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. 1997 Mar;41(3):594–599. doi: 10.1128/aac.41.3.594

Effect of incorporation of cidofovir into DNA by human cytomegalovirus DNA polymerase on DNA elongation.

X Xiong 1, J L Smith 1, M S Chen 1
PMCID: PMC163757  PMID: 9055999

Abstract

Cidofovir (CDV) (HPMPC) has potent in vitro and in vivo activity against human cytomegalovirus (HCMV), CDV diphosphate (CDVpp), the putative antiviral metabolite of CDV, is an inhibitor and an alternate substrate of HCMV DNA polymerase. CDV is incorporated with the correct complementation to dGMP in the template, and the incorporated CDV at the primer end is not excised by the 3'-to-5' exonuclease activity of HCMV DNA polymerase. The incorporation of a CDV molecule causes a decrease in the rate of DNA elongation for the addition of the second natural nucleotide from the singly incorporated CDV molecule. The reduction in the rate of DNA (36-mer) synthesis from an 18-mer by one incorporated CDV is 31% that of the control. However, the fidelity of HCMV DNA polymerase is maintained for the addition of the nucleotides following a single incorporated CDV molecule. The rate of DNA synthesis by HCMV DNA polymerase is drastically decreased after the incorporation of two consecutive CDV molecules; the incorporation of a third consecutive CDV molecule is not detectable. Incorporation of two CDV molecules separated by either one or two deoxynucleoside monophosphates (dAMP, dGMP, or dTMP) also drastically decreases the rate of DNA chain elongation by HCMV DNA polymerase. The rate of DNA synthesis decreases by 90% when a template which contains one internally incorporated CDV molecule is used. The inhibition by CDVpp of DNA synthesis by HCMV DNA polymerase and the inability of HCMV DNA polymerase to excise incorporated CDV from DNA may account for the potent and long-lasting anti-CMV activity of CDV.

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Selected References

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  1. Biron K. K., Stanat S. C., Sorrell J. B., Fyfe J. A., Keller P. M., Lambe C. U., Nelson D. J. Metabolic activation of the nucleoside analog 9-[( 2-hydroxy-1-(hydroxymethyl)ethoxy]methyl)guanine in human diploid fibroblasts infected with human cytomegalovirus. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2473–2477. doi: 10.1073/pnas.82.8.2473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chen M. S., Suttmann R. T., Papp E., Cannon P. D., McRoberts M. J., Bach C., Copeland W. C., Wang T. S. Selective action of 4'-azidothymidine triphosphate on reverse transcriptase of human immunodeficiency virus type 1 and human DNA polymerases alpha and beta. Biochemistry. 1993 Jun 15;32(23):6002–6010. doi: 10.1021/bi00074a011. [DOI] [PubMed] [Google Scholar]
  3. Chen M. S., Suttmann R. T., Wu J. C., Prisbe E. J. Metabolism of 4'-azidothymidine. A compound with potent and selective activity against the human immunodeficiency virus. J Biol Chem. 1992 Jan 5;267(1):257–260. [PubMed] [Google Scholar]
  4. Chen M. S., Walker J., Prusoff W. H. Kinetic studies of herpes simplex virus type 1-encoded thymidine and thymidylate kinase, a multifunctional enzyme. J Biol Chem. 1979 Nov 10;254(21):10747–10753. [PubMed] [Google Scholar]
  5. Cherrington J. M., Allen S. J., McKee B. H., Chen M. S. Kinetic analysis of the interaction between the diphosphate of (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine, ddCTP, AZTTP, and FIAUTP with human DNA polymerases beta and gamma. Biochem Pharmacol. 1994 Nov 16;48(10):1986–1988. doi: 10.1016/0006-2952(94)90600-9. [DOI] [PubMed] [Google Scholar]
  6. Copeland W. C., Chen M. S., Wang T. S. Human DNA polymerases alpha and beta are able to incorporate anti-HIV deoxynucleotides into DNA. J Biol Chem. 1992 Oct 25;267(30):21459–21464. [PubMed] [Google Scholar]
  7. De Clercq E. Broad-spectrum anti-DNA virus and anti-retrovirus activity of phosphonylmethoxyalkylpurines and -pyrimidines. Biochem Pharmacol. 1991 Aug 8;42(5):963–972. doi: 10.1016/0006-2952(91)90276-b. [DOI] [PubMed] [Google Scholar]
  8. Eriksson B., Oberg B., Wahren B. Pyrophosphate analogues as inhibitors of DNA polymerases of cytomegalovirus, herpes simplex virus and cellular origin. Biochim Biophys Acta. 1982 Feb 26;696(2):115–123. doi: 10.1016/0167-4781(82)90018-5. [DOI] [PubMed] [Google Scholar]
  9. Frank K. B., Chiou J. F., Cheng Y. C. Interaction of herpes simplex virus-induced DNA polymerase with 9-(1,3-dihydroxy-2-propoxymethyl)guanine triphosphate. J Biol Chem. 1984 Feb 10;259(3):1566–1569. [PubMed] [Google Scholar]
  10. Freitas V. R., Smee D. F., Chernow M., Boehme R., Matthews T. R. Activity of 9-(1,3-dihydroxy-2-propoxymethyl)guanine compared with that of acyclovir against human, monkey, and rodent cytomegaloviruses. Antimicrob Agents Chemother. 1985 Aug;28(2):240–245. doi: 10.1128/aac.28.2.240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lurain N. S., Spafford L. E., Thompson K. D. Mutation in the UL97 open reading frame of human cytomegalovirus strains resistant to ganciclovir. J Virol. 1994 Jul;68(7):4427–4431. doi: 10.1128/jvi.68.7.4427-4431.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mar E. C., Chiou J. F., Cheng Y. C., Huang E. S. Human cytomegalovirus-induced DNA polymerase and its interaction with the triphosphates of 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-methyluracil, -5-iodocytosine, and -5-methylcytosine. J Virol. 1985 Dec;56(3):846–851. doi: 10.1128/jvi.56.3.846-851.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Reardon J. E. Herpes simplex virus type 1 and human DNA polymerase interactions with 2'-deoxyguanosine 5'-triphosphate analogues. Kinetics of incorporation into DNA and induction of inhibition. J Biol Chem. 1989 Nov 15;264(32):19039–19044. [PubMed] [Google Scholar]
  14. Reid R., Mar E. C., Huang E. S., Topal M. D. Insertion and extension of acyclic, dideoxy, and ara nucleotides by herpesviridae, human alpha and human beta polymerases. A unique inhibition mechanism for 9-(1,3-dihydroxy-2-propoxymethyl)guanine triphosphate. J Biol Chem. 1988 Mar 15;263(8):3898–3904. [PubMed] [Google Scholar]
  15. Xiong X., Smith J. L., Kim C., Huang E. S., Chen M. S. Kinetic analysis of the interaction of cidofovir diphosphate with human cytomegalovirus DNA polymerase. Biochem Pharmacol. 1996 Jun 14;51(11):1563–1567. doi: 10.1016/0006-2952(96)00100-1. [DOI] [PubMed] [Google Scholar]

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