1887

Journal of General Virology header logo

Volume 75, Issue 12

Intracellular localization and DNA-binding activity of a class of viral early phosphoproteins in human fibroblasts infected with human cytomegalovirus (Towne strain) Free

Abstract

Indirect immunofluorescence (IF) with monoclonal antibody M23 prepared against the nuclei of human embryo lung (HEL) cells infected with human cytomegalovirus (HCMV) Towne strain showed that the M23 antigen reactive with the M23 antibody was localized within distinct foci throughout the nucleus of infected HEL cells shortly after infection, even at 2 h post-infection (p.i.). The foci increased in size by 24 h p.i. and then the IF patterns changed to show the nuclear inclusion body-like structures at 72 h p.i. Treatment with phosphono-acetic acid, a HCMV DNA replication inhibitor, resulted in a nuclear pattern similar to that observed shortly after infection. The double-labelled IF test revealed that the HCMV UL44 antigen essential for viral DNA replication colocalized with the M23 antigen in the same intranuclear structure shortly after infection whereas neither viral antigen appeared to colocalize in most cells later after infection. The M23 antibody immunoprecipitated four proteins, 34K, 43K, 50K and 84K, in infected cells. To examine whether these proteins correspond to four early phosphoproteins encoded by the HCMV strain AD169 genome, the Towne strain DNA sequence corresponding to that encoding both the 34K and 43K proteins of strain AD169 was determined and transiently expressed in COS-7, Vero and HEL cells. These proteins were detected by the M23 antibody within the foci of these nuclei as found in the nuclei of productively infected cells shortly after infection. In addition, the 34K, 43K and 50K proteins at least were shown to be DNA-binding proteins by double- and single-stranded DNA-cellulose column chromatography. The relationship of these proteins to the status of viral DNA replication is discussed.

  • Received:
  • Accepted:
  • Published Online:
© The Journal of General Virology 1994
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-75-12-3309
1994-12-01
2024-04-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/75/12/JV0750123309.html?itemId=/content/journal/jgv/10.1099/0022-1317-75-12-3309&mimeType=html&fmt=ahah

