1887

Abstract

SUMMARY

Aphidicolin, a tetracyclic diterpenoid which inhibits the DNA polymerase-α activities of many eukaryotic cells, inhibited herpes simplex virus growth and DNA synthesis in infected cultures and the activity of the virus DNA polymerase . A wide range of stable aphidicolin sensitivities was represented amongst a collection of virus strains with no prior exposure to this drug, but viruses with polymerase mutations selected for resistance to phosphonoacetic acid (PAA) or to acycloguanosine typically showed increased sensitivity to aphidicolin. Of 16 unrelated PAA-resistant variants, 7 were hypersensitive to aphidicolin. A number of mutants with temperature-sensitive () lesions in the polymerase gene also showed increased aphidicolin sensitivity (e.g. HSV-1[mP17]H) or aphidicolin hypersensitivity (e.g. HSV-1[KOS]D9, C4). Resistance or hypersensitivity of virus growth and DNA synthesis were correlated with resistance or hypersensitivity of virus DNA polymerase reactions . Resistance phenotypes were closely linked to the polymerase gene during recombination with outside markers. Moreover, the selection of aphidicolin-resistant mutants from hypersensitive variants with independent PAA resistance or mutations in the polymerase gene could result in co-selection for PAA-sensitive and phenotypes. Confirmation that multiple independent mutations could determine aphidicolin hypersensitivity was obtained by studies of recombination between independent hypersensitive variants. Aphidicolin-resistant recombinant progeny were formed with recombination frequencies (0.4 to 2.6%) compatible with intragenic events. With parental hypersensitive variants which were products of limited PAA selection, or with the polymerase mutations, aphidicolin-resistant recombinants were PAA-sensitive and/or . The segregation of other markers (, plaque morphology) amongst recombinant progeny permitted the orientation of multiple determinants of PAA resistance and aphidicolin hypersensitivity with respect to other markers in the polymerase gene and in other genes. The nature of residues determined at any one of a constellation of separate sites within the polymerase locus can determine resistance or sensitivity to antiviral drugs and aphidicolin hypersensitivity associated with changes at the polymerase locus facilitates high resolution genetic analysis of this locus.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-65-1-1
1984-01-01
2022-08-18
Loading full text...

Full text loading...

/deliver/fulltext/jgv/65/1/JV0650010001.html?itemId=/content/journal/jgv/10.1099/0022-1317-65-1-1&mimeType=html&fmt=ahah

