1887

Abstract

The characterization is reported of the human herpesvirus-6B (HHV-6B) rep/U94 gene, which is a homologue of the adeno-associated virus type 2 rep. In this study, a monoclonal antibody was produced against HHV-6B REP (anti-REP mAb). Immunofluorescence staining using the anti-REP mAb showed that REP was localized to the nucleus in HHV-6-infected MT4 cells. It was first detected at 24 h post-infection (p.i.) and accumulated to higher levels by 72 h p.i. REP may be expressed only at very low levels in HHV-6-infected cells: even when the late protein glycoprotein H was detected in nearly 90% of HHV-6-infected cells, REP was detected in only a small percentage of them. Western blot analysis showed that the anti-REP mAb recognized a 56-kDa polypeptide in HHV-6B-infected MT4 cells. Furthermore, the REP protein was shown to bind single-stranded DNA.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-83-4-847
2002-04-01
2020-10-27
Loading full text...

Full text loading...

/deliver/fulltext/jgv/83/4/0830847a.html?itemId=/content/journal/jgv/10.1099/0022-1317-83-4-847&mimeType=html&fmt=ahah

References

  1. Ablashi D.. 1993; Human herpesvirus-6 strain groups: a nomenclature. Archives of Virology129:363–366
    [Google Scholar]
  2. Antoni B. A., Rabson A. B., Miller I. L., Trempe J. P., Chejanovsky N., Carter B. J.. 1991; Adeno-associated virus Rep protein inhibits human immunodeficiency virus type 1 production in human cells. Journal of Virology65:396–404
    [Google Scholar]
  3. Beaton A., Palumbo P., Berns K. I.. 1989; Expression from the adeno-associated virus p5 and p19 promoters is negatively regulated in trans by the rep protein. Journal of Virology63:4450–4454
    [Google Scholar]
  4. Chiorini J. A., Weitzman M. D., Owens R. A., Urcelay E., Safer B., Kotin R. M.. 1994; Biologically active Rep proteins of adeno-associated virus type 2 produced as fusion proteins in Escherichia coli . Journal of Virology68:797–804
    [Google Scholar]
  5. Daibata M., Taguchi T., Kamioka M., Kubonishi I., Taguchi H., Miyoshi I.. 1998; Identification of integrated human herpesvirus 6 DNA in early pre-B cell acute lymphoblastic leukemia. Leukemia12:1002–1004
    [Google Scholar]
  6. Dominguez G., Dambaugh T. R., Stamey F. R., Dewhurst S., Inoue N., Pellett P. E.. 1999; Human herpesvirus 6B genome sequence: coding content and comparison with human herpesvirus 6A. Journal of Virology73:8040–8052
    [Google Scholar]
  7. Ge H., Roeder R. G.. 1994; Purification, cloning, and characterization of a human coactivator, PC4, that mediates transcriptional activation of class II genes. Cell78:513–523
    [Google Scholar]
  8. Gompels U. A., Nicholas J., Lawrence G., Jones M., Thomson B. J., Martin M. E., Efstathiou S., Craxton M., Macaulay H. A.. 1995; The DNA sequence of human herpesvirus-6: structure, coding content, and genome evolution. Virology209:29–51
    [Google Scholar]
  9. Heilbronn R., Burkle A., Stephan S., zur Hausen H.. 1990; The adeno-associated virus rep gene suppresses herpes simplex virus-induced DNA amplification. Journal of Virology64:3012–3018
    [Google Scholar]
  10. Hermonat P. L.. 1994a; Adeno-associated virus inhibits human papillomavirus type 16: a viral interaction implicated in cervical cancer. Cancer Research54:2278–2281
    [Google Scholar]
  11. Hermonat P. L.. 1994b; Down-regulation of the human c-fos and c-myc proto-oncogene promoters by adeno-associated virus Rep78. Cancer Letters81:129–136
    [Google Scholar]
