Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1977 Nov;24(2):618–626. doi: 10.1128/jvi.24.2.618-626.1977

Nonstructural proteins of herpes simplex virus. I. Purification of the induced DNA polymerase.

K L Powell, D J Purifoy
PMCID: PMC515973  PMID: 21304

Abstract

Herpes simplex virus-induced DNA polymerase purified by published methods was found to be contaminated with many others proteins, including virus structural proteins. Thus, DEAE-cellulose and phosphocellulose chromatography were used in combination with affinity chromatography to purify DNA polymerase from herpes simplex virus type 1- and type 2-infected cells. The purified enzyme retained unique features of the herpesvirus-induced DNA polymerase, including a requirement for high salt concentrations for maximal activity, a sensitivity to low phosphonoacetate concentrations, and the capacity to be neutralized by rabbit antiserum to herpesvirus-infected cells. By polyacrylamide gel electrophoresis, the purified DNA polymerase was associated with a virus-induced polypeptide of about 150,000 molecular weight.

Full text

PDF
618

Images in this article

623Image
on p.623

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allen G. P., O'Callaghan D. J., Randall C. C. Purification and characterization of equine herpesvirus-induced DNA. Virology. 1977 Jan;76(1):395–408. doi: 10.1016/0042-6822(77)90311-7. [DOI] [PubMed] [Google Scholar]
  2. FAHEY J. L., HORBETT A. P. Human gamma globulin fractionation on anion exchange cellulose columns. J Biol Chem. 1959 Oct;234:2645–2651. [PubMed] [Google Scholar]
  3. Honess R. W., Roizman B. Proteins specified by herpes simplex virus. XI. Identification and relative molar rates of synthesis of structural and nonstructural herpes virus polypeptides in the infected cell. J Virol. 1973 Dec;12(6):1347–1365. doi: 10.1128/jvi.12.6.1347-1365.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Honess R. W., Roizman B. Regulation of herpesvirus macromolecular synthesis. I. Cascade regulation of the synthesis of three groups of viral proteins. J Virol. 1974 Jul;14(1):8–19. doi: 10.1128/jvi.14.1.8-19.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Honess R. W., Roizman B. Regulation of herpesvirus macromolecular synthesis: sequential transition of polypeptide synthesis requires functional viral polypeptides. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1276–1280. doi: 10.1073/pnas.72.4.1276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Honess R. W., Watson D. H. Herpes simplex virus-specific polypeptides studied by polyacrylamide gel electrophoresis of immune precipitates. J Gen Virol. 1974 Feb;22(2):171–185. doi: 10.1099/0022-1317-22-2-171. [DOI] [PubMed] [Google Scholar]
  8. KIT S., DUBBS D. R. Acquisition of thymidine kinase activity by herpes simplex-infected mouse fibroblast cells. Biochem Biophys Res Commun. 1963 Apr 2;11:55–59. doi: 10.1016/0006-291x(63)90027-5. [DOI] [PubMed] [Google Scholar]
  9. Keir H. M., Hay J., Morrison J. M., Subak-Sharpe H. Altered properties of deoxyribonucleic acid nucleotidyltransferase after infection of mammalian cells with herpes simplex virus. Nature. 1966 Apr 23;210(5034):369–371. doi: 10.1038/210369a0. [DOI] [PubMed] [Google Scholar]
  10. Keir H. M., Subak-Sharpe H., Shedden W. I., Watson D. H., Wildy P. Immunological evidence for a specific DNA polymerase produced after infection by herpes simplex virus. Virology. 1966 Sep;30(1):154–157. doi: 10.1016/s0042-6822(66)81022-x. [DOI] [PubMed] [Google Scholar]
  11. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  12. Mao J. C., Robishaw E. E., Overby L. R. Inhibition of DNA polymerase from herpes simplex virus-infected wi-38 cells by phosphonoacetic Acid. J Virol. 1975 May;15(5):1281–1283. doi: 10.1128/jvi.15.5.1281-1283.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Pereira L., Wolff M. H., Fenwick M., Roizman B. Regulation of herpesvirus macromolecular synthesis. V. Properties of alpha polypeptides made in HSV-1 and HSV-2 infected cells. Virology. 1977 Apr;77(2):733–749. doi: 10.1016/0042-6822(77)90495-0. [DOI] [PubMed] [Google Scholar]
  14. Powell K. L., Courtney R. J. Polypeptide synthesized in herpes simplex virus type 2-infected HEp-2 cells. Virology. 1975 Jul;66(1):217–228. doi: 10.1016/0042-6822(75)90192-0. [DOI] [PubMed] [Google Scholar]
  15. Powell K. L., Mirkovic R., Courtney R. J. Comparative analysis of polypeptides induced by type 1 and type 2 strains of herpes simplex virus. Intervirology. 1977;8(1):18–29. doi: 10.1159/000148873. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Purifoy D. J. Comparison of DNA polymerase activities induced by herpes simplex virus types 1 and 2. Intervirology. 1975;6(6):356–366. doi: 10.1159/000149492. [DOI] [PubMed] [Google Scholar]
  18. Purifoy D. J., Powell K. L. DNA-binding proteins induced by herpes simplex virus type 2 in HEp-2 cells. J Virol. 1976 Aug;19(2):717–731. doi: 10.1128/jvi.19.2.717-731.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schaffer P. A. Temperature-sensitive mutants of herpesviruses. Curr Top Microbiol Immunol. 1975;70:51–100. doi: 10.1007/978-3-642-66101-3_3. [DOI] [PubMed] [Google Scholar]
  20. Subak-Sharpe J. H., Brown S. M., Ritchie D. A., Timbury M. C., Macnab J. C., Marsden H. S., Hay J. Genetic and biochemical studies with herpesvirus. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 2):717–730. doi: 10.1101/sqb.1974.039.01.085. [DOI] [PubMed] [Google Scholar]
  21. Summers W. P., Wagner M., Summers W. C. Possible peptide chain termination mutants in thymide kinase gene of a mammalian virus, herpes simplex virus. Proc Natl Acad Sci U S A. 1975 Oct;72(10):4081–4084. doi: 10.1073/pnas.72.10.4081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Watson D. H., Shedden W. I., Elliot A., Tetsuka T., Wildy P., Bourgaux-Ramoisy D., Gold E. Virus specific antigens in mammalian cells infected with herpes simplex virus. Immunology. 1966 Oct;11(4):399–408. [PMC free article] [PubMed] [Google Scholar]
  23. Weissbach A., Hong S. C., Aucker J., Muller R. Characterization of herpes simplex virus-induced deoxyribonucleic acid polymerase. J Biol Chem. 1973 Sep 25;248(18):6270–6277. [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES