1887

Abstract

We have investigated the human cytomegalovirus (HCMV) US22 gene family members UL23, UL24, UL43 and US22. Specific antibodies were generated to identify pUL23 (33 kDa), pUL24 (40 kDa) and pUL43 (48 kDa), while pUS22 was identified by monoclonal antibody HWLF1. A C-terminally truncated UL43 product (pUL43t; 21 kDa) produced by a deletion mutant was also investigated. The UL24 and UL43 genes were expressed with early-late (γ1) and true-late (γ2) kinetics, respectively. Immunoblot and immuno-EM studies demonstrated that pUL23, pUL24, pUL43 and pUS22 were virion tegument components. Immunofluorescence and immuno-EM studies showed that pUL23, pUL24, pUL43 and pUL43t were located in cytoplasmic protein aggregates, manifesting two forms: complex juxtanuclear structures and smaller, membrane-bound aggregates resembling dense bodies. The complex-type aggregate is a putative site of particle maturation. Because pUL43t was present in protein aggregates, but under-represented in virus particles compared to pUL43, it was concluded that N-terminal sequences target pUL43 to protein aggregates and that C-terminal sequences are important for incorporation into particles. Since three other US22 family products (pUL36, pTRS1 and pIRS1) are documented tegument components, at least seven of the twelve US22 family genes encode tegument proteins, suggesting that the products of the remaining five genes might be similarly located. These findings demonstrate a common biological feature among most, if not all, US22 family proteins and implicate the family in events occurring immediately after virus penetration.

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2002-06-01
2019-10-15
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References

  1. Bahr, U. & Darai, G. ( 2001; ). Analysis and characterization of the complete genome of Tupaia (tree shrew) herpesvirus. Journal of Virology 75, 4854-4870.[CrossRef]
    [Google Scholar]
  2. Brown, J. M., Kaneshima, H. & Mocarski, E. S. ( 1995; ). Dramatic interstrain differences in the replication of human cytomegalovirus in SCID-hu mice. Journal of Infectious Diseases 171, 1599-1603.[CrossRef]
    [Google Scholar]
  3. Butcher, S. J., Aitken, J., Mitchell, J., Gowen, B. & Dargan, D. J. ( 1998; ). Structure of the human cytomegalovirus B capsid by electron cryomicroscopy and image reconstruction. Journal of Structural Biology 124, 70-76.[CrossRef]
    [Google Scholar]
  4. Chambers, J., Angulo, A., Amaratunga, D., Guo, Y. Y., Wan, J. S., Bittner, A., Frueh, K., Jackson, M. R., Peterson, P. A., Erlander, M. G. & Ghazal, P. ( 1999; ). DNA microarrays of the complex human cytomegalovirus genome: profiling kinetic class with drug sensitivity of viral gene expression. Journal of Virology 73, 5757-5766.
    [Google Scholar]
  5. Chee, M. S., Bankier, A. T., Beck, S., Bohni, R., Brown, C. M., Cerny, R., Horsnell, T., Hutchinson, C. A., Kouzarides, T., Martignetti, J. A., Preddie, E., Satchwell, S. C., Tomlinson, P., Weston, K. M. & Barrell, B. G. ( 1990; ). Analysis of the protein coding content of the sequence of the human cytomegalovirus strain AD169. Current Topics in Microbiology and Immunology 154, 125-170.
    [Google Scholar]
  6. Dalton, A. J. ( 1975; ). Microvesicles and vesicles of multivesiculate bodies versus ‘‘virus like’’ particles. Journal of the National Cancer Institute 54, 1137-1147.
    [Google Scholar]
  7. Dargan, D. J., Jamieson, F. E., Maclean, J., Dolan, A., Addison, C. & McGeoch, D. J. ( 1997; ). The published DNA sequence of the human cytomegalovirus strain AD169 lacks 929 base pairs affecting genes UL42 and UL43. Journal of Virology 71, 9833-9836.
