1887

Abstract

We have previously observed that the expression of two thymidylate biosynthesis enzymes, dihydrofolate reductase and thymidylate synthase (TS), is upregulated in quiescent human fibroblasts infected with human cytomegalovirus (HCMV). Here, we have demonstrated that HCMV increases expression of the cellular deoxycytidylate deaminase (dCMP deaminase), which provides the substrate for TS by converting dCMP to dUMP. We observed an increase in dCMP deaminase protein levels, whereas deoxyuridine triphosphatase (dUTPase), another cellular enzyme that may provide dUMP by hydrolysing dUTP, was undetectable. The essential requirement of cellular dCMP deaminase for productive HCMV replication was further emphasized by showing that a precursor of a potent dCMP deaminase inhibitor, zebularine, suppressed virus replication and DNA synthesis. These results suggest that HCMV exploits the host's dCMP deaminase activity to replicate in quiescent cells.

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2003-06-01
2020-01-18
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References

  1. Aduma, P. J., Gupta, S. V. & De Clercq, E. ( 1990; ). Antiherpes virus activity and effect on deoxyribonucleotide triphosphate pools of (E)-5-(2-bromovinyl)-2′-deoxycytidine in combination with deaminase inhibitors. Antivir Res 13, 111–126.
    [Google Scholar]
  2. Aduma, P. J., Gupta, S. V., Stuart, A. L. & Tourigny, G. ( 1991; ). Deoxyribonucleotide triphosphate pools of herpes simplex virus infected cells: the influence of selective antiherpes agents and the role of deaminase pathway. Biochem Cell Biol 69, 409–414.[CrossRef]
    [Google Scholar]
  3. Barchi, J. J., Jr, Cooney, D. A., Hao, Z., Weinberg, M., Taft, C., Marquez, V. E. & Ford, H., Jr ( 1995; ). Improved synthesis of zebularine [1-(β-d-ribofuranosyl)-dihydropyrimidin-2-one) nucleotides as inhibitors of human deoxycytidylate deaminase. J Enzym Inhib 9, 147–162.[CrossRef]
    [Google Scholar]
  4. Bissinger, A. L., Sinzger, C., Kaiserling, E. & Jahn, G. ( 2002; ). Human cytomegalovirus as a direct pathogen: correlation of multiorgan involvement and cell distribution with clinical and pathological findings in a case of congenital inclusion disease. J Med Virol 67, 200–206.[CrossRef]
    [Google Scholar]
  5. Chee, M. S., Bankier, A. T., Beck, S. & 12 other authors ( 1990; ). Analysis of the protein-coding content of human cytomegalovirus strain AD169. Curr Top Microbiol Immunol 154, 125–169.
    [Google Scholar]
  6. Fortunato, E. A., McElroy, A. K., Sanchez, V. & Spector, D. H. ( 2000; ). Exploitation of cellular signaling and regulatory pathways by human cytomegalovirus. Trends Microbiol 8, 111–119.[CrossRef]
    [Google Scholar]
  7. Gribaudo, G., Riera, L., Lembo, D., De Andrea, M., Gariglio, M., Rudge, T. L., Johnson, L. F. & Landolfo, S. ( 2000; ). Murine cytomegalovirus stimulates cellular thymidylate synthase gene expression in quiescent cells and requires the enzyme for replication. J Virol 74, 4979–4987.[CrossRef]
    [Google Scholar]
  8. Gribaudo, G., Riera, L., Rudge, T. L., Caposio, P., Johnson, L. F. & Landolfo, S. ( 2002; ). Human cytomegalovirus infection induces cellular thymidylate synthase gene expression in quiescent fibroblasts. J Gen Virol 83, 2983–2993.
    [Google Scholar]
  9. Hatanaka, M. & Dulbecco, R. ( 1966; ). Induction of DNA synthesis by SV40. Proc Natl Acad Sci U S A 56, 736–740.[CrossRef]
    [Google Scholar]