References

  1. Anders D. G., Kidd J. R., Gibson W. 1987; Immunological characterization of an early cytomegalovirus single-strand DNA binding protein with similarities to the HSV major DNA-binding protein. Virology 161:579–588
     [Google Scholar]
  2. Challberg M. D. 1986; A method for identifying the viral genes required for herpes DNA replication. Proceedings of the National Academy of Sciences, U.S.A 83:9094–9098
     [Google Scholar]
  3. Freemer C. S., Bertoni G., Takagi S., Sairenji T. 1993; A novel antigen associated with Epstein-Barr virus productive cycle. Virology 194:387–392
     [Google Scholar]
  4. Goodrich L. D., Schaffer P. A., Dorsky D. I., Crumpacker C. S., Parris D. S. 1990; Localization of the herpes simplex virus type 1 65-kilodalton DNA-binding protein and DNA polymerase in the presence and absence of viral DNA synthesis. Journal of Virology 64:5738–5749
     [Google Scholar]
  5. Graham F. L., Van Der Eb A. J. 1973; A new technique for the assay of infectivity of human adenovirus DNA. Virology 52:456–467
     [Google Scholar]
  6. Hirai K., Watanabe Y. 1976; Induction of a-type DNA polymerase in human cytomegalovirus-infected WI-38 cells. Bio-chimica et biophysica acta 447:328–339
     [Google Scholar]
  7. Hirai K., Furukawa T., Plotkin S. A. 1976; Induction of DNA polymerase in WI-38 and guinea pig cells infected with human cytomegalovirus (HCMV). Virology 70:251–255
     [Google Scholar]
  8. Hirai K., Maeda F., Watanabe Y. 1977; Expression of early virus functions in human cytomegalovirus infected HEL cells: effect of ultraviolet light-irradiation of the virus. Journal of General Virology 38:121–133
     [Google Scholar]
  9. Hirai K., Nakajima K., Ikuta K., Kirisawa R., Kawakami Y., Mikami T., Kato S. 1986; Similarities and dissimilarities in the structure and expression of viral genomes of various virus strains immunologically related to Marek’s disease virus. Archives of Virology 89:113–130
     [Google Scholar]
  10. Huang E. -S. 1975; Human cytomegalovirus. IV. Specific inhibition of virus-induced DNA polymerase activity and viral DNA replication by phosphonoacetic acid. Journal of Virology 16:1560–1565
     [Google Scholar]
  11. Ikuta K., Nishi Y., Kato S., Hirai K. 1981; Immunoprecipi-tation of Marek’s disease virus-specific polypeptides with chicken antibodies purified by affinity chromatography. Virology 114:277–281
     [Google Scholar]
  12. Ikuta K., Honma H., Maotani K., Ueda S., Kato S., Hirai K. 1982; Monoclonal antibodies specific to and cross-reactive with Marek’s disease virus and herpesvirus of turkeys. Bikens Journal 25:171–175
     [Google Scholar]
  13. Ikuta K., Ueda S., Kato S., Hirai K. 1983; Monoclonal antibodies reactive with the surface and secreted glycoproteins of Marek’s disease virus and herpesvirus of turkeys. Journal of General Virology 64:2597–2610
     [Google Scholar]
  14. Kawamura A., Aoyama Y. 1982 Immunofluorescence in Medical Science Tokyo: University of Tokyo Press;
     [Google Scholar]
  15. Kemble G. W., McCormick A. L., Pereira L., Mocarski E. S. 1987; A cytomegalovirus protein with properties of herpes simplex virus 1CP8: partial purification of the polypeptide and map position of the gene. Journal of Virology 61:3143–3151
     [Google Scholar]
  16. Kiehl A., Dorsky D. I. 1991; Cooperation of EBV DNA polymerase and EA-D (BMRF1) in vitro and colocalization in nuclei of infected cells. Virology 184:330–340
     [Google Scholar]
  17. Knipe D. M., Quinian M. P., Spang A. E. 1982; Characterization of two conformational forms of the major DNA binding protein encoded by herpes simplex virus. Journal of Virology 44:736–741
     [Google Scholar]
  18. Kops A. D., Knipe D. M. 1988; Formation of DNA replication structures in herpes virus-infected cells requires a viral DNA binding protein. Cell 55:857–868
     [Google Scholar]
  19. Lawrence G. L., Chee M., Craxton M. A., Gompels U. A., Honess R. W., Barrell B. G. 1990; Human herpesvirus 6 is closely related to human cytomegalovirus. Journal of Virology 64:287–299
     [Google Scholar]
  20. Marsden H. S., Campbell M. E. M., Haarr L., Frame M. C., Parris D. S., Murphy M., Hope R. G., Muller M. T., Preston C. M. 1987; The 65K DNA-binding and virion trans- inducing proteins of herpes simplex virus type 1. Journal of Virology 61:2428–2437
     [Google Scholar]
  21. Nakajima K., Ikuta K., Ueda S., Kato S., Hirai K. 1986; Identification with monoclonal antibodies of virus-specific DNA- binding proteins in the nuclei of cells infected with three serotypes of Marek’s disease virus-related viruses. Journal of Virology 59:154–158
     [Google Scholar]
  22. Pari G. S., Anders D. G. 1993; Eleven loci encoding trans-acting factors are required for transient complementation of human cytomegalovirus oriLyt-dependent DNA replication. Journal of Virology 67:6979–6988
     [Google Scholar]
  23. Pari G. S., Kacia M. A., Anders D. G. 1993; Open reading frames UL44, IRS1/TRS1, and UL36-38 are required for transient complementation of human cytomegalovirus oriLyt-dependent DNA synthesis. Journal of Virology 67:2575–2582
     [Google Scholar]
  24. Plachter B., Nordin M., Zweygberg W. B., Mach M., Stein H., Grillner L., Jahn G. 1992; The DNA-binding protein P52 of human cytomegalovirus reacts with monoclonal antibody CCH2 and associates with the nuclear membrane at late times after infection. Virus Research 24:265–276
     [Google Scholar]
  25. Quinlan M. P., Chen L. B., Knipe D. M. 1984; The intranuclear location of a herpes simplex virus DNA-binding protein is determined by the status of viral DNA replication. Cell 36:857–868
     [Google Scholar]
  26. Roizman B., Desrosiers R. C., Fleckenstein B., Lopez C., Minson A. C., Studdert M. J. 1992; The family Herpesviridae: an update. Archives of Virology 123:425–449
     [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, U.S.A 74:5463–5467
     [Google Scholar]
  28. Staprans S. I., Spector D. H. 1986; 2·2-Kilobase class of early transcripts encoded by cell-related sequences in human cytomegalovirus strain AD169. Journal of Virology 56:591–602
     [Google Scholar]
  29. Staprans S. I., Rabert D. K., Spector D. H. 1988; Identification of sequence requirements and trans-acting functions necessary for regulated expression of a human cytomegalovirus early gene. Journal of Virology 62:3463–3473
     [Google Scholar]
  30. Takagi S., Takada K., Sairenji T. 1991; Formation of intranuclear replication compartments of Epstein-Barr virus with redistribution of BZLFl and BMRF1 gene products. Virology 185:309–315
     [Google Scholar]
  31. Weir H. M., Calder J. M., Stow N. D. 1989; Binding of the herpes simplex virus type 1 UL19 gene product to an origin of viral DNA replication. Nucleic Acids Research 17:1409–1425
     [Google Scholar]
  32. Winkler M., Rice S. A., Stamminger T. 1994; UL69 of human cytomegalovirus, an open reading frame with homology to ICP27 of herpes simplex virus, encodes a transactivator of gene expression. Journal of Virology 68:3943–3954
     [Google Scholar]
  33. Wright D. A., Spector D. H. 1989; Post-transcriptional regulation of a class human cytomegalovirus phosphoproteins encoded by an early transcription unit. Journal of Virology 63:3117–3127
     [Google Scholar]
  34. Wright D. A., Staprans S. I., Spector D. H. 1988; Four phosphoproteins with common amino termini are encoded by human cytomegalovirus AD169. Journal of Virology 62:331–340
     [Google Scholar]
  35. Wu C. A., Nelson N. J., McGeoch D. J., Challberg M. D. 1988; Identification of herpes simplex virus type 1 genes required for origin-dependent synthesis. Journal of Virology 62:435–443
     [Google Scholar]
  36. Yamaguchi Y., Hironaka T., Kajiwara M., Tateno E., Kita H., Hirai K. 1992; Increased sensitivity for detection of human cytomegalovirus in urine by removal of inhibitors for the polymerase chain reaction. Journal of Virological Methods 37:209–218
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-75-12-3309
Loading
Intracellular localization and DNA-binding activity of a class of viral early phosphoproteins in human fibroblasts infected with human cytomegalovirus (Towne strain)
J. Gen. Virol. 75, 3309 (1994); https://doi.org/10.1099/0022-1317-75-12-3309
/content/journal/jgv/10.1099/0022-1317-75-12-3309
/content/journal/jgv/10.1099/0022-1317-75-12-3309
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error