1887

Abstract

Murine cytomegalovirus (MCMV) productively infects quiescent fibroblasts in which the levels of nucleoside triphosphate precursors and cell functions involved in DNA metabolism are minimal. It appears that MCMV has evolved molecular pathways in order to ensure the presence of nucleoside triphosphate precursors for the viral DNA polymerase. Here, we report that MCMV infection of quiescent NIH 3T3 cells markedly stimulates transcription, expression and activity of the cellular dihydrofolate reductase (DHFR), a key enzyme in the synthesis of DNA precursors. DHFR stimulation by MCMV is sensitive to UV irradiation and seems to depend on expression of the viral immediate-early protein pp89. Finally, it has been demonstrated that suppression of virus-induced DHFR activity by the specific inhibitor methotrexate prevents MCMV DNA replication. These observations indicate that induction of host cell DHFR activity by MCMV is required for viral DNA synthesis in quiescent fibroblasts.

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1998-11-01
2022-12-04
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References

  1. Abken H., Reifenrath B. 1992; A procedure to standardize CAT reporter gene assay. Nucleic Acids Research 20:3527
    [Google Scholar]
  2. Allegra C. J., Chabner B. A., Tuazon C. U., Ogata-Arakaki D., Baird B., Drake J. C., Simmons J. T., Lack E. E., Shelhamer J. H., Balis F., Walker R., Kovacs J. A., Lane H. C., Masur H. 1987; Trimetrexate for the treatment of Pneumocystis carinii pneumonia in patients with the acquired immunodeficiency syndrome. New England Journal of Medicine 317:978–985
    [Google Scholar]
  3. Baldanti F., Underwood M. R., Stanat S. C., Biron K., Chou S., Sarasini A., Silini E., Gerna G. 1996; Single amino acid changes in the DNA polymerase confer foscarnet resistance and slow growth phenotype, while mutation in the UL97-encoded phosphotransferase confer ganciclovir resistance in three-double resistant human cytomegalovirus strains recovered from patients with AIDS. Journal of Virology 70:1390–1395
    [Google Scholar]
  4. Bean B. 1992; Antiviral therapy: current concept and practices. Clinical Microbiology Reviews 5:146–182
    [Google Scholar]
  5. Bertino J. R. 1993; Karnofski Memorial Lecture : Ode to methotrexate. Journal of Clinical Oncology 11:5–14
    [Google Scholar]
  6. Boldogh I., AbuBakar S., Millinoff D., Deng C. Z., Albrecht T. 1991; Cellular oncogenes activation by human cytomegalovirus. Lack of correlation with virus infectivity and immediate early gene expression. Archives of Virology 118:163–177
    [Google Scholar]
  7. Burns L. J., Waring F., Reuter J. J., Stinski M. F., Ginder G. 1993; Only the HLA class I minimal promoter elements are required for transactivation by human cytomegalovirus immediate early genes. Blood 81:1558–1566
    [Google Scholar]
  8. Chou S., Erice A., Jordan C. M., Vercellotti G. M., Michelis K. R., Talarico C. L., Stanat S. C., Biron K. 1995; Analysis of UL97 phosphotransferase coding sequence in clinical cytomegalovirus isolates and identification of mutation conferring ganciclovir resistance. Journal of Infectious Disease 171:576–583
    [Google Scholar]
  9. Colberg-Poley A. M., Santomenna L. D., Harlow P. P., Benfield P., Tenney D. J. 1992; Human cytomegalovirus US3 and UL36-38 immediate-early proteins regulate gene expression. Journal of Virology 66:95–105
    [Google Scholar]
  10. Dittmer D., Mocarski E. S. 1997; Human cytomegalovirus infection inhibits G1/S transition. Journal of Virology 71:1629–1634
    [Google Scholar]
  11. Dudding L., Haskill S., Clark B. D., Auron P. E., Sporn S., Huang E. S. 1989; Cytomegalovirus infection stimulates expression of monocyte-associated mediator genes. Journal of Immunology 10:3343–3352
    [Google Scholar]
  12. Fallon J., Allegra C., Kovacs J., O’Neill D., Ogata-Arakaki D., Feuerstein I., Polis M., Davey R., Lane H. C., LaFon S., Rogers M., Zunich M., Zurlo J., Tuazon C., Parenti D., Simon G., Masur H. 1990; Piritrexim with leucovorin for the treatment of pneumocystis pneumonia (PCP) in AIDS patients. Clinical Research 38:361A
    [Google Scholar]
  13. Fleming G. F., Schilsky R. L. 1992; Antifolates : the next generation. Seminars in Oncology 19:707–719
    [Google Scholar]
  14. Furukawa T., Fioretti A., Plotkin S. A. 1973; Growth characteristics of cytomegalovirus in human fibroblasts with demonstration of protein synthesis early in viral replication. Journal of Virology 11:991–997
    [Google Scholar]
  15. Furukawa T., Tanaka S., Plotkin S. A. 1975; Stimulation of macromolecular synthesis in guinea pig cells by human CMV. Proceedings of the Society for Experimental Biology and Medicine 148:211–214
    [Google Scholar]