  10. Hengel, H., Brune, W. & Koszinowski, U. H. ( 1998; ). Immune evasion by cytomegalovirus: survival strategies of a highly adapted opportunist. Trends Microbiol 6, 190–197.[CrossRef]
    [Google Scholar]
  11. Johnson, L. F. ( 1994; ). Posttranscriptional regulation of thymidylate synthase gene expression. J Cell Biochem 54, 387–392.[CrossRef]
    [Google Scholar]
  12. Kuhn, K., Bertling, W. M. & Emmrich, F. ( 1993; ). Cloning of a functional cDNA for human cytidine deaminse (CDD) and its use as a marker of monocyte/macrophage differentiation. Biochem Biophys Res Commun 190, 1–7.[CrossRef]
    [Google Scholar]
  13. Ladner, R. D., McNulty, D. E., Carr, S. A., Roberts, G. D. & Caradonna, S. J. ( 1996; ). Characterization of distinct nuclear and mitochondrial forms of human deoxyuridine triphosphate nucleotidohydrolase. J Biol Chem 271, 7745–7751.[CrossRef]
    [Google Scholar]
  14. Landolfo, S., Gariglio, M., Gribaudo, G. & Lembo, D. ( 2002; ). The human cytomegalovirus. Pharmacol Ther (in press).
    [Google Scholar]
  15. Lembo, D., Angeretti, A., Gariglio, M. & Landolfo, S. ( 1998; ). Murine cytomegalovirus induces expression and enzyme activity of dihydrofolate reductase in quiescent cells. J Gen Virol 78, 2803–2808.
    [Google Scholar]
  16. Lembo, D., Gribaudo, G., Cavallo, R., Riera, L., Angeretti, A., Hertel, L. & Landolfo, S. ( 1999; ). Human cytomegalovirus stimulates cellular dihydrofolate reducatse activity in quiescent cells. Intervirology 42, 30–36.[CrossRef]
    [Google Scholar]
  17. Maley, F. & Maley, G. F. ( 1990; ). A tale of two enzymes, deoxycytidylate deaminase and thymidylate synthase. Prog Nucleic Acid Res Mol Biol 39, 49–80.
    [Google Scholar]
  18. Maley, F., Lobo, A. F. & Maley, G. F. ( 1993; ). Properties of an affinity-column purified human deoxycytidylate deaminase. Biochim Biophys Acta 1162, 161–170.[CrossRef]
    [Google Scholar]
  19. McGeoch, D. J. & Davison, A. J. ( 1999; ). The molecular evolutionary history of the herpesviruses. In Origin and Evolution of Viruses, pp. 441–465. Edited by E. Domingo, R. Webster & J. Holland. London: Academic Press.
  20. McIntosh, E. M. & Haynes, R. H. ( 1997; ). dUTP pyrophosphatase as a potential target for chemotherapeutic drug development. Acta Biochim Pol 44, 159–172.
    [Google Scholar]
  21. Mocarski, E. S. & Courcelle, C. T. ( 2001; ). Cytomegalovirus and their replication. In Fields Virology, 4th edn, pp. 2629–2673. Edited by D. M. Knipe & P. M. Howley. Philadelphia: Lippincott Williams & Wilkins.
  22. Pauwels, R., Balzarini, J., Baba, M., Snoeck, R., Schols, D., Hederwijin, P., Desmyter, J. & De Clerq, E. ( 1988; ). Rapid and automated tetrazolium-based colorimetric assay for the detection of anti HIV compounds. J Virol Methods 20, 309–321.[CrossRef]
    [Google Scholar]
  23. Ricceri, G., Ragusa, N., Avola, R., Barcellona, M. L. & Castro, A. ( 1978; ). Pyrimidine nucleotide synthesis in rat embryo cells infected with X14 or H-1 parvovirus. Acta Virol 22, 104–112.
    [Google Scholar]
  24. Rolton, H. A. & Keir, H. M. ( 1974; ). Deoxycytidylate deaminase. Evidence for a new enzyme in cells infected by the virus of herpes simplex. Biochem J 143, 403–409.
    [Google Scholar]
  25. Weiner, K. X. B., Weiner, R. S., Maley, F. & Maley, G. F. ( 1993; ). Primary structure of human deoxycytidylate deaminase and overexpression of its functional protein in Escherichia coli. J Biol Chem 268, 12983–12989.
    [Google Scholar]
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