1887

Abstract

SUMMARY

Herpesvirus saimiri-specific proteins from the nuclear fractions of productively infected owl monkey kidney cells were dissociated from virus and host DNA by treatment with 2 -NaCl or separation on Urografin density gradients. Empty virus capsids remained intact and could be separated from major non-structural proteins (110K, 51K and 48K) and from a subset of structural proteins (130K, 29K and 12K), either by Urografin gradient sedimentation or differential centrifugation. The DNA in such soluble extracts of nuclear proteins was efficiently removed by spermine precipitation, together with the host cell histones and large fractions of the 130K and 12K structural proteins. Proteins in the spermine-soluble fraction were analysed by affinity chromatography on columns of single-stranded calf thymus DNA coupled to cellulose. Two major structural proteins (130K and 12K), whose synthesis was sensitive to phosphonoacetic acid (PAA), and one minor PAA-resistant structural protein (29K) bound to DNA-cellulose. The major PAA-resistant 110K non-structural protein and the PAA-resistant non-structural 51K and 48K phosphoproteins were efficiently released into the spermine-soluble fraction and also bound to DNA-cellulose as did the 76K protein and minor species of 42K, 39K, 34K, 25K and 21K. Virus-specific proteins were eluted from such columns by buffers containing 0.4 -NaCl or by heparin in lowsalt buffers. Polypeptides from virus particles, infected cell extracts, or samples of eluates from DNA-cellulose chromatography, were separated by SDS-polyacrylamide gel electrophoresis, transferred onto nitrocellulose filters and probed for their ability to bind labelled polynucleotides. The non-structural 51K phosphoprotein, the 12K and 29K structural proteins and a 100K virion polypeptide all bound labelled DNA. However, the binding activities of the 130K protein from virions or purified by affinity chromatography and of the 110K polypeptide could not be demonstrated reproducibly after transfer from SDS gels to nitrocellulose. Comparisons of the present results on the properties of the herpesvirus saimiri-specified DNA-binding proteins with published accounts of the DNA-binding proteins of other herpesviruses, suggest some striking similarities with the DNA-binding proteins of the Epstein—Barr virus.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-64-12-2697
1983-12-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/64/12/JV0640122697.html?itemId=/content/journal/jgv/10.1099/0022-1317-64-12-2697&mimeType=html&fmt=ahah