  16. Gariglio M., Foresta P., Sacchi C., Lembo M., Hertel L., Landolfo S. 1997; Suppression of high mobility group protein T160 expression impairs mouse cytomegalovirus replication. Journal of General Virology 78:665–670
    [Google Scholar]
  17. Geist L. J., Monik M. M., Stinski M. F., Hunninghake G. 1993; The immediate-early genes of human cytomegalovirus upregulate tumor necrosis factor-α gene expression. Journal of Clinical Investigation 93:474–478
    [Google Scholar]
  18. Hayhurst G. P., Bryant L. A., Caswell R. C., Walker S. M., Sinclair J. H. 1995; CCAAT box-dependent activation of the TATA-less human DNA polymerase a promoter by the human cytomegalovirus 72-kilodalton major immediate-early protein. Journal of Virology 69:182–188
    [Google Scholar]
  19. Hitchings G. H. 1978; The metabolism of plasmodia and the chemotherapy of malaria infections. In Tropical Medicine from Romance to Reality p. 79–98 Wood R. J. Edited by London: Academic Press;
    [Google Scholar]
  20. Huang E. S., Kovalik T. F. 1993; The pathogenicity of human cytomegalovirus: an overview. In Molecular Aspects of Human Cyto-megalovirus Diseases pp. 1–45 Becker Y., Darai G., Huang E.-S. Edited by Berlin: Springer-Verlag;
    [Google Scholar]
  21. Johnson L. F., Fuhrman L. G., Weidemann L. M. 1978; Regulation of dihydrofolate reductase gene expression in mouse fibroblasts during the transition from resting to growing state. Journal of Cellular Physiology 97:397–406
    [Google Scholar]
  22. Koszinowski U. H., Keil G. M., Volkmer H., Fibi M. R., Ebeling-Keil A., Munch K. 1986; The 89,000-Mr murine cytomegalovirus immediate-early protein activates gene transcription. Journal of Virology 58:59–66
    [Google Scholar]
  23. Kuyper L. F., Baccanari D. P., Jones M. L., Hunter R. N., Tansik R. L., Joyner S. S., Boytos C. M., Rudolph S. K., Knick V., Wilson H. R., Caddell J. M., Friedman H. S., Comley J. C. W., Stables J. M. 1996; High-affinity inhibitors of dihydrofolate reductase: antimicrobial and anticancer activities of 7,8-dialkyl-1,3-diaminolpyrrolo[3,2-f] quinazolines with small molecular size. Journal of Medicinal Chemistry 39:892–903
    [Google Scholar]
  24. Margolis M. J., Pajovic S., Wong E. L., Wade M., Jupp R., Nelson J. A., Azizkhan J. C. 1995; Interaction of the 72-kilodalton human cytomegalovirus IE1 gene product with E2F1 coincides with E2F-dependent activation of dihydrofolate reductase transcription. Journal of Virology 69:7759–7767
    [Google Scholar]
  25. Nicholas J., Ruvolo V., Zong J., Ciufo D., Guo H. G., Reitz M. S., Hayward G. S. 1997; A single 13-kilobase divergent locus in the Kaposi sarcoma-associated herpesvirus (human herpesvirus 8) genome contains nine open reading frames that are homologous to or related to cellular proteins. Journal of Virology 71:1963–1974
    [Google Scholar]
  26. Rawlinson W. D., Farrell H. E., Barrell B. G. 1996; Analysis of the complete DNA sequence of murine cytomegalovirus. Journal of Virology 70:8833–8849
    [Google Scholar]
  27. Rothenberg S. P. 1966; A rapid radioassay for folic acid reductase and amethopterin. Analytical Biochemistry 16:176–179
    [Google Scholar]
  28. Salter A. J. 1982; Trimethoprim-sulfamethoxazole : an assessment of more than 12 years of use. Review of Infectious Disease 4:196–236
    [Google Scholar]
  29. Schickedanz J., Philipson L., Ansorge W., Pepperkok R., Klein R., Koszinowski U. H. 1988; The 89,000-Mr murine cytomegalovirus immediate-early protein stimulates c-fos expression and cellular DNA synthesis. Journal of Virology 62:3341–3347
    [Google Scholar]
  30. Schweitzer B. I., Dicker A. P., Bertino J. R. 1990; Dihydrofolate reductase as a therapeutic target. FASEB Journal 4:2441–2452
    [Google Scholar]
  31. Shanley J. D., Debs R. J. 1989; The folate antagonist, methotrexate, is a potent inhibitor of murine and human cytomegalovirus in vitro. Antiviral Research 11:99–106
    [Google Scholar]
  32. Tanaka S., Furukawa T., Plotkin S. A. 1975; Human cyto-megalovirus stimulates host cell RNA synthesis. Journal of Virology 15:297–304
    [Google Scholar]
  33. Trimble J. J., Shridhara C. S., Bakker M. A., Grassmann R., Desrosiers R. C. 1988; A gene for dihydrofolate reductase in a herpesvirus. Science 239:1145–1147
    [Google Scholar]
  34. Wachsman M., Hamzeh F. M., Saito H., Lietman P. S. 1996; Anticytomegaloviral activity of methotrexate associated with preferential accumulation of drug by cytomegalovirus-infected cells. Antimicrobial Agents and Chemotherapy 40:433–436
    [Google Scholar]
  35. Wade M., Kowalik T. F., Mudryj M., Huang E. S., Azizkhan J. C. 1992; E2F mediates dihydrofolate reductase promoter activation and multiprotein complex formation in human cytomegalovirus infection. Molecular and Cellular Biology 12:4364–4374
    [Google Scholar]
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