  12. Hermonat P. L., Santin A. D., Batchu R. B.. 1996; The adeno-associated virus Rep78 major regulatory/transformation suppressor protein binds cellular Sp1 in vitro and evidence of a biological effect. Cancer Research56:5299–5304
    [Google Scholar]
  13. Hermonat P. L., Santin A. D., Carter C. A., Parham G. P., Quirk J. G.. 1997; Multiple cellular proteins are recognized by the adeno-associated virus Rep78 major regulatory protein and the amino-half of Rep78 is required for many of these interactions. Biochemistry and Molecular Biology International43:409–420
    [Google Scholar]
  14. Hermonat P. L., Santin A. D., Batchu R. B., Zhan D.. 1998; The adeno-associated virus Rep78 major regulatory protein binds the cellular TATA-binding protein in vitro and in vivo . Virology245:120–127
    [Google Scholar]
  15. Im D. S., Muzyczka N.. 1990; The AAV origin binding protein Rep68 is an ATP-dependent site-specific endonuclease with DNA helicase activity. Cell61:447–457
    [Google Scholar]
  16. Im D. S., Muzyczka N.. 1992; Partial purification of adeno-associated virus Rep78, Rep52, and Rep40 and their biochemical characterization. Journal of Virology66:1119–1128
    [Google Scholar]
  17. Isegawa Y., Mukai T., Nakano K., Kagawa M., Chen J., Mori Y., Sunagawa T., Kawanishi K., Sashihara J., Hata A., Zou P., Kosuge H., Yamanishi K.. 1999; Comparison of the complete DNA sequences of human herpesvirus 6 variants A and B. Journal of Virology73:8053–8063
    [Google Scholar]
  18. Kaiser K., Stelzer G., Meisterernst M.. 1995; The coactivator p15 (PC4) initiates transcriptional activation during TFIIA-TFIID-promoter complex formation. EMBO Journal14:3520–3527
    [Google Scholar]
  19. Kellermann O. K., Ferenci T.. 1982; Maltose-binding protein from Escherichia coli . Methods in Enzymology90:459–463
    [Google Scholar]
  20. Kyostio S. R., Wonderling R. S., Owens R. A.. 1995; Negative regulation of the adeno-associated virus (AAV) P5 promoter involves both the P5 rep binding site and the consensus ATP-binding motif of the AAV Rep68 protein. Journal of Virology69:6787–6796
    [Google Scholar]
  21. Labow M. A., Hermonat P. L., Berns K. I.. 1986; Positive and negative autoregulation of the adeno-associated virus type 2 genome. Journal of Virology60:251–258
    [Google Scholar]
  22. Labow M. A., Graf L. H. Jr, Berns K. I.. 1987; Adeno-associated virus gene expression inhibits cellular transformation by heterologous genes. Molecular and Cellular Biology7:1320–1325
    [Google Scholar]
  23. Lee T. I., Young R. A.. 1998; Regulation of gene expression by TBP-associated proteins. Genes & Development12:1398–1408
    [Google Scholar]
  24. Linden R. M., Ward P., Giraud C., Winocour E., Berns K. I.. 1996; Site-specific integration by adeno-associated virus. Proceedings of the National Academy of Sciences, USA93:11288–11294
    [Google Scholar]
  25. Luppi M., Marasca R., Barozzi P., Ferrari S., Ceccherini-Nelli L., Batoni G., Merelli E., Torelli G.. 1993; Three cases of human herpesvirus-6 latent infection: integration of viral genome in peripheral blood mononuclear cell DNA. Journal of Medical Virology40:44–52
    [Google Scholar]
  26. Luppi M., Barozzi P., Marasca R., Torelli G.. 1994; Integration of human herpesvirus-6 (HHV-6) genome in chromosome 17 in two lymphoma patients. Leukemia8:S41–S45
    [Google Scholar]
  27. Mori Y., Yagi H., Shimamoto T., Isegawa Y., 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 UL 24 gene. Virology249:129–139
    [Google Scholar]
