protein product. J Virol. 1991 Jan;65(1):373–381. doi: 10.1128/jvi.65.1.373-381.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lahijani R. S., Otteson E. W., St Jeor S. C. A possible role for nonsense suppression in the synthesis of a human cytomegalovirus 58-kDa virion protein. Virology. 1992 Jan;186(1):309–312. doi: 10.1016/0042-6822(92)90087-6. [DOI] [PubMed] [Google Scholar]
  31. Landini M. P., Michelson S. Human cytomegalovirus proteins. Prog Med Virol. 1988;35:152–185. [PubMed] [Google Scholar]
  32. 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]
  33. Lees-Miller J. P., Helfman D. M., Schroer T. A. A vertebrate actin-related protein is a component of a multisubunit complex involved in microtubule-based vesicle motility. Nature. 1992 Sep 17;359(6392):244–246. doi: 10.1038/359244a0. [DOI] [PubMed] [Google Scholar]
  34. Liu B., Stinski M. F. Human cytomegalovirus contains a tegument protein that enhances transcription from promoters with upstream ATF and AP-1 cis-acting elements. J Virol. 1992 Jul;66(7):4434–4444. doi: 10.1128/jvi.66.7.4434-4444.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. McKeating J. A., Griffiths P. D., Grundy J. E. Cytomegalovirus in urine specimens has host beta 2 microglobulin bound to the viral envelope: a mechanism of evading the host immune response? J Gen Virol. 1987 Mar;68(Pt 3):785–792. doi: 10.1099/0022-1317-68-3-785. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Muhua L., Karpova T. S., Cooper J. A. A yeast actin-related protein homologous to that in vertebrate dynactin complex is important for spindle orientation and nuclear migration. Cell. 1994 Aug 26;78(4):669–679. doi: 10.1016/0092-8674(94)90531-2. [DOI] [PubMed] [Google Scholar]
  38. Naito S., Matsumoto S. Identification of cellular actin within the rabies virus. Virology. 1978 Nov;91(1):151–163. doi: 10.1016/0042-6822(78)90363-x. [DOI] [PubMed] [Google Scholar]
  39. Orvell C. Structural polypeptides of mumps virus. J Gen Virol. 1978 Dec;41(3):527–539. doi: 10.1099/0022-1317-41-3-527. [DOI] [PubMed] [Google Scholar]
  40. Plamann M., Minke P. F., Tinsley J. H., Bruno K. S. Cytoplasmic dynein and actin-related protein Arp1 are required for normal nuclear distribution in filamentous fungi. J Cell Biol. 1994 Oct;127(1):139–149. doi: 10.1083/jcb.127.1.139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Rixon F. J., Davison M. D., Davison A. J. Identification of the genes encoding two capsid proteins of herpes simplex virus type 1 by direct amino acid sequencing. J Gen Virol. 1990 May;71(Pt 5):1211–1214. doi: 10.1099/0022-1317-71-5-1211. [DOI] [PubMed] [Google Scholar]
  42. Robb M. J., Wilson M. A., Vierula P. J. A fungal actin-related protein involved in nuclear migration. Mol Gen Genet. 1995 Jun 10;247(5):583–590. doi: 10.1007/BF00290350. [DOI] [PubMed] [Google Scholar]
  43. Robson L., Gibson W. Primate cytomegalovirus assembly protein: genome location and nucleotide sequence. J Virol. 1989 Feb;63(2):669–676. doi: 10.1128/jvi.63.2.669-676.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Roby C., Gibson W. Characterization of phosphoproteins and protein kinase activity of virions, noninfectious enveloped particles, and dense bodies of human cytomegalovirus. J Virol. 1986 Sep;59(3):714–727. doi: 10.1128/jvi.59.3.714-727.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Rüger B., Klages S., Walla B., Albrecht J., Fleckenstein B., Tomlinson P., Barrell B. Primary structure and transcription of the genes coding for the two virion phosphoproteins pp65 and pp71 of human cytomegalovirus. J Virol. 1987 Feb;61(2):446–453. doi: 10.1128/jvi.61.2.446-453.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Sagara J., Tsukita S., Yonemura S., Tsukita S., Kawai A. Cellular actin-binding ezrin-radixin-moesin (ERM) family proteins are incorporated into the rabies virion and closely associated with viral envelope proteins in the cell. Virology. 1995 Jan 10;206(1):485–494. doi: 10.1016/s0042-6822(95)80064-6. [DOI] [PubMed] [Google Scholar]
