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. 1995 Feb;69(2):796–800. doi: 10.1128/jvi.69.2.796-800.1995

A three-nucleotide deletion in the UL97 open reading frame is responsible for the ganciclovir resistance of a human cytomegalovirus clinical isolate.

F Baldanti 1, E Silini 1, A Sarasini 1, C L Talarico 1, S C Stanat 1, K K Biron 1, M Furione 1, F Bono 1, G Palù 1, G Gerna 1
PMCID: PMC188644  PMID: 7815545

Abstract

Multiple human cytomegalovirus (HCMV) strains frequently coexist in patients with AIDS, and chronic ganciclovir treatment may favor the emergence of ganciclovir-resistant viral mutants. We report the molecular and biochemical characterization of a HCMV ganciclovir-resistant strain (VR3480) previously recovered from a patient with AIDS who was undergoing multiple courses of ganciclovir treatment (G. Gerna, F. Baldanti, M. Zavattoni, A. Sarasini, E. Percivalle, and M. G. Revello, Antiviral Res. 19:333-345, 1992). Ganciclovir resistance of strain VR3480 was related to impaired ability to monophosphorylate the drug, as indicated by the finding that ganciclovir phosphorylation values for VR3480 were 30% of those shown by the HCMV reference strain AD169 in an in vitro activity assay. Sequencing of the UL97 gene of VR3480, which encodes the viral kinase responsible for ganciclovir phosphorylation, showed an in-frame deletion of three nucleotides resulting in the loss of a leucine at position 595 in the polypeptide. Mutant VR3480 UL97 DNA was able to transfer resistance to the AD169 strain in marker rescue experiments. Analysis of virus isolates and blood polymorphonuclear leukocyte samples spanning the 2-year follow-up period of the patient showed that ganciclovir-resistant strain VR3480 arose ex novo during prolonged antiviral treatment and accounted for the majority of virus load circulating in blood during the period of clinical resistance to ganciclovir treatment.