  28. Mori Y., Dhepakson P., Shimamoto T., Ueda K., Gomi Y., Tani H., Matsuura Y., Yamanishi K.. 2000; Expression of human herpesvirus 6B rep within infected cells and binding of its gene product to the TATA-binding protein in vitro and in vivo . Journal of Virology74:6096–6104
    [Google Scholar]
  29. Ni T. H., Zhou X., McCarty D. M., Zolotukhin I., Muzyczka N.. 1994; In vitro replication of adeno-associated virus DNA. Journal of Virology68:1128–1138
    [Google Scholar]
  30. Okuno T., Sao H., Asada H., Shiraki K., Takahashi M., Yamanishi K.. 1990; Analysis of a glycoprotein of human herpesvirus 6 (HHV-6) using monoclonal antibodies. Virology176:625–628
    [Google Scholar]
  31. Rapp J. C., Krug L. T., Inoue N., Dambaugh T. R., Pellett P. E.. 2000; U94, the human herpesvirus 6 homolog of the parvovirus nonstructural gene, is highly conserved among isolates and is expressed at low mRNA levels as a spliced transcript. Virology268:504–516
    [Google Scholar]
  32. Rotola A., Ravaioli T., Gonelli A., Dewhurst S., Cassai E., Di Luca D.. 1998; U94 of human herpesvirus 6 is expressed in latently infected peripheral blood mononuclear cells and blocks viral gene expression in transformed lymphocytes in culture. Proceedings of the National Academy of Sciences, USA95:13911–13916
    [Google Scholar]
  33. Salahuddin S. Z., Ablashi D. V., Markham P. D., Josephs S. F., Sturzenegger S., Kaplan M., Halligan G., Biberfeld P., Wong-Staal F., Kramarsky B.. and others 1986; Isolation of a new virus, HBLV, in patients with lymphoproliferative disorders. Science234:596–601
    [Google Scholar]
  34. Tanaka H., Yoshimura Y., Nozaki M., Yomogida K., Tsuchida J., Tosaka Y., Habu T., Nakanishi T., Okada M., Nojima H., Nishimune Y.. 1999; Identification and characterization of a haploid germ cell-specific nuclear protein kinase (Haspin) in spermatid nuclei and its effects on somatic cells. Journal of Biological Chemistry274:17049–17057
    [Google Scholar]
  35. Thomson B. J., Efstathiou S., Honess R. W.. 1991; Acquisition of the human adeno-associated virus type-2 rep gene by human herpesvirus type-6. Nature351:78–80
    [Google Scholar]
  36. Thomson B. J., Weindler F. W., Gray D., Schwaab V., Heilbronn R.. 1994; Human herpesvirus 6 (HHV-6) is a helper virus for adeno-associated virus type 2 (AAV-2) and the AAV-2 rep gene homologue in HHV-6 can mediate AAV-2 DNA replication and regulate gene expression. Virology204:304–311
    [Google Scholar]
  37. Torelli G., Barozzi P., Marasca R., Cocconcelli P., Merelli E., Ceccherini-Nelli L., Ferrari S., Luppi M.. 1995; Targeted integration of human herpesvirus 6 in the p arm of chromosome 17 of human peripheral blood mononuclear cells in vivo. Journal of Medical Virology46:178–188
    [Google Scholar]
  38. Vink C., Beuken E., Bruggeman C. A.. 2000; Complete DNA sequence of the rat cytomegalovirus genome. Journal of Virology74:7656–7665
    [Google Scholar]
  39. Weger S., Wendland M., Kleinschmidt J. A., Heilbronn R.. 1999; The adeno-associated virus type 2 regulatory proteins rep78 and rep68 interact with the transcriptional coactivator PC4. Journal of Virology73:260–269
    [Google Scholar]
  40. Yamanishi K., Okuno T., Shiraki K., Takahashi M., Kondo T., Asano Y., Kurata T.. 1988; Identification of human herpesvirus-6 as a causal agent for exanthem subitum. Lancet i:1065–1067
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-83-4-847
Loading
/content/journal/jgv/10.1099/0022-1317-83-4-847
Loading

Data & Media loading...

Most cited this month 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