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. 1987 Apr;61(4):1291–1295. doi: 10.1128/jvi.61.4.1291-1295.1987

Isolation and characterization of phosphonoacetic acid-resistant mutants of human cytomegalovirus.

R T D'Aquila, W C Summers
PMCID: PMC254097  PMID: 3029427

Abstract

Mutants of the human cytomegalovirus (HCMV) that were 6- to 13-fold more resistant to phosphonoacetic acid than the wild-type HCMV (Towne) were isolated. Extracts from mycoplasma-free, mutant-infected cells had phosphonoacetate-resistant DNA polymerase activity in vitro. This strongly suggests that the selected mutations are in the HCMV DNA polymerase genes of these viruses.

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Selected References

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  1. Aron G. M., Purifoy D. J., Schaffer P. A. DNA synthesis and DNA polymerase activity of herpes simplex virus type 1 temperature-sensitive mutants. J Virol. 1975 Sep;16(3):498–507. doi: 10.1128/jvi.16.3.498-507.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Chartrand P., Crumpacker C. S., Schaffer P. A., Wilkie N. M. Physical and genetic analysis of the herpes simplex virus DNA polymerase locus. Virology. 1980 Jun;103(2):311–326. doi: 10.1016/0042-6822(80)90190-7. [DOI] [PubMed] [Google Scholar]
  4. Chartrand P., Stow N. D., Timbury M. C., Wilkie N. M. Physical mapping of paar mutations of herpes simplex virus type 1 and type 2 by intertypic marker rescue. J Virol. 1979 Aug;31(2):265–276. doi: 10.1128/jvi.31.2.265-276.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chiou H. C., Weller S. K., Coen D. M. Mutations in the herpes simplex virus major DNA-binding protein gene leading to altered sensitivity to DNA polymerase inhibitors. Virology. 1985 Sep;145(2):213–226. doi: 10.1016/0042-6822(85)90155-2. [DOI] [PubMed] [Google Scholar]
  6. Coen D. M., Aschman D. P., Gelep P. T., Retondo M. J., Weller S. K., Schaffer P. A. Fine mapping and molecular cloning of mutations in the herpes simplex virus DNA polymerase locus. J Virol. 1984 Jan;49(1):236–247. doi: 10.1128/jvi.49.1.236-247.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Coen D. M., Fleming H. E., Jr, Leslie L. K., Retondo M. J. Sensitivity of arabinosyladenine-resistant mutants of herpes simplex virus to other antiviral drugs and mapping of drug hypersensitivity mutations to the DNA polymerase locus. J Virol. 1985 Feb;53(2):477–488. doi: 10.1128/jvi.53.2.477-488.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Coen D. M., Furman P. A., Aschman D. P., Schaffer P. A. Mutations in the herpes simplex virus DNA polymerase gene conferring hypersensitivity to aphidicolin. Nucleic Acids Res. 1983 Aug 11;11(15):5287–5297. doi: 10.1093/nar/11.15.5287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Coen D. M., Furman P. A., Gelep P. T., Schaffer P. A. Mutations in the herpes simplex virus DNA polymerase gene can confer resistance to 9-beta-D-arabinofuranosyladenine. J Virol. 1982 Mar;41(3):909–918. doi: 10.1128/jvi.41.3.909-918.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Crumpacker C. S., Schnipper L. E., Kowalsky P. N., Sherman D. M. Resistance of herpes simplex virus to adenine arabinoside and E-5-(2-bromovinyl)-2'-deoxyuridine: a physical analysis. J Infect Dis. 1982 Aug;146(2):167–172. doi: 10.1093/infdis/146.2.167. [DOI] [PubMed] [Google Scholar]
  11. Demarchi J. M. Human cytomegalovirus DNA: restriction enzyme cleavage maps and map locations for immediate-early, early, and late RNAs. Virology. 1981 Oct 15;114(1):23–38. doi: 10.1016/0042-6822(81)90249-x. [DOI] [PubMed] [Google Scholar]