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Selected References

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  1. Biron K. K., Fyfe J. A., Stanat S. C., Leslie L. K., Sorrell J. B., Lambe C. U., Coen D. M. A human cytomegalovirus mutant resistant to the nucleoside analog 9-([2-hydroxy-1-(hydroxymethyl)ethoxy]methyl)guanine (BW B759U) induces reduced levels of BW B759U triphosphate. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8769–8773. doi: 10.1073/pnas.83.22.8769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chee M. S., Lawrence G. L., Barrell B. G. Alpha-, beta- and gammaherpesviruses encode a putative phosphotransferase. J Gen Virol. 1989 May;70(Pt 5):1151–1160. doi: 10.1099/0022-1317-70-5-1151. [DOI] [PubMed] [Google Scholar]
  3. Dankner W. M., Scholl D., Stanat S. C., Martin M., Sonke R. L., Spector S. A. Rapid antiviral DNA-DNA hybridization assay for human cytomegalovirus. J Virol Methods. 1990 Jun;28(3):293–298. doi: 10.1016/0166-0934(90)90122-v. [DOI] [PubMed] [Google Scholar]
  4. Drew W. L., Miner R. C., Busch D. F., Follansbee S. E., Gullett J., Mehalko S. G., Gordon S. M., Owen W. F., Jr, Matthews T. R., Buhles W. C. Prevalence of resistance in patients receiving ganciclovir for serious cytomegalovirus infection. J Infect Dis. 1991 Apr;163(4):716–719. doi: 10.1093/infdis/163.4.716. [DOI] [PubMed] [Google Scholar]
  5. Erice A., Chou S., Biron K. K., Stanat S. C., Balfour H. H., Jr, Jordan M. C. Progressive disease due to ganciclovir-resistant cytomegalovirus in immunocompromised patients. N Engl J Med. 1989 Feb 2;320(5):289–293. doi: 10.1056/NEJM198902023200505. [DOI] [PubMed] [Google Scholar]
  6. Fleckenstein B., Müller I., Collins J. Cloning of the complete human cytomegalovirus genome in cosmids. Gene. 1982 Apr;18(1):39–46. doi: 10.1016/0378-1119(82)90054-3. [DOI] [PubMed] [Google Scholar]
  7. Fyfe J. A., McKee S. A., Keller P. M. Altered thymidine-thymidylate kinases from strains of herpes simplex virus with modified drug sensitivities to acyclovir and (E)-5-(2-bromovinyl)-2'-deoxyuridine. Mol Pharmacol. 1983 Sep;24(2):316–323. [PubMed] [Google Scholar]
  8. Gerna G., Baldanti F., Zavattoni M., Sarasini A., Percivalle E., Revello M. G. Monitoring of ganciclovir sensitivity of multiple human cytomegalovirus strains coinfecting blood of an AIDS patient by an immediate-early antigen plaque assay. Antiviral Res. 1992 Oct 1;19(4):333–345. doi: 10.1016/0166-3542(92)90014-v. [DOI] [PubMed] [Google Scholar]
  9. Gerna G., Parea M., Percivalle E., Zipeto D., Silini E., Barbarini G., Milanesi G. Human cytomegalovirus viraemia in HIV-1-seropositive patients at various clinical stages of infection. AIDS. 1990 Oct;4(10):1027–1031. doi: 10.1097/00002030-199010000-00014. [DOI] [PubMed] [Google Scholar]
  10. Knighton D. R., Zheng J. H., Ten Eyck L. F., Xuong N. H., Taylor S. S., Sowadski J. M. Structure of a peptide inhibitor bound to the catalytic subunit of cyclic adenosine monophosphate-dependent protein kinase. Science. 1991 Jul 26;253(5018):414–420. doi: 10.1126/science.1862343. [DOI] [PubMed] [Google Scholar]
  11. Kouzarides T., Bankier A. T., Satchwell S. C., Weston K., Tomlinson P., Barrell B. G. Sequence and transcription analysis of the human cytomegalovirus DNA polymerase gene. J Virol. 1987 Jan;61(1):125–133. doi: 10.1128/jvi.61.1.125-133.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Littler E., Stuart A. D., Chee M. S. Human cytomegalovirus UL97 open reading frame encodes a protein that phosphorylates the antiviral nucleoside analogue ganciclovir. Nature. 1992 Jul 9;358(6382):160–162. doi: 10.1038/358160a0. [DOI] [PubMed] [Google Scholar]
  13. Lurain N. S., Thompson K. D., Holmes E. W., Read G. S. Point mutations in the DNA polymerase gene of human cytomegalovirus that result in resistance to antiviral agents. J Virol. 1992 Dec;66(12):7146–7152. doi: 10.1128/jvi.66.12.7146-7152.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Orita M., Suzuki Y., Sekiya T., Hayashi K. Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics. 1989 Nov;5(4):874–879. doi: 10.1016/0888-7543(89)90129-8. [DOI] [PubMed] [Google Scholar]
  15. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Stanat S. C., Reardon J. E., Erice A., Jordan M. C., Drew W. L., Biron K. K. Ganciclovir-resistant cytomegalovirus clinical isolates: mode of resistance to ganciclovir. Antimicrob Agents Chemother. 1991 Nov;35(11):2191–2197. doi: 10.1128/aac.35.11.2191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sullivan V., Biron K. K., Talarico C., Stanat S. C., Davis M., Pozzi L. M., Coen D. M. A point mutation in the human cytomegalovirus DNA polymerase gene confers resistance to ganciclovir and phosphonylmethoxyalkyl derivatives. Antimicrob Agents Chemother. 1993 Jan;37(1):19–25. doi: 10.1128/aac.37.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sullivan V., Talarico C. L., Stanat S. C., Davis M., Coen D. M., Biron K. K. A protein kinase homologue controls phosphorylation of ganciclovir in human cytomegalovirus-infected cells. Nature. 1992 Jul 9;358(6382):162–164. doi: 10.1038/358162a0. [DOI] [PubMed] [Google Scholar]
  19. Wang T. S., Wong S. W., Korn D. Human DNA polymerase alpha: predicted functional domains and relationships with viral DNA polymerases. FASEB J. 1989 Jan;3(1):14–21. doi: 10.1096/fasebj.3.1.2642867. [DOI] [PubMed] [Google Scholar]
  20. Zipeto D., Revello M. G., Silini E., Parea M., Percivalle E., Zavattoni M., Milanesi G., Gerna G. Development and clinical significance of a diagnostic assay based on the polymerase chain reaction for detection of human cytomegalovirus DNA in blood samples from immunocompromised patients. J Clin Microbiol. 1992 Feb;30(2):527–530. doi: 10.1128/jcm.30.2.527-530.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

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