References

  1. Bayliss G. C., Marsden H. S., Hay J. 1975; Herpes simplex virus proteins: DNA-binding proteins in infected cells and in the virus structure. Virology 68:124–134
    [Google Scholar]
  2. Bowen B., Steinberg J., Laemmli U. K., Weintraub H. 1980; The detection of DNA-binding proteins by protein blotting. Nucleic Acids Research 8:1–20
    [Google Scholar]
  3. Desrosiers R. C. A., Falk L. A. JR 1981; Herpesvirus tamarinus and its relation to herpes simplex virus. Journal of General Virology 56:119–130
    [Google Scholar]
  4. 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]
  5. Falk L. A., Wolfe L. G., Deinhardt F. 1972; Isolation of herpesvirus saimiri from blood of squirrel monkeys (Saimiri sciureus). Journal of the National Cancer Institute 48:1499–1505
    [Google Scholar]
  6. Feighny R. J., Farrell M. P., Pagano J. S. 1980; Polypeptide synthesis and phosphorylation in Epstein-Barr virus-infected cells. Journal of Virology 34:455–463
    [Google Scholar]
  7. Frenkel N., Jacob R. J., Honess R. W., Hayward G. S., Locker H., Roizman B. 1975; Anatomy of herpes simplex virus DNA. III. Characterization of defective DNA molecules and biological properties of virus populations containing them. Journal of Virology 16:153–167
    [Google Scholar]
  8. Gibson W., Roizman B. 1972; Proteins specified by herpes simplex virus. VIII. Characterization and composition of multiple capsid forms of subtypes 1 and 2. Journal of Virology 10:1044–1052
    [Google Scholar]
  9. Gibson W., Roizman B. 1974; Proteins specified by herpes simplex virus. X. Staining and radiolabeling properties of B-capsids and virion polypeptides in polyacrylamide gels. Journal of Virology 13:155–165
    [Google Scholar]
  10. Heine J. W., Honess R. W., Cassai E., Roizman B. 1974; Proteins specified by herpes simplex virus. XII. The virion polypeptides of type 1 strains. Journal of Virology 14:640–651
    [Google Scholar]
  11. Honess R. W., Watson D. H. 1977; Unity and diversity in the herpesviruses. Journal of General Virology 37:15–37
    [Google Scholar]
  12. Honess R. W., O’Hare P., Young D. 1982; Comparison of thymidine kinase activities induced in cells productively infected with herpesvirus saimiri and herpes simplex virus. Journal of General Virology 58:237–249
    [Google Scholar]
  13. Hoopes B. C., Mcclure W. R. 1981; Studies on the selectivity of DNA precipitation by spermine. Nucleic Acids Research 9:5493–5504
    [Google Scholar]
  14. Kawanishi M., Sugawara K., Ito Y. 1981a; Epstein-Barr virus-induced polypeptides: a comparative study with superinfected Raji, lUdR-treated, and n-butyrate treated P3HR-1 cells. Virology 109:72–81
    [Google Scholar]
  15. Kawanishi M., Sugawara K., Ito Y. 1981b; Epstein-Barr virus-induced early polypeptides in Raji and NC37 cells activated by diterpene ester TPA in combination with n-butyrate. Virology 115:406–409
    [Google Scholar]
  16. Keil G., Muller I., Fleckenstein B., Koomey J. M., Mulder C. 1980; Generation of recombinants between different strains of herpesvirus saimiri. In Viruses in Naturally Occurring Cancers pp 145–161 Edited by Essex M., Todaro G., Zur Hausen H. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  17. Kelly R. B., Cozzarelli N., Deutscher M. P., Lehman I. P., Kornberg A. 1970; Enzymatic synthesis of deoxyribonucleic acid,. XXXII,. Replication of duplex deoxyribonucleic acid by polymerase at a single strand break. Journal of Biological Chemistry 245:39–45
    [Google Scholar]
  18. Lafer E. M., Moller A., Nordheim A., Stollar B. D., Rich A. 1981; Antibodies specific for left-handed Z-DNA,. Proceedings of the National Academy of Sciences, U. S. A 78:3546–3550
    [Google Scholar]
  19. Lemaster S., Roizman B. 1980; Herpes simplex virus phosphoproteins,. II,. Characterization of the virion protein kinase and of the polypeptides phosphorylated in the virion. Journal of Virology 35:798–811
    [Google Scholar]
  20. Litman R. M. 1968; A deoxyribonucleic acid polymerase from Micrococcus luteus (Micrococcus lysodeikticus) isolated on deoxyribonucleic acid-cellulose. Journal of Biological Chemistry 243:6222–6233
    [Google Scholar]
  21. Littler E., Purifoy D., 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]
  22. Modrow S., Wolf H. 1983; Herpesvirus saimiri-induced proteins in lytically infected cells. I. Time-ordered synthesis. Journal of General Virology 64:37–46
    [Google Scholar]