  12. Derse D., Bastow K. F., Cheng Y. Characterization of the DNA polymerases induced by a group of herpes simplex virus type I variants selected for growth in the presence of phosphonoformic acid. J Biol Chem. 1982 Sep 10;257(17):10251–10260. [PubMed] [Google Scholar]
  13. Dreesman G. R., Benyesh-Melnick M. Spectrum of human cytomegalovirus complement-fixing antigens. J Immunol. 1967 Dec;99(6):1106–1114. [PubMed] [Google Scholar]
  14. Furman P. A., Coen D. M., St Clair M. H., Schaffer P. A. Acyclovir-resistant mutants of herpes simplex virus type 1 express altered DNA polymerase or reduced acyclovir phosphorylating activities. J Virol. 1981 Dec;40(3):936–941. doi: 10.1128/jvi.40.3.936-941.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gibbs J. S., Chiou H. C., Hall J. D., Mount D. W., Retondo M. J., Weller S. K., Coen D. M. Sequence and mapping analyses of the herpes simplex virus DNA polymerase gene predict a C-terminal substrate binding domain. Proc Natl Acad Sci U S A. 1985 Dec;82(23):7969–7973. doi: 10.1073/pnas.82.23.7969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gibson W., Murphy T. L., Roby C. Cytomegalovirus-infected cells contain a DNA-binding protein. Virology. 1981 May;111(1):251–262. doi: 10.1016/0042-6822(81)90669-3. [DOI] [PubMed] [Google Scholar]
  17. Hay J., Moss H., Jamieson A. T., Timbury M. C. Herpesvirus proteins: DNA polymerase and pyrimidine deoxynucleoside kinase activities in temperature-sensitive mutants of herpes simplex virus type 2. J Gen Virol. 1976 Apr;31(1):65–73. doi: 10.1099/0022-1317-31-1-65. [DOI] [PubMed] [Google Scholar]
  18. Hay J., Subak-Sharpe J. H. Mutants of herpes simplex virus types 1 and 2 that are resistant to phosphonoacetic acid induce altered DNA polymerase activities in infected cells. J Gen Virol. 1976 Apr;31(1):145–148. doi: 10.1099/0022-1317-31-1-145. [DOI] [PubMed] [Google Scholar]
  19. Heilbronn R., Jahn G., Bürkle A., Freese U. K., Fleckenstein B., zur Hausen H. Genomic localization, sequence analysis, and transcription of the putative human cytomegalovirus DNA polymerase gene. J Virol. 1987 Jan;61(1):119–124. doi: 10.1128/jvi.61.1.119-124.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hirai K., Furukawa T., Plotkin S. A. Induction of DNA Polymerase in WI-38 and guinea pig cells infected with human cytomegalovirus (HCMV). Virology. 1976 Mar;70(1):251–255. doi: 10.1016/0042-6822(76)90266-x. [DOI] [PubMed] [Google Scholar]
  21. Hirai K., Watanabe Y. Induction of alpha type DNA polymerases in human cytomegalovirus-infected WI-38 cells. Biochim Biophys Acta. 1976 Oct 18;447(3):328–339. doi: 10.1016/0005-2787(76)90056-3. [DOI] [PubMed] [Google Scholar]
  22. Honess R. W., Purifoy D. J., Young D., Gopal R., Cammack N., O'Hare P. Single mutations at many sites within the DNA polymerase locus of herpes simplex viruses can confer hypersensitivity to aphidicolin and resistance to phosphonoacetic acid. J Gen Virol. 1984 Jan;65(Pt 1):1–17. doi: 10.1099/0022-1317-65-1-1. [DOI] [PubMed] [Google Scholar]
