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. 1983 Jul;24(1):10–14. doi: 10.1128/aac.24.1.10

Phosphorylation of acyclovir in vitro in activated Burkitt somatic cell hybrids.

A K Datta, J S Pagano
PMCID: PMC185096  PMID: 6312870

Abstract

Acyclovir [9-(2-hydroxyethoxymethyl)guanine] (ACV), a potent antiviral compound, was phosphorylated to the same extent by extracts from untreated and iododeoxyuridine-treated Epstein-Barr virus-containing latent D98/HR-1 somatic hybrid cells. ATP was the preferred phosphate donor over other nucleoside triphosphates. The cytosol extract from D98/HR-1 cells effected optimum phosphorylation of thymidine at pH 8.0, whereas ACV was phosphorylated equally well over a wide pH range. Electrophoretic analysis of thymidine kinase-, deoxycytidine kinase-, and ACV-phosphorylating activities from both untreated and iododeoxyuridine-treated cell extracts displayed identical properties. A small part (5 to 10%) of the loaded ACV-phosphorylating activity seemed to migrate with the deoxycytidine kinase activity from cytosol. dTTP and dCTP, at relatively high concentrations, partially inhibited ACV-phosphorylating activity. The results suggest that Epstein-Barr virus does not code for its own thymidine kinase and that phosphorylation of ACV in Epstein-Barr virus-producing cells is carried out by multiple or as yet unidentified ATP-dependent nonspecific cellular phosphotransferases.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Brockman R. W., Cheng Y. C., Schabel F. M., Jr, Montgomery J. A. Metabolism and chemotherapeutic activity of 9-beta-D-arabinofuranosyl-2-fluoroadenine against murine leukemia L1210 and evidence for its phosphorylation by deoxycytidine kinase. Cancer Res. 1980 Oct;40(10):3610–3615. [PubMed] [Google Scholar]
  2. Burns W. H., Wingard J. R., Bender W. J., Saral R. Thymidine kinase not required for antiviral activity of acyclovir against mouse cytomegalovirus. J Virol. 1981 Sep;39(3):889–893. doi: 10.1128/jvi.39.3.889-893.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chen S. T., Estes J. E., Huang E. S., Pagano J. S. Epstein-Barr virus-associated thymidine kinase. J Virol. 1978 Apr;26(1):203–208. doi: 10.1128/jvi.26.1.203-208.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Coen D. M., Schaffer P. A. Two distinct loci confer resistance to acycloguanosine in herpes simplex virus type 1. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2265–2269. doi: 10.1073/pnas.77.4.2265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Colby B. M., Furman P. A., Shaw J. E., Elion G. B., Pagano J. S. Phosphorylation of acyclovir [9-(2-hydroxyethoxymethyl)guanine] in Epstein-Barr virus-infected lymphoblastoid cell lines. J Virol. 1981 May;38(2):606–611. doi: 10.1128/jvi.38.2.606-611.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Colby B. M., Shaw J. E., Elion G. B., Pagano J. S. Effect of acyclovir [9-(2-hydroxyethoxymethyl)guanine] on Epstein-Barr virus DNA replication. J Virol. 1980 May;34(2):560–568. doi: 10.1128/jvi.34.2.560-568.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Datta A. K., Colby B. M., Shaw J. E., Pagano J. S. Acyclovir inhibition of Epstein-Barr virus replication. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5163–5166. doi: 10.1073/pnas.77.9.5163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Datta A. K., Feighny R. J., Pagano J. S. Induction of Epstein-Barr virus-associated DNA polymerase by 12-O-tetradecanoylphorbol-13-acetate. Purification and characterization. J Biol Chem. 1980 Jun 10;255(11):5120–5125. [PubMed] [Google Scholar]
  9. Davidson R. L., Kaufman E. R., Crumpacker C. S., Schnipper L. E. Inhibition of herpes simplex virus transformed and nontransformed cells by acycloguanosine: mechanisms of uptake and toxicity. Virology. 1981 Aug;113(1):9–19. doi: 10.1016/0042-6822(81)90132-x. [DOI] [PubMed] [Google Scholar]
  10. Elion G. B., Furman P. A., Fyfe J. A., de Miranda P., Beauchamp L., Schaeffer H. J. Selectivity of action of an antiherpetic agent, 9-(2-hydroxyethoxymethyl) guanine. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5716–5720. doi: 10.1073/pnas.74.12.5716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Glaser R., Nonoyama M., Decker B., Rapp F. Synthesis of Epstein-Barr virus antigens and DNA in activated Burkitt somatic cell hybrids. Virology. 1973 Sep;55(1):62–69. doi: 10.1016/s0042-6822(73)81008-6. [DOI] [PubMed] [Google Scholar]
  12. Honess R. W., O'Hare P., Young D. Comparison of thymidine kinase activities indiced in cells productively infected with herpesvirus saimiri and herpes simplex virus. J Gen Virol. 1982 Feb;58(Pt 2):237–249. doi: 10.1099/0022-1317-58-2-237. [DOI] [PubMed] [Google Scholar]
  13. Ives D. H., Durham J. P. Deoxycytidine kinase. 3. Kinetics and allosteric regulation of the calf thymus enzyme. J Biol Chem. 1970 May 10;245(9):2285–2294. [PubMed] [Google Scholar]
  14. Kit S., Leung W. C., Trkula D. Properties of mitochondrial thymidine kinases of parental and enzyme-deficient HeLa cells. Arch Biochem Biophys. 1973 Oct;158(2):503–513. doi: 10.1016/0003-9861(73)90542-0. [DOI] [PubMed] [Google Scholar]
  15. Leung W. C., Dubbs D. R., Trkula D., Kit S. Mitochondrial and herpesvirus-specific deoxypyrimidine kinases. J Virol. 1975 Sep;16(3):486–497. doi: 10.1128/jvi.16.3.486-497.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Miller R. L., Glaser R., Rapp F. Studies of an Epstein-Barr virus-induced DNA polymerase. Virology. 1977 Feb;76(2):494–502. doi: 10.1016/0042-6822(77)90232-x. [DOI] [PubMed] [Google Scholar]
  17. Miller W. H., Miller R. L. Phosphorylation of acyclovir (acycloguanosine) monophosphate by GMP kinase. J Biol Chem. 1980 Aug 10;255(15):7204–7207. [PubMed] [Google Scholar]
  18. Pagano J. S., Datta A. K. Perspectives on interactions of acyclovir with Epstein-Barr and other herpes viruses. Am J Med. 1982 Jul 20;73(1A):18–26. doi: 10.1016/0002-9343(82)90057-2. [DOI] [PubMed] [Google Scholar]
  19. Roubal J., Klein G. Synthesis of thymidine kinase (TK) in Epstein-Barr virus-superinfected Raji TK-negative cells. Intervirology. 1981;15(1):43–48. doi: 10.1159/000149213. [DOI] [PubMed] [Google Scholar]

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