    [Google Scholar]
  8. Efstathiou, S., Lawrence, G. L., Brown, C. M. & Barrell, B. C. ( 1992; ). Identification of homologues to the human cytomegalovirus US22 gene family in human herpesvirus 6. Journal of General Virology 73, 1661-1671.[CrossRef]
    [Google Scholar]
  9. Geng, Y., Chandran, B., Josephs, S. F. & Wood, C. ( 1992; ). Identification and characterization of a human herpesvirus 6 gene segment that transactivates the human immunodeficiency virus type 1 promoter. Journal of Virology 66, 1564-1570.
    [Google Scholar]
  10. Gompels, U. A., Nicholas, J., Lawrence, G., Jones, M., Thomson, B. J., Martin, M. E. D., Efstathiou, S., Craxton, M. & Macaulay, H. A. ( 1995; ). The DNA sequence of human herpesvirus-6: structure, coding content and genome evolution. Virology 209, 29-51.[CrossRef]
    [Google Scholar]
  11. Hensel, G., Meyer, H., Gartner, S., Brand, G. & Kern, H. F. ( 1995; ). Nuclear localization of the human cytomegalovirus tegument protein pp150 (ppUL32). Journal of General Virology 76, 1591-1601.[CrossRef]
    [Google Scholar]
  12. Irmiere, A. & Gibson, W. ( 1983; ). Isolation and characterization of a noninfectious virion-like particle released from cells infected with the human strains of cytomegalovirus. Virology 130, 118-133.[CrossRef]
    [Google Scholar]
  13. Jones, T. R. & Muzithras, V. P. ( 1992; ). A cluster of dispensable genes within the human cytomegalovirus genome short component: IRS1, US1, through US5 and the US6 family. Journal of Virology 66, 2541-2546.
    [Google Scholar]
  14. Kashanchi, F., Thompson, J., Sadaie, M. R., Doniger, J., Duvall, J., Brady, J. N. & Rosenthal, L. J. ( 1994; ). Transcriptional activation of minimal HIV-1 promoter by ORF-1 protein expressed from the Sal1-L fragment of human herpesvirus 6. Virology 201, 95-106.[CrossRef]
    [Google Scholar]
  15. Kerry, J. A., Priddy, M. A., Jervey, T. Y., Kohler, C. P., Staley, T. L., Vanson, C. D., Jones, T. R., Iskenderian, A. C., Anders, D. A. & Stenberg, R. M. ( 1996; ). Multiple regulatory events influence human cytomegalovirus DNA polymerase (UL54) expression during viral infection. Journal of Virology 70, 373-382.
    [Google Scholar]
  16. Kouzarides, T., Bankier, A. T., Satchwell, S. C., Preddy, E. & Barrell, B. G. ( 1988; ). An immediate early gene of human cytomegalovirus encodes a potential membrane glycoprotein. Virology 165, 151-164.[CrossRef]
    [Google Scholar]
  17. Landini, M. P., Severi, B., Furlini, G. & De Giorgi, L. B. ( 1987; ). Human cytomegalovirus structural components: intracellular and intraviral localization of p28 and p65–69 by immunoelectron microscopy. Virus Research 8, 15-23.[CrossRef]
    [Google Scholar]
  18. Megaw, A. G., Rapaport, D., Avidor, B., Frenkel, N. & Davison, A. J. ( 1998; ). The DNA sequence of the RK strain of human herpesvirus 7. Virology 244, 119-132.[CrossRef]
    [Google Scholar]
  19. Mocarski, E. S., Pereira, L. & McCormick, L. A. ( 1988; ). Human cytomegalovirus ICP22, the product of the HWLF1 reading frame, is an early nuclear protein that is released from cells. Journal of General Virology 69, 2613-2621.[CrossRef]
    [Google Scholar]
  20. Mori, Y., Yagi, H., Shimamoto, T., Sunagawa, T., Inagi, R., Kondo, K., Tano, Y. & Yamanishi, K. ( 1998; ). Analysis of human herpesvirus 6 U3 gene, which is a positional homolog of human cytomegalovirus UL24. Virology 249, 129-139.[CrossRef]
    [Google Scholar]
  21. Nicholas, J. ( 1996; ). Determination and analysis of the complete nucleotide sequence of human herpesvirus 7. Journal of Virology 70, 5975-5989.