References

  1. Aron G. M., Purifoy D. J. M., Schaffer P. A. 1975; DNA synthesis and DNA polymerase activity of herpes simplex type 1 temperature-sensitive mutants. Journal of Virology 16:498–507
    [Google Scholar]
  2. Bastow K. F., Derse D. D., Cheng Y.-C. 1983; Susceptibility of phosphonoformic acid-resistant herpes simplex virus variants to arabinosylnucleosides and aphidicolin. Antimicrobial Agents and Chemotherapy 23:914–917
    [Google Scholar]
  3. Bucknall R. A., Moores H., Simms R., Hesp B. 1973; Antiviral effects of aphidicolin, a new antibiotic produced by Cephalosporium aphidicola. Antimicrobial Agents and Chemotherapy 4:294–298
    [Google Scholar]
  4. Chartrand P., Crumpacker C. S., Schaffer P. A., Wilkie N. M. 1980; Physical and genetic analysis of the herpes simplex virus DNA polymerase locus. Virology 103:311–326
    [Google Scholar]
  5. Chartrand P., Stow N. D., Timbury M. C., Wilkie N. M. 1979; Physical mapping of paar mutations of herpes simplex virus type 1 and type 2 by intertypic marker rescue. Journal of Virology 31:265–276
    [Google Scholar]
  6. Coen D. M., Schaffer P. A. 1980; Two distinct loci confer resistance to acycloguanosine in herpes simplex virus type 1. Proceeding of the National Academy of Sciences, U. S. A 77:2265–2269
    [Google Scholar]
  7. Coen D. M., Furman P. A., Gelep P. T., Schaffer P. A. 1982; Mutations in the herpes simplex virus DNA polymerase gene can confer resistance to 9-β-d-arabinofuranosyladenine. Journal of Virology 41:909–918
    [Google Scholar]
  8. Coen D. M., Furman P. A., Aschman D. P., Schaffer P. A. 1983a; Mutations in the herpes simplex virus DNA polymerase gene conferring hypersensitivity to aphidicolin. Nucleic Acids Research 11:5287–5297
    [Google Scholar]
  9. Coen D. M., Aschman D. P., Gelep P. T., Retondo M. J., Weller S. K., Schaffer P. A. 1983b; Fine mapping and molecular cloning of mutations in the herpes simplex virus DNA polymerase locus. Journal of Virology (in press)
    [Google Scholar]
  10. Conley A. J., Knipe D. M., Jones P. C., Roizman B. 1981; Molecular genetics of herpes simplex virus. VII. Characterization of a temperature-sensitive mutant produced by in vitro mutagenesis and defective in DNA synthesis and the accumulation of γ-polypeptides. Journal of Virology 37:191–206
    [Google Scholar]
  11. Crumpacker C. S., Chartrand P., Subak-Sharpe J. H., Wilkie N. M. 1980; Resistance of herpes simplex virus to acycloguanosine - genetic and physical analysis. Virology 105:171–184
    [Google Scholar]
  12. De Luca N., Bzik D. J., Bond V. C., Person S., Snipes W. 1982; Nucleotide sequences of herpes simplex virus type 1 (HSV-1) affecting virus entry, cell fusion and the production of glycoprotein gB (VP7). Virology 122:423
    [Google Scholar]
  13. Derse D., Bastow K. F., Cheng H.-C. 1982; Characterization of the DNA polymerases induced by a group of herpes simplex type 1 variants selected for growth in the presence of phosphonoformic acid. Journal of Biological Chemistry 257:10251–10260
    [Google Scholar]
  14. Dicioccio R. A., Chadha K., Srivastava B. I. S. 1980; Inhibition of herpes simplex virus-induced DNA polymerase, cellular DNA polymerase, and virus production by aphidicolin. Biochimica et biophysica acta 609:224–231
    [Google Scholar]
  15. Dixon R. A. F., Sabourin D. J., Schaffer P. A. 1983; Genetic analysis of temperature-sensitive mutants which define the genes for the major herpes simplex virus type 2 DNA-binding protein and a new late function. Journal of Virology 45:343–353
    [Google Scholar]
  16. Dubbs D. R., Kit S. 1964; Mutant strains of herpes simplex virus deficient in thymidine kinase-inducing activity. Virology 22:493–502
    [Google Scholar]
  17. Erikkson B., Larsson A., Helgstrand E., Iohansson N.-G., Oberg B. 1980; Pyrophosphate analogues as inhibitors of herpes simplex virus type 1 DNA polymerase. Biochimica et biophysica acta 607:53–64
    [Google Scholar]
  18. Field H., Darby G., Wildy P. 1980; Isolation and characterization of acyclovir-resistant mutants of herpes simplex virus. Journal of General Virology 49:115–124
    [Google Scholar]
  19. Furman P. A., Coen D. M., St Clair M. H., Schaffer P. A. 1981; Acyclovir-resistant mutants of herpes simplex virus type 1 express altered DNA polymerase or reduced acyclovir phosphorylating activities. Journal of Virology 40:936–941
    [Google Scholar]
  20. Hay J., Subak-Sharpe J. H. 1976; Mutants of herpes simplex virus types 1 and 2 that are resistant to phosphono-acetic acid induce altered DNA polymerase activities in infected cells. Journal of General Virology 31:145–148
    [Google Scholar]
  21. Hay J., Moss H., Jamieson A. T., Timbury M. C. 1976; Herpes simplex virus proteins: DNA-polymerase and pyrimidine deoxynucleoside kinase activities in temperature-sensitive mutants of herpes simplex virus type 2. Journal of General Virology 31:65–73
    [Google Scholar]
  22. Herrin T. R., Fairgrieve J. S., Bower R. R., Shipkowitz N. L., Mao J. C.-H. 1977; Synthesis and antiherpes activity of analogues of phosphonoacetic acid. Journal of Medicinal Chemistry 20:660–663
