1887

Abstract

Human cytomegalovirus phosphoprotein pp65 is targeted to the cell nucleus immediately after infection. Deletion and point mutation analysis of the pp65 gene expressed in insect cells showed that two hydrophilic regions (HP1 and HP2) within the pp65 C-terminal 40% each harboured an independent nuclear localization signal (NLS); strong association to the nuclear stroma also requires the N-terminal domain. Either region, when fused to chloramphenicol acetyltransferase, localized the reporter protein to the nucleus in insect cells as well as in NIH 3T3 cells and human lung fibroblasts. In addition, HP1 was found to be the target of pp65 Ser/Thr phosphorylation in insect cells and a prokaryotically expressed HP1 was actively phosphorylated by casein kinase II, for which two site clusters map in HP1. These findings indicate that pp65 includes two NLSs, one of which has the potential to be modulated by phosphorylation.

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1996-06-01
2024-12-08
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References

  1. Britt W. J., Auger D. 1986; Human cytomegalovirus virion-associated protein with kinase activity. Journal of Virology 59:185–188
    [Google Scholar]
  2. Britt W. J., Vugler L. 1987; Structural and immunological characterization of the intracellular forms of an abundant 68000 M r human cytomegalovirus protein. Journal of General Virology 68:1897–1907
    [Google Scholar]
  3. Dingwall C., Laskey R. A. 1991; Nuclear targeting sequences – a consensus?. Trends in Biochemical Sciences 16:478–481
    [Google Scholar]
  4. Grefte A., Harmsen M. C., van der Giessen M., Knollema S., van Son W. J., The T. H. 1994; Presence of human cytomegalovirus (HCMV) immediate early mRNA but not ppUL83 (lower matrix protein pp65) mRNA in polymorphonuclear and mononuclear leukocytes during active HCMV infection. Journal of General Virology 75:1989–1998
    [Google Scholar]
  5. Grefte J. M. M., van der Gun B. T. F., Schmolke S., van der Giessen M., van Son W. J., Plachter B., Jahn G., The T. H. 1992; The lower matrix protein pp65 is the principal viral antigen present in peripheral blood leukocytes during an active cytomegalovirus infection. Journal of General Virology 73:2923–2932
    [Google Scholar]
  6. Jahn G., Kouzarides T., Mach M., Scholl B.-C., Plachter B., Traupe B., Preddie E., Satchwell S. C., Fleckenstein B., Barrell B. G. 1987; Map position and nucleotide sequence of the gene for the large structural phosphoprotein of human cytomegalovirus. Journal of Virology 61:1358–1367
    [Google Scholar]
  7. Landini M. P., Severi B., Furlini G., Badiali L. 1987; HCMV structural components: intracellular and intraviral localization of p28 and p65-69 by immunoelectron microscopy. Virus Research 8:15–23
    [Google Scholar]
  8. Michelson S., Tardy-Panit M., Barzu O. 1985; Catalytic properties of a human cytomegalovirus-induced protein kinase. European Journal of Biochemistry 149:393–399
    [Google Scholar]
  9. Miyamoto C., Smith G. E., Farrell-Towt J., Chizzonite R., Summers M. D., Ju G. 1985; Production of human c-myc protein in insect cells infected with a baculovirus expression vector. Molecular and Cellular Biology 5:2860–2865
    [Google Scholar]
  10. Pande H., Lee T. D., Churchill M. A., Zaia J. A. 1990; Structural analysis of a 64-kDa major structural protein of human cytomegalovirus (Towne): identification of a phosphorylation site and comparison to pp65 of HCMV (ADI 69). Virology 178:6–14
    [Google Scholar]
  11. Pinna L. A. 1990; Casein kinase 2: an ‘eminence grise’ in cellular regulation?. Biochimica et Biophysica Acta 1054:267–284
    [Google Scholar]
  12. Revello M. G., Percivalle E., Di Matteo A., Morini F., Gerna G. 1992; Nuclear expression of the lower matrix protein of human cytomegalovirus in peripheral blood leukocytes of immunocompromised viraemic patients. Journal of General Virology 73:437–442
    [Google Scholar]
  13. Roby C., Gibson W. 1986; Characterization of phosphoproteins and protein kinase activity of virions, noninfectious enveloped particles, and dense bodies of human cytomegalovirus. Journal of Virology 59:714–727
    [Google Scholar]
  14. Schmolke S., Kern H. F., Drescher P., Jahn G., Plachter B. 1995a; The dominant phosphoprotein pp65 (UL83) of human cytomegalovirus is dispensable for growth in cell culture. Journal of Virology 69:5959–5968
    [Google Scholar]
  15. Schmolke S., Drescher P., Jahn G., Plachter B. 1995a; Nuclear targeting of the tegument protein pp65 (UL83) of human cytomegalovirus: an unusual bipartite nuclear localization signal functions with other portions of the protein to mediate its efficient nuclear transport. Journal of Virology 69:1071–1078
    [Google Scholar]
  16. Somogy T., Michelson S., Masse M. O. 1990; Genomic location of a human cytomegalovirus protein with protein kinase activity (PK68). Virology 174:276–285
    [Google Scholar]
  17. Spaete R. R., Gehrz R. C., Landini M. P. 1994; Human cytomegalovirus structural proteins. Journal of General Virology 75:3287–3308
    [Google Scholar]
  18. Wang Y. 1992; Fluorescence microscopic analysis of cytoskeletal organization and dynamics. In The Cytoskeleton – A Practical Approach pp. 1–22 Edited by Carraway L. K., Carraway A. C. New York: IRL Press;
    [Google Scholar]
  19. Weiner D., Gibson W. 1981; Identification of a primate CMV group-common protein antigen. Virology 115:182–191
    [Google Scholar]
  20. Weiner D., Gibson W., Fields K. L. 1985; Anti-complement immunofluorescence establishes nuclear localization of human cytomegalovirus matrix protein. Virology 147:19–28
    [Google Scholar]
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