1887

Journal of General Virology header logo

Volume 66, Issue 1

Studies on the Herpes Simplex Virus Alkaline Nuclease: Detection of Type-common and Type-specific Epitopes on the Enzyme Free

Abstract

SUMMARY

Five monoclonal antibodies to the alkaline nuclease of herpes simplex virus (HSV) types 1 and 2 have been used in immunoperoxidase tests to demonstrate the nuclear localization of the enzyme within HSV-1- and HSV-2-infected cells and to purify the enzyme from cells infected with either virus by immunoadsorbant chromatography. Affinity chromatography with a P-labelled extract of HSV-2-infected cells has enabled us to demonstrate that the nuclease eluting from the immunoadsorbant is a phosphoprotein, hence confirming the nuclease to be identical to the phosphorylated polypeptide previously referred to as ICSP 22 (HSV-2) or ICP 19 (HSV-1). In addition, the results clearly demonstrate that monoclonal antibodies Q1, CC1 and CH2 are directed against HSV type-common epitopes while V1 and TT1 antibodies are against HSV-2-specific epitopes on the enzyme. Using the type-specific monoclonal antibodies in an immunoperoxidase test, the enzyme specified in cells infected with intertypic recombinants has been typed; correlation of these data with restriction endonuclease maps of the recombinants has enabled us to map the position of the active site of the nuclease gene to map units 0.168 to 0.184 on the genomes of both HSV-1 and HSV-2. Taken with the data mapping the mRNA encoding this enzyme, the nuclease active site can be mapped to 0.168 to 0.175 on the genome. Finally, the use of monoclonal antibodies in immunofluorescence tests on infected cells has demonstrated that the nuclease is synthesized within 2 h post-infection.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): 2 , alkaline , epitopes , HSV types 1 and nuclease
© Journal of General Virology 1985
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-66-1-1
1985-01-01
2024-04-23
Loading full text...

Full text loading...

/deliver/fulltext/jgv/66/1/JV0660010001.html?itemId=/content/journal/jgv/10.1099/0022-1317-66-1-1&mimeType=html&fmt=ahah

