1887

Abstract

The complete ORF54 of the Kaposi’s sarcoma-associated herpesvirus (KSHV) (human herpesvirus-8; HHV-8) was cloned and expressed in E. coli. The results show that KSHV/HHV-8 ORF54 encodes a functional dUTPase which specifically hydrolyses dUTP to dUMP. Monoclonal antibodies against the HHV-8 dUTPase detected a protein with the expected molecular mass of 35 kDa in HHV-8-infected BCBL-1 cells. Induction of the lytic replication cycle of HHV-8 by treatment of BCBL-1 cells with the phorbol ester TPA resulted in an increased expression of the protein which was not inhibited by phosphonoacetic acid, indicating that the protein is expressed early in the lytic replication cycle. Moreover, the sporadic expression of the HHV-8 dUTPase in tissue sections of Kaposi’s sarcoma was detected by immunohistochemistry.

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1999-05-01
2024-04-25
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References

  1. Björnberg O., Nyman P. O. 1996; The dUTPases from herpes simplex virus type 1 and mouse mammary tumour virus are less specific than the Escherichia coli enzyme. Journal of General Virology 77:3107–3111
    [Google Scholar]
  2. Boos H., Berger R., Kuklik Roos C., Iftner T., Mueller Lantzsch N. 1987; Enhancement of Epstein–Barr virus membrane protein (LMP) expression by serum, TPA, or n-butyrate in latently infected Raji cells. Virology 159:161–165
    [Google Scholar]
  3. Caradonna S. J., Adamkiewicz D. M. 1984; Purification and properties of the deoxyuridine triphosphate nucleotidohydrolase enzyme derived from HeLa S3 cells. Comparison to a distinct dUTP nucleotide-hydrolase induced in herpes simplex virus-infected HeLa S3 cells. Journal of Biological Chemistry 259:5459–5464
    [Google Scholar]
  4. Cattoretti G., Becker M. H., Key G., Duchrow M., Schluter C., Galle J., Gerdes J. 1992; Monoclonal antibodies against recombinant parts of the Ki-67 antigen (MIB 1 and MIB 3) detect proliferating cells in microwave–processed formalin-fixed paraffin sections. Journal of Pathology 168:357–363
    [Google Scholar]
  5. Cesarman E., Chang Y., Moore P. S., Said J. W., Knowles D. M. 1995; Kaposi′s sarcoma-associated herpesvirus-like DNA sequences in AIDS-related body-cavity-based lymphomas. New England Journal of Medicine 332:1186–1191
    [Google Scholar]
  6. Chang Y., Cesarman E., Pessin M. S., Lee F., Culpepper J., Knowles D. M., Moore P. S. 1994; Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi′s sarcoma. Science 266:1865–1869
    [Google Scholar]
  7. Climie S., Lutz T., Radul J., Sumner Smith M., Vandenberg E., McIntosh E. 1994; Expression of trimeric human dUTP pyro-phosphatase in Escherichia coli and purification of the enzyme. Protein Expression and Purification 5:252–258
    [Google Scholar]
  8. Cordell J. L., Falini B., Erber W. N., Ghosh A. K., Abdulaziz Z., MacDonald S., Pulford K. A., Stein H., Mason D. Y. 1984; Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes). Journal of Histochemistry and Cytochemistry 32:219–229
    [Google Scholar]
  9. el Hajj H. H., Zhang H., Weiss B. 1988; Lethality of a dut (deoxyuridine triphosphatase) mutation in Escherichia coli . Journal of Bacteriology 170:1069–1075
    [Google Scholar]
  10. Gadsden M. H., McIntosh E. M., Game J. C., Wilson P. J., Haynes R. H. 1993; dUTP pyrophosphatase is an essential enzyme in Saccharomyces cerevisiae . EMBO Journal 12:4425–4431
    [Google Scholar]
  11. Ganem D. 1997; KSHV and Kaposi′s sarcoma: the end of the beginning. Cell 91:157–160
    [Google Scholar]
  12. Gerdes J., Schwab U., Lemke H., Stein H. 1983; Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation. International Journal of Cancer 31:13–20
    [Google Scholar]
  13. Jöns A., Gerdts V., Lange E., Kaden V., Mettenleiter T. C. 1997; Attenuation of dUTPase-deficient pseudorabies virus for the natural host. Veterinary Microbiology 56:47–54