  47. Sarov I., Abady I. The morphogenesis of human cytomegalovirus. Isolation and polypeptide characterization of cytomegalovirions and dense bodies. Virology. 1975 Aug;66(2):464–473. doi: 10.1016/0042-6822(75)90218-4. [DOI] [PubMed] [Google Scholar]
  48. Schmolke S., Kern H. F., Drescher P., Jahn G., Plachter B. The dominant phosphoprotein pp65 (UL83) of human cytomegalovirus is dispensable for growth in cell culture. J Virol. 1995 Oct;69(10):5959–5968. doi: 10.1128/jvi.69.10.5959-5968.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Schroer T. A., Fyrberg E., Cooper J. A., Waterston R. H., Helfman D., Pollard T. D., Meyer D. I. Actin-related protein nomenclature and classification. J Cell Biol. 1994 Dec;127(6 Pt 2):1777–1778. doi: 10.1083/jcb.127.6.1777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Somogyi T., Michelson S., Masse M. J. Genomic location of a human cytomegalovirus protein with protein kinase activity (PK68). Virology. 1990 Jan;174(1):276–285. doi: 10.1016/0042-6822(90)90075-3. [DOI] [PubMed] [Google Scholar]
  51. Spaete R. R., Gehrz R. C., Landini M. P. Human cytomegalovirus structural proteins. J Gen Virol. 1994 Dec;75(Pt 12):3287–3308. doi: 10.1099/0022-1317-75-12-3287. [DOI] [PubMed] [Google Scholar]
  52. Spear G. T., Lurain N. S., Parker C. J., Ghassemi M., Payne G. H., Saifuddin M. Host cell-derived complement control proteins CD55 and CD59 are incorporated into the virions of two unrelated enveloped viruses. Human T cell leukemia/lymphoma virus type I (HTLV-I) and human cytomegalovirus (HCMV). J Immunol. 1995 Nov 1;155(9):4376–4381. [PubMed] [Google Scholar]
  53. Stannard L. M. Beta 2 microglobulin binds to the tegument of cytomegalovirus: an immunogold study. J Gen Virol. 1989 Aug;70(Pt 8):2179–2184. doi: 10.1099/0022-1317-70-8-2179. [DOI] [PubMed] [Google Scholar]
  54. Stannard L. M., Hardie D. R. An Fc receptor for human immunoglobulin G is located within the tegument of human cytomegalovirus. J Virol. 1991 Jun;65(6):3411–3415. doi: 10.1128/jvi.65.6.3411-3415.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. 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]
  56. Tatman J. D., Preston V. G., Nicholson P., Elliott R. M., Rixon F. J. Assembly of herpes simplex virus type 1 capsids using a panel of recombinant baculoviruses. J Gen Virol. 1994 May;75(Pt 5):1101–1113. doi: 10.1099/0022-1317-75-5-1101. [DOI] [PubMed] [Google Scholar]
  57. Tyrrell D. L., Norrby E. Structural polypeptides of measles virus. J Gen Virol. 1978 May;39(2):219–229. doi: 10.1099/0022-1317-39-2-219. [DOI] [PubMed] [Google Scholar]
  58. Vandekerckhove J., Weber K. At least six different actins are expressed in a higher mammal: an analysis based on the amino acid sequence of the amino-terminal tryptic peptide. J Mol Biol. 1978 Dec 25;126(4):783–802. doi: 10.1016/0022-2836(78)90020-7. [DOI] [PubMed] [Google Scholar]
  59. Welch A. R., McNally L. M., Gibson W. Cytomegalovirus assembly protein nested gene family: four 3'-coterminal transcripts encode four in-frame, overlapping proteins. J Virol. 1991 Aug;65(8):4091–4100. doi: 10.1128/jvi.65.8.4091-4100.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Wright J. F., Kurosky A., Pryzdial E. L., Wasi S. Host cellular annexin II is associated with cytomegalovirus particles isolated from cultured human fibroblasts. J Virol. 1995 Aug;69(8):4784–4791. doi: 10.1128/jvi.69.8.4784-4791.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Wykes M. N., Price P., Shellam G. R. The effects of beta-2-microglobulin on the infectivity of murine cytomegalovirus. Arch Virol. 1992;123(1-2):59–72. doi: 10.1007/BF01317138. [DOI] [PubMed] [Google Scholar]
  62. Yamauchi M., Nishiyama Y., Fujioka H., Isomura S., Maeno K. On the intracellular transport and the nuclear association of human cytomegalovirus structural proteins. J Gen Virol. 1985 Apr;66(Pt 4):675–684. doi: 10.1099/0022-1317-66-4-675. [DOI] [PubMed] [Google Scholar]
  63. Yao F., Courtney R. J. Association of ICP0 but not ICP27 with purified virions of herpes simplex virus type 1. J Virol. 1992 May;66(5):2709–2716. doi: 10.1128/jvi.66.5.2709-2716.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

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