References

  1. Banks L., Purifoy D. J. M., Hurst P-F., Killington R. A., Powell K. L. 1953; Herpes simplex virus nonstructural proteins. IV. Purification of the virus-induced deoxyribonuclease and characterization of the enzyme using monoclonal antibodies. Journal of General Virology 64:2249–2260
     [Google Scholar]
  2. Banks L. M., Vaughan P. J., Meredith D., Powell K. L. 1984; Monoclonal antibodies to herpes simplex virus thymidine kinase. Journal of General Virology 65:1625–1630
     [Google Scholar]
  3. Bonner W. M., Laskey R. A. 1974; A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46:83–88
     [Google Scholar]
  4. Costa R. H., Draper K. G., Banks L. M., Powell K. L., Cohen G., Eisenberg R., Wagner E. K. 1983; High-resolution characterisation of herpes simplex virus type 1 transcripts encoding alkaline nuclease and a 50,000-dalton protein tentatively identified as capsid protein. Journal of Virology 48:591–603
     [Google Scholar]
  5. Fahey J. L., Horbett A. P. 1959; Human gamma globulin fractionation on anion exchange cellulose columns. Journal of Biological Chemistry 234:2645–2651
     [Google Scholar]
  6. Francke B., Garrett B. 1982; The effect of a temperature-sensitive lesion in the alkaline DNAse of herpes simplex virus type 2 on the synthesis of viral DNA. Virology 116:116–127
     [Google Scholar]
  7. Halliburton I. W. 1980; Intertypic recombinants of herpes simplex viruses. Journal of General Virology 48:1–23
     [Google Scholar]
  8. Halliburton I. W., Randall R. E., Killington R. A., Watson D. H. 1977; Some properties of recombinants between type 1 and type 2 herpes simplex viruses. Journal of General Virology 36:471–484
     [Google Scholar]
  9. Hoffman P. J., Cheng Y. C. 1978; The deoxyribonuclease induced after infection of KB cells by herpes simplex virus type 1 or type 2.I. Purification and characterisation of the enzyme. Journal of Biological Chemistry 253:3557–3562
     [Google Scholar]
  10. Hoffman P. J., Cheng Y. C. 1979; DNAse induced after infection of KB cells by herpes simplex virus type 1 or type 2. II. Characterisation of an associated endonuclease activity. Journal of Virology 32:449–457
     [Google Scholar]
  11. Hoggan H. D., Roizman B. 1959; The isolation and properties of a variant of herpes simplex virus producing multinucleated giant cells in monolayer cultures in the presence of antibody. American Journal of Hygiene 70:208–219
     [Google Scholar]
  12. Keir H. M., Gold E. 1963; Deoxyribonucleic acid nucleotidyl transferase and deoxyribonuclease from cultured cells infected with herpes simplex virus. Biochimica et biophysica acta 72:263–276
     [Google Scholar]
  13. Marsden H. S., Stow N. D., Preston V. G., Timbury M. C., Wilkie N. M. 1978; Phyical mapping of herpes simplex virus induced polypeptides. Journal of Virology 28:624–642
     [Google Scholar]
  14. Morse L. S., Pereira L., Roizman B., Schaffer P. A. 1978; Anatomy of herpes simplex virus DNA. X. Mapping of viral genes by analysis of polypeptides and functions specified by HSV-1 × HSV-2 recombinants. Journal of Virology 26:389–410
     [Google Scholar]
  15. Pereira L., Wolff M. H., Fenwick M., Roizman B. 1977; Regulation of herpes virus macromolecular synthesis. V. Properties of two polypeptides made in HSV-1 and HSV-2 infected cells. Virology 77:733–749
     [Google Scholar]
  16. Powell K. L., Courtney R. J. 1975; Polypeptides synthesized in herpes simplex virus infected HEp-2 cells. Virology 66:217–228
     [Google Scholar]
  17. Powell K. L., Purifoy D. J. M. 1977; Nonstructural proteins of herpes simplex virus. I. Purification of the induced DNA polymerase. Journal of Virology 24:618–626
     [Google Scholar]
  18. Powell K. L., Mirkovic R., Courtney R. J. 1977; Comparative analysis of polypeptides induced by type 1 and type 2 strains of herpes simplex virus. Intervirology 8:18–29
     [Google Scholar]
  19. Preston C. M., Cordingley M. G. 1982; mRNA- and DNA-directed synthesis of herpes simplex virus-coded exonuclease in Xenopus laevis oocytes. Journal of Virology 43:386–394
     [Google Scholar]
  20. Purifoy D. J. M., Benyesh-Melnick M. 1975; DNA polymerase induction by DNA-negative temperature-sensitive mutants of herpes simplex virus type 2. Virology 68:374–386
     [Google Scholar]
  21. Purifoy D. J. M., Powell K. L. 1976; DNA binding proteins induced by herpes simplex virus in HEp-2 cells. Journal of Virology 19:717–731
     [Google Scholar]
  22. Rawls W. E., Laurel D., Melnick J. L., Glicksman J. M., Kaufman R. H. 1968; A search for viruses in smegma, premalignant and early malignant cervical tissues. The isolation of herpesviruses with distinct antigenic properties. American Journal of Epidemiology 87:647–655
     [Google Scholar]
  23. Sim C., Watson D. H. 1973; The role of type specific and cross reacting structural antigens in the neutralization of herpes simplex virus types 1 and 2. Journal of General Virology 19:217–233
     [Google Scholar]
  24. Smith K. O. 1964; Relationship between the envelope and the infectivity of herpes simplex virus. Proceedings of the Society for Experimental Biology and Medicine 115:814–816
     [Google Scholar]
  25. Strobel-Fidler M., Francke B. 1980; Alkaline deoxyribonuclease induced by herpes simplex virus type 1: composition and properties of the purified enzyme. Virology 103:493–501
     [Google Scholar]
  26. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, U.S.A 76:4350–4354
     [Google Scholar]
  27. Vaughan P. J., Banks L. M., Purifoy D. J. M., Powell K. L. 1984; Interactions between herpes simplex virus DNA-binding proteins. Journal of General Virology 65:2033–2041
     [Google Scholar]
  28. Watson D. H., Shedden W. I. H., Elliot A., Tetsuka T., Wildy P., Bourgaux-Ramoisy D., Gold E. 1966; Virus specific antigens in mammalian cells infected with herpes simplex virus. Immunology 11:399–408
     [Google Scholar]
  29. Wilcox K. W., Kohn A., Sklyanskaya E., Roizman B. 1980; Herpes simplex virus phosphoproteins. I. Phosphate cycles on and off some viral polypeptides and can alter their affinity for DNA. Journal of Virology 33:167–182
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-66-1-1
Loading
Studies on the Herpes Simplex Virus Alkaline Nuclease: Detection of Type-common and Type-specific Epitopes on the Enzyme
J. Gen. Virol. 66, 1 (1985); https://doi.org/10.1099/0022-1317-66-1-1
/content/journal/jgv/10.1099/0022-1317-66-1-1
/content/journal/jgv/10.1099/0022-1317-66-1-1
Loading

Data & Media loading...

Most cited 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