    [Google Scholar]
  14. Koerner T. J., Hill J. E., Myers A. M., Tzagoloff A. 1991; High-expression vectors with multiple cloning sites for construction of trpE fusion genes: pATH vectors. Methods in Enzymology 194:477–490
    [Google Scholar]
  15. Kremmer E., Kranz B., Hille A., Klein K., Eulitz M., Hoffmann-Fezer G., Feiden W., Herrmann K., Delecluse H.-J., Delsol G., Bornkamm G. W., Mueller-Lantzsch N., Grasser F. A. 1995; Rat monoclonal antibodies differentiating between the Epstein–Barr virus nuclear antigens 2A (EBNA2A) and 2B (EBNA2B). Virology 208:336–342
    [Google Scholar]
  16. Kremmer E., Sommer P., Grässer F. A. 1997; Epstein–Barr virus–encoded deoxyuridine triphosphate nucleotidohydrolase (dUTPase): a potential target for drug therapy. Transplantation Proceedings 29:812–814
    [Google Scholar]
  17. Larsson G., Nyman P. O., Kvassman J. O. 1996a; Kinetic characterization of dUTPase from Escherichia coli . Journal of Biological Chemistry 271:24010–24016
    [Google Scholar]
  18. Larsson G., Svensson L. A., Nyman P. O. 1996b; Crystal structure of the Escherichia coli dUTPase in complex with a substrate analogue (dUDP). Nature Structural Biology 3:532–538
    [Google Scholar]
  19. Lenoir G. M., Vuillaume M., Bonnardel C. 1985; The use of lymphomatous and lymphoblastoid cell lines in the study of Burkitt′s lymphoma. IARC Scientific Publications 60:309–318
    [Google Scholar]
  20. Lerner D. L., Wagaman P. C., Phillips T. R., Prospero Garcia O., Henriksen S. J., Fox H. S., Bloom F. E., Elder J. H. 1995; Increased mutation frequency of feline immunodeficiency virus lacking functional deoxyuridine triphosphatase. Proceedings of the National Academy of Sciences, USA 92:7480–7484
    [Google Scholar]
  21. Lichtenstein D. L., Rushlow K. E., Cook R. F., Raabe M. L., Swardson C. J., Kociba G. J., Issel C. J., Montelaro R. C. 1995; Replication in vitro and in vivo of an equine infectious anemia virus mutant deficient in dUTPase activity. Journal of Virology 69:2881–2888
    [Google Scholar]
  22. McGeoch D. J. 1990; Protein sequence comparisons show that the ′pseudoproteases′ encodedby poxviruses and certain retroviruses belong to the deoxyuridine triphosphatase family. Nucleic Acids Research 18:4105–4110
    [Google Scholar]
  23. Miller G., Lipman M. 1973; Release of infectious Epstein–Barr virus by transformed marmoset leukocytes. Proceedings of the National Academy of Sciences, USA 70:190–194
    [Google Scholar]
  24. Mol C. D., Harris J. M., McIntosh E. M., Tainer J. A. 1996; Human dUTP pyrophosphatase : uracil recognition by a beta hairpin and active sites formed by three separate subunits. Structure 4:1077–1092
    [Google Scholar]
  25. Neipel F., Albrecht J. C., Fleckenstein B. 1997; Cell-homologous genes in the Kaposi′s sarcoma-associated rhadinovirus human herpesvirus 8: determinants of its pathogenicity ?. Journal of Virology 71:4187–4192
    [Google Scholar]
  26. Nicholls J. M., Sommer P., Kremmer E., Ong K. S., Fung K., Lee J. M. F., Ng M. N., Grasser F. A. 1998; A new lytic antibody, 7D6, detects Epstein–Barr virus dUTPase in nonkeratinizing undifferentiated nasopharyngeal carcinoma. Laboratory Investigation 78:1031–1032
    [Google Scholar]
  27. Niedobitek G., Young L. S., Lau R., Brooks L., Greenspan D., Greenspan J. S., Rickinson A. B. 1991; Epstein–Barr virus infection in oral hairy leukoplakia: virus replication in the absence of a detectable latent phase. Journal of General Virology 72:3035–3046
    [Google Scholar]
  28. Niedobitek G., Kremmer E., Herbst H., Whitehead L., Dawson C. W., Niedobitek E., von Ostau C., Rooney N., Grasser F. A., Young L. S. 1997; Immunohistochemical detection of the Epstein–Barr virus– encoded latent membrane protein 2A in Hodgkin′s disease and infectious mononucleosis. Blood 90:1664–1672
    [Google Scholar]
  29. Nord J., Larsson G., Kvassman J. O., Rosengren A. M., Nyman P. O. 1997; dUTPase from the retrovirus equine infectious anemia virus: specificity, turnover and inhibition. FEBS Letters 414:271–274
    [Google Scholar]
  30. Pyles R. B., Sawtell N. M., Thompson R. L. 1992; Herpes simplex virus type 1 dUTPase mutants are attenuated for neurovirulence, neuroinvasiveness, and reactivation from latency. Journal of Virology 66:6706–6713
    [Google Scholar]
  31. Renne R., Lagunoff M., Zhong W., Ganem D. 1996a; The size and conformation of Kaposi′s sarcoma-associated herpesvirus (human herpesvirus 8) DNA in infected cells and virions. Journal of Virology 70:8151–8154
    [Google Scholar]
  32. Renne R., Zhong W., Herndier B., McGrath M., Abbey N., Kedes D., Ganem D. 1996b; Lytic growth of Kaposi′s sarcoma-associated herpesvirus (human herpesvirus 8) in culture. Nature Medicine 2:342–346
    [Google Scholar]
  33. Ross J., Williams M., Cohen J. I. 1997; Disruption of the varicella-zoster virus dUTPase and the adjacent ORF9A gene results in impaired growth and reduced syncytia formation in vitro . Virology 234:186–195
    [Google Scholar]
  34. Sarid R., Flore O., Bohenzky R. A., Chang Y., Moore P. S. 1998; Transcription mapping of the Kaposi′s sarcoma-associated herpesvirus (human herpesvirus 8) genome in a body cavity-based lymphoma cell line (BC-1). Journal of Virology 72:1005–1012
    [Google Scholar]
  35. Shlomai J., Kornberg A. 1978; Deoxyuridine triphosphatase of Escherichia coli. Purification, properties, and use as a reagent to reduce uracil incorporation into DNA. Journal of Biological Chemistry 253:3305–3312
    [Google Scholar]
  36. Sommer P., Kremmer E., Bier S., König S., Zalud P., Zeppezauer M., Jones J. F., Mueller-Lantzsch N., Grasser F. A. 1996; Cloning and expression of the Epstein–Barr virus-encoded dUTPase : patients with acute, reactivated or chronic virus infection develop antibodies against the enzyme. Journal of General Virology 77:2795–2805
    [Google Scholar]
  37. Soulier J., Grollet L., Oksenhendler E., Cacoub P., Cazals Hatem D., Babinet P., d’Agay M. F., Clauvel J. P., Raphael M., Degos L., Sigaux F. 1995; Kaposi′s sarcoma-associated herpesvirus-like DNA sequences in multicentric Castleman’s disease. Blood 86:1276–1280
    [Google Scholar]
  38. Staskus K. A., Zhong W., Gebhard K., Herndier B., Wang H., Renne R., Beneke J., Pudney J., Anderson D. J., Ganem D., Haase A. T. 1997; Kaposi′s sarcoma-associated herpesvirus gene expression in endothelial (spindle) tumor cells. Journal of Virology 71:715–719
    [Google Scholar]
  39. Steagall W. K., Robek M. D., Perry S. T., Fuller F. J., Payne S. L. 1995; Incorporation of uracil into viral DNA correlates with reduced replication of EIAV in macrophages. Virology 210:302–313
    [Google Scholar]
  40. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. 1990; Use of T7 RNA polymerase to direct expression of cloned genes. Methods in Enzymology 185:60–89
    [Google Scholar]
  41. Stürzl M., Blasig C., Schreier A., Neipel F., Hohenadl C., Cornali E., Ascherl G., Esser S., Brockmeyer N. H., Ekman M., Kaaya E. E., Tschachler E., Biberfeld P. 1997; Expression of HHV-8 latency-associated T0.7 RNA in spindle cells and endothelial cells of AIDS-associated, classical and African Kaposi′s sarcoma. International Journal of Cancer 72:68–71
    [Google Scholar]
  42. Williams M. V. 1988; Herpes simplex virus–induced dUTPase: target site for antiviral chemotherapy. Virology 166:262–264
    [Google Scholar]
  43. Zur Hausen H., O’Neill F. J., Freese U. K., Hecker E. 1978; Persisting oncogenic herpesvirus induced by the tumour promotor TPA. Nature 272:373–375
    [Google Scholar]
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