    [Google Scholar]
  23. Honess R. W. 1981; Complementation between phosphonoacetic acid-resistant and -sensitive variants of herpes simplex viruses: evidence for an oligomeric protein with restricted intracellular diffusion as the determinant of resistance and sensitivity. Journal of General Virology 57:297–306
    [Google Scholar]
  24. Honess R. W., Watson D. H. 1977; Herpes simplex virus resistance and sensitivity to phosphonoacetic acid. Journal of Virology 21:584–600
    [Google Scholar]
  25. Honess R. W., Buchan A., Halliburton I. W., Watson D. H. 1980; Recombination and linkage between structural and regulatory genes of herpes simplex virus type 1: study of the functional organization of the genome. Journal of Virology 34:716–742
    [Google Scholar]
  26. Huberman J. A. 1981; New views of the biochemistry of eukaryotic DNA replication revealed by aphidicolin, an unusual inhibitor of DNA polymerase. Cell 23:647–648
    [Google Scholar]
  27. Knopf K. W., Kaufman E. R., Crumpacker C. 1981; Physical mapping of drug resistance mutations defines an active centre of the herpes simplex virus DNA polymerase enzyme. Journal of Virology 39:746–757
    [Google Scholar]
  28. Littler E., Purifoy D. J. M., Minson A., Powell K. L. 1983; Herpes simplex virus non-structural proteins. III. Function of the major DNA-binding protein. Journal of General Virology 64:983–995
    [Google Scholar]
  29. Manservigi R., Spear P., Buchan A. 1977; Cell fusion induced by herpes simplex is promoted and suppressed by different viral glycoproteins. Proceedings of the National Academy of Sciences, U. S. A 74:3913–3917
    [Google Scholar]
  30. Mao J. C.-H., Robishaw E. E., Overby L. R. 1975; Inhibition of DNA polymerases from herpes simplex virus-infected Wi-38 cells by phosphonoacetic acid. Journal of Virology 15:1281–1283
    [Google Scholar]
  31. O’Hare P., Honess R. W. 1983; Evidence for a herpesvirus saimiri-specified DNA polymerase activity which is aphidicolin-resistant and phosphonoacetate-sensitive. Journal of General Virology 64:1013–1024
    [Google Scholar]
  32. Pedrali-Noy G., Spadari S. 1979; Effect of aphidicolin on viral and human DNA polymerases. Biochemical and Biophysical Research Communications 88:1194–1202
    [Google Scholar]
  33. Pedrali-Noy G., Spadari S. 1980; Mechanism of inhibition of herpes simplex virus and vaccinia virus DNA polymerases by aphidicolin, a highly specific inhibitor of DNA replication in eukaryotes. Journal of Virology 36:457–464
    [Google Scholar]
  34. Purifoy D. J. M., Powell K. L. 1977; Herpes simplex virus DNA polymerase as the site of phosphonoacetate sensitivity: temperature-sensitive mutants. Journal of Virology 24:470–477
    [Google Scholar]
  35. Purifoy D. J. M., Powell K. L. 1981; Temperature-sensitive mutants in two distinct complementation groups of herpes simplex virus type 1 specify thermolabile DNA polymerase. Journal of General Virology 54:219–222
    [Google Scholar]
  36. Purifoy D. J. M., Lewis R. B., Powell K. L. 1977; Identification of the herpes simplex virus DNA polymerase gene. Nature, London 269:621–623
    [Google Scholar]
  37. Sarmiento M. E., Haffey M., Spear P. G. 1979; Membrane proteins specified by herpes simplex viruses. III. Role of glycoprotein VP7(B2) in virion infectivity. Journal of Virology 29:1149–1158
    [Google Scholar]
  38. Schaffer P. A., Carter V. C., Timbury M. C. 1978; Collaborative complementation study of temperature sensitive mutants of herpes simplex virus types 1 and 2. Journal of Virology 27:490–504
    [Google Scholar]
  39. Schnipper L. E., Crumpacker C. S. 1980; Resistance of herpes simplex virus to acycloguanosine: the role of viral thymidine kinase and DNA polymerase loci. Proceedings of the National Academy of Sciences, U. S. A 77:2270–2273
    [Google Scholar]
  40. Sharp J. A., Wagner M. I., Summers W. 1983; Transcription of herpes simplex virus genes in vivo: overlap of a late promotor with the 3′ end of the early thymidine kinase gene. Journal of Virology 45:10–17
    [Google Scholar]
  41. Smiley J. R., Wagner M. J., Summers W. P., Summers W. C. 1980; Genetic and physical evidence for the polarity of transcription of the thymidine kinase gene of herpes simplex virus. Virology 102:83–93
    [Google Scholar]
  42. Swain M. A., Galloway D. A. 1983; Nucleotide sequence of the herpes simplex virus type 2 thymidine kinase gene. Journal of Virology 46:1045–1050
    [Google Scholar]
  43. Wagner M. J., Sharp J. A., Summers W. C. 1981; Nucleotide sequence of the thymidine kinase gene of herpes simplex virus type 1. Proceedings of the National Academy of Sciences, U. S. A 78:1441–1445
    [Google Scholar]
  44. Weller S. K., Lee K. J., Sabourin D. J., Schaffer P. A. 1983; Genetic analysis of temperature sensitive mutants which define the gene for the major herpes simplex virus type 1 DNA-binding protein. Journal of Virology 45:354–366
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-65-1-1
Loading
/content/journal/jgv/10.1099/0022-1317-65-1-1
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error