  23. Mueller-Lantzsch N., Yamamoto N. 1981; Epstein-Barr virus-induced proteins. IV. Characterization of an EBV-associated phosphopolypeptide. Journal of General Virology 55:333–341
    [Google Scholar]
  24. Mueller-Lantzsch N., Yamamoto N., Zur Hausen H. 1979; Analysis of early and late Epstein-Barr virus associated polypeptides by immunoprecipitation. Virology 97:378–387
    [Google Scholar]
  25. O’Hare P., Honess R. W. 1983a; Identification of a subset of herpesvirus saimiri polypeptides synthesized in the absence of virus DNA replication. Journal of Virology 46:279–283
    [Google Scholar]
  26. O’Hare P., Honess R. W. 1983b; 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]
  27. Powell K. L., Purifoy D. J. M. 1976; DNA-binding proteins of cells infected by herpes simplex virus type 1 and type 2. Intervirology 7:225–239
    [Google Scholar]
  28. 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]
  29. Powell K. L., Watson D. H. 1975; Some structural antigens of herpes simplex virus type 1. Journal of General Virology 29:167–178
    [Google Scholar]
  30. Powell K. L., Littler E., Purifoy D. J. M. 1981; Nonstructural proteins of herpes simplex virus,. II,. Major virus-specific DNA-binding protein. Journal of Virology 39:894–902
    [Google Scholar]
  31. Preston V. G., Coates J., Rixon F. J. 1983; Identification and characterization OF a herpes simplex virus gene product required for encapsidation of virus DNA. Journal of Virology 45:1056–1064
    [Google Scholar]
  32. Purifoy D. J. M., Powell K. L. 1976; DNA-binding proteins induced by herpes simplex virus type 2 in HEp-2 cells. Journal of Virology 19:717–731
    [Google Scholar]
  33. Randall R. E., Honess R. W. 1980; Proteins of herpesvirus saimiri: identification of two polypeptides released into the culture medium of productively infected cells. Journal of General Virology 51:445–449
    [Google Scholar]
  34. Randall R. E., Honess R. W., O’Hare P. 1983; Proteins specified by herpesvirus saimiri: identification and properties of virus-specified polypeptides in productively infected cells. Journal of Gerneral Virology 64:19–35
    [Google Scholar]
  35. Rigby P. W. J., Dieckmann M., Rhodes C., Berg P. 1977; Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. Journal of Molecular Biology 113:237–251
    [Google Scholar]
  36. Roubal J., Kallin B., Luka J., Klein G. 1981; Early DNA-binding polypeptides of Epstein-Barr virus. Virology 113:285–292
    [Google Scholar]
  37. Russell W. C., Precious B. 1982; Nucleic acid-binding properties of adenovirus structural polypeptides. Journal of General Virology 63:69–79
    [Google Scholar]
  38. Russell W. C., Mcintosh K., Skehel J. J. 1971; The preparation and properties of adenovirus cores. Journal of General Virology 11:35–46
    [Google Scholar]
  39. Schaffer P. A., Falk L. A., Deinhardt F. 1975; Attenuation of herpesvirus saimiri for marmosets after successive in cell culture at 39 °C. Journal of the National Cancer Institute 55:1243–1246
    [Google Scholar]
  40. Seed B. 1982; Diazotizable arylamine cellulose papers for the coupling and hybridization of nucleic acids. Nucleic Acids Research 10:1799–1810
    [Google Scholar]
  41. Spang A. E., Godowski P. J., Knipe D. M. 1983; Characterization of herpes simplex virus 2 temperature-sensitive mutants whose lesions map in or near the coding sequences for the major DNA-binding protein. Journal of Virology 45:332–342
    [Google Scholar]
  42. Spear P. G., Roizman B. 1972; Proteins specified BY herpes simplex virus. V. Purification and structural proteins of the herpesvirion. Journal of Virology 9:143–159
    [Google Scholar]
  43. Subirana J. A., Vives J. L. 1981; The precipitation of DNA by spermine. Biopolymers 20:2281–2283
    [Google Scholar]
  44. Sugawara K., Kawanishi M., Ito Y. 1982; Epstein-Barr virus-related DNA-binding proteins induced by n-butyrate in P3HR-1 cells. Virology 116:354–358
    [Google Scholar]
  45. 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]
  46. 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]
  47. 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]
  48. Yeo J., Killington R., Watson D. H., Powell K. 1981; Studies on cross-reactive antigens in the herpesviruses. Virology 108:256–266
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-64-12-2697
Loading
/content/journal/jgv/10.1099/0022-1317-64-12-2697
Loading

Data & Media loading...

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