  23. Honess R. W., Watson D. H. Herpes simplex virus resistance and sensitivity to phosphonoacetic acid. J Virol. 1977 Feb;21(2):584–600. doi: 10.1128/jvi.21.2.584-600.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Huang E. S. Human cytomegalovirus. III. Virus-induced DNA polymerase. J Virol. 1975 Aug;16(2):298–310. doi: 10.1128/jvi.16.2.298-310.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Huang E. S. Human cytomegalovirus. IV. Specific inhibition of virus-induced DNA polymerase activity and viral DNA replication by phosphonoacetic acid. J Virol. 1975 Dec;16(6):1560–1565. doi: 10.1128/jvi.16.6.1560-1565.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ihara S., Hirai K., Watanabe Y. Temperature-sensitive mutants of human cytomegalovirus: isolation and partial characterization of DNA-minus mutants. Virology. 1978 Jan;84(1):218–221. doi: 10.1016/0042-6822(78)90238-6. [DOI] [PubMed] [Google Scholar]
  27. Jofre J. T., Schaffer P. A., Parris D. S. Genetics of resistance to phosphonoacetic acid in strain KOS of herpes simplex virus type 1. J Virol. 1977 Sep;23(3):833–836. doi: 10.1128/jvi.23.3.833-836.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Knipe D. M., Ruyechan W. T., Roizman B. Molecular genetics of herpes simplex virus. III. Fine mapping of a genetic locus determining resistance to phosphonoacetate by two methods of marker transfer. J Virol. 1979 Feb;29(2):698–704. doi: 10.1128/jvi.29.2.698-704.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Knopf K. W., Kaufman E. R., Crumpacker C. Physical mapping of drug resistance mutations defines an active center of the herpes simplex virus DNA polymerase enzyme. J Virol. 1981 Sep;39(3):746–757. doi: 10.1128/jvi.39.3.746-757.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Leinbach S. S., Reno J. M., Lee L. F., Isbell A. F., Boezi J. A. Mechanism of phosphonoacetate inhibition of herpesvirus-induced DNA polymerase. Biochemistry. 1976 Jan 27;15(2):426–430. doi: 10.1021/bi00647a029. [DOI] [PubMed] [Google Scholar]
  32. Maeda F., Ihara S., Watanabe Y. Morphogenesis of nuclear inclusions and virus capsids in HEL cells infected with temperature-sensitive mutants of human cytomegalovirus. J Gen Virol. 1979 Aug;44(2):419–432. doi: 10.1099/0022-1317-44-2-419. [DOI] [PubMed] [Google Scholar]
  33. Mao J. C., Robishaw E. E. Mode of inhibition of herpes simplex virus DNA polymerase by phosphonoacetate. Biochemistry. 1975 Dec 16;14(25):5475–5479. doi: 10.1021/bi00696a015. [DOI] [PubMed] [Google Scholar]
  34. McDonough S. H., Spector D. H. Transcription in human fibroblasts permissively infected by human cytomegalovirus strain AD169. Virology. 1983 Feb;125(1):31–46. doi: 10.1016/0042-6822(83)90061-2. [DOI] [PubMed] [Google Scholar]
  35. Miller R. L., Rapp F. Distinguishing cytomegalovirus, mycoplasma, and cellular DNA polymerases. J Virol. 1976 Dec;20(3):564–569. doi: 10.1128/jvi.20.3.564-569.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Mocarski E. S., Pereira L., Michael N. Precise localization of genes on large animal virus genomes: use of lambda gt11 and monoclonal antibodies to map the gene for a cytomegalovirus protein family. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1266–1270. doi: 10.1073/pnas.82.4.1266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Nishiyama Y., Maeno K., Yoshida S. Characterization of human cytomegalovirus-induced DNA polymerase and the associated 3'-to-5', exonuclease. Virology. 1983 Jan 30;124(2):221–231. doi: 10.1016/0042-6822(83)90339-2. [DOI] [PubMed] [Google Scholar]