    [Google Scholar]
  22. Nicholas, J. & Martin, M. ( 1994; ). Nucleotide sequence of a 38·5-kilobase-pair region of the genome of human herpesvirus 6 encoding human cytomegalovirus immediate-early gene homologues and transactivating functions. Journal of Virology 68, 597-610.
    [Google Scholar]
  23. Oram, J. D., Downing, R. G., Akrigg, A., Dollery, A. A., Duggleby, C. J., Wilkinson, G. W. & Greenaway, P. J. ( 1982; ). Use of recombinant plasmids to investigate the structure of the human cytomegalovirus genome. Journal of General Virology 59, 111-129.[CrossRef]
    [Google Scholar]
  24. Patterson, C. E. & Shenk, T. ( 1999; ). Human cytomegalovirus UL36 protein is dispensable for viral replication in cultured cells. Journal of Virology 73, 7126-7131.
    [Google Scholar]
  25. Rawlinson, W. D., Farrell, H. E. & Barrell, B. G. ( 1996; ). Analysis of the complete DNA sequence of murine cytomegalovirus. Journal of Virology 70, 8833-8849.
    [Google Scholar]
  26. Romanowski, M. J. & Shenk, T. ( 1997; ). Characterization of the human cytomegalovirus irs1 and trs1 genes: a second immediate-early transcription unit within irs1 whose product antagonizes transcriptional activation. Journal of Virology 71, 1485-1496.
    [Google Scholar]
  27. Romanowski, M. J., Garrido-Guerrero, E. & Shenk, T. ( 1997; ). pIRS1 and pTRS1 are present in human cytomegalovirus virions. Journal of Virology 72, 5703-5705.
    [Google Scholar]
  28. Sanchez, V., Greis, K. D., Sztul, E. & Britt, W. J. ( 2000; ). Accumulation of virion tegument and envelope proteins in a stable cytoplasmic compartment during human cytomegalovirus replication: characterization of a potential site of virus assembly. Journal of Virology 74, 975-986.[CrossRef]
    [Google Scholar]
  29. Severi, B., Landini, M. P., Cenacchi, G., Zini, N. & Maraldi, N. M. ( 1992; ). Human cytomegalovirus nuclear and cytoplasmic dense bodies. Archives of Virology 123, 193-207.[CrossRef]
    [Google Scholar]
  30. Skaletskaya, A., Bartle, L. M., Chittenden, T., McCormick, A. L., Mocarski, E. S. & Goldmacher, V. S. ( 2001; ). A cytomegalovirus-encoded inhibitor of apoptosis that suppresses caspase-8 activation. Proceedings of the National Academy of Sciences, USA 98, 7829-7834.[CrossRef]
    [Google Scholar]
  31. Stannard, L. & Hardie, D. R. ( 1991; ). An Fc receptor for human immunoglobulin G is located within the tegument of human cytomegalovirus. Journal of General Virology 65, 3411-3415.
    [Google Scholar]
  32. Stasiak, P. C. & Mocarski, E. S. ( 1992; ). Transactivation of the cytomegalovirus ICP36 gene promoter requires the α gene product TRS1 in addition to IE1 and IE2. Journal of Virology 66, 1050-1058.
    [Google Scholar]
  33. Tenney, D. J. & Colberg, P. A. ( 1991; ). Expression of the human cytomegalovirus UL36–38 immediate early gene region during permissive infection. Virology 182, 199-210.[CrossRef]
    [Google Scholar]
  34. Vink, C., Beuken, E. & Bruggeman, C. A. ( 2000; ). Complete DNA sequence of the rat cytomegalovirus genome. Journal of Virology 74, 7656-7665.[CrossRef]
    [Google Scholar]
  35. Xiao, J., Tong, T., Zhan, X., Haghjoo, E. & Liu, F. ( 2000; ). In vitro and in vivo characterization of a murine cytomegalovirus with a transposon insertional mutation at open reading frame M43. Journal of Virology 74, 9488-9497.[CrossRef]
    [Google Scholar]
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