  38. Nowak B., Gmeiner A., Sarnow P., Levine A. J., Fleckenstein B. Physical mapping of human cytomegalovirus genes: identification of DNA sequences coding for a virion phosphoprotein of 71 kDa and a viral 65-kDa polypeptide. Virology. 1984 Apr 15;134(1):91–102. doi: 10.1016/0042-6822(84)90275-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Pande H., Baak S. W., Riggs A. D., Clark B. R., Shively J. E., Zaia J. A. Cloning and physical mapping of a gene fragment coding for a 64-kilodalton major late antigen of human cytomegalovirus. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4965–4969. doi: 10.1073/pnas.81.15.4965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Purifoy D. J., Benyesh-Melnick M. DNA polymerase induction by DNA-negative temperature-sensitive mutants of herpes simplex virus type 2. Virology. 1975 Dec;68(2):374–386. doi: 10.1016/0042-6822(75)90280-9. [DOI] [PubMed] [Google Scholar]
  41. Purifoy D. J., Lewis R. B., Powell K. L. Identification of the herpes simplex virus DNA polymerase gene. Nature. 1977 Oct 13;269(5629):621–623. doi: 10.1038/269621a0. [DOI] [PubMed] [Google Scholar]
  42. Purifoy D. J., Powell K. L. Herpes simplex virus DNA polymerase as the site of phosphonoacetate sensitivity: temperature-sensitive mutants. J Virol. 1977 Nov;24(2):470–477. doi: 10.1128/jvi.24.2.470-477.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Russell W. C., Newman C., Williamson D. H. A simple cytochemical technique for demonstration of DNA in cells infected with mycoplasmas and viruses. Nature. 1975 Feb 6;253(5491):461–462. doi: 10.1038/253461a0. [DOI] [PubMed] [Google Scholar]
  44. Schneider E. L., Stanbridge E. J. Comparison of methods for the detection of Mycoplasmal contamination of cell cultures: a review. In Vitro. 1975 Jan-Feb;11(1):20–34. doi: 10.1007/BF02615318. [DOI] [PubMed] [Google Scholar]
  45. Schneider E. L., Stanbridge E. J., Epstein C. J. Incorporation of 3H-uridine and 3H-uracil into RNA: a simple technique for the detection of mycoplasma contamination of cultured cells. Exp Cell Res. 1974 Mar 15;84(1):311–318. doi: 10.1016/0014-4827(74)90411-x. [DOI] [PubMed] [Google Scholar]
  46. Schreier P. H., Cortese R. A fast and simple method for sequencing DNA cloned in the single-stranded bacteriophage M13. J Mol Biol. 1979 Mar 25;129(1):169–172. doi: 10.1016/0022-2836(79)90068-8. [DOI] [PubMed] [Google Scholar]
  47. Stenberg R. M., Thomsen D. R., Stinski M. F. Structural analysis of the major immediate early gene of human cytomegalovirus. J Virol. 1984 Jan;49(1):190–199. doi: 10.1128/jvi.49.1.190-199.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Stinski M. F., Thomsen D. R., Stenberg R. M., Goldstein L. C. Organization and expression of the immediate early genes of human cytomegalovirus. J Virol. 1983 Apr;46(1):1–14. doi: 10.1128/jvi.46.1.1-14.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Suzuki S., Kimura T., Saneyoshi M. Characterization of DNA polymerase induced by salmon herpesvirus, Oncorhynchus masou virus. J Gen Virol. 1986 Feb;67(Pt 2):405–408. doi: 10.1099/0022-1317-67-2-405. [DOI] [PubMed] [Google Scholar]
  50. Yamanishi K., Rapp F. Induction of host DNA synthesis and DNA polymerase by DNA-negative temperature-sensitive mutants of human cytomegalovirus. Virology. 1979 Apr 15;94(1):237–241. doi: 10.1016/0042-6822(79)90457-4. [DOI] [PubMed] [Google Scholar]
  51. Yamanishi K., Rapp F. Temperature-sensitive mutants of human cytomegalovirus. J Virol. 1977 Oct;24(1):416–418. doi: 10.1128/jvi.24.1.416-418.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Yoshida S., Kondo T., Ando T. Multiple molecular species of cytoplasmic DNA polymerase from calf thymus. Biochim Biophys Acta. 1974 Jul 24;353(4):463–474. doi: 10.1016/0005-2787(74)90052-5. [DOI] [PubMed] [Google Scholar]

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