1887

Abstract

SUMMARY

The polypeptides induced in cells infected with a Glasgow isolate of HSV-1 (17 ) have been characterized by SDS polyacrylamide gel electrophoresis.

Study of the kinetics of synthesis in three cell lines has detected a total of 52 polypeptides, 33 of which can be identified in polypeptide profiles of purified virions. These include six low mol. wt. polypeptides that have not been previously reported. Several polypeptides were labelled with glucosamine in infected BHK cells.

The different polypeptide patterns obtained at permissive (31 °C) and non-permissive (38 °C) temperature in cells infected with 16 temperature-sensitive () mutants are reported. The effect of multiplicity of infection (m.o.i.) on the polypeptide profile has been examined for two of the DNA — ve mutants: below ten, the profile varied with the m.o.i. whereas above ten it was constant. All mutants were therefore examined at an m.o.i. of approx. 20. Mutants from the same complementation group showed very similar profiles.

A number of general conclusions concerning control of protein synthesis in HSV infected cells can be made: (1) As most of the 16 mutants affected the synthesis of several or many polypeptides it follows that a large proportion of the genome specifies controlling functions. (2) The high frequency with which some polypeptides were affected suggests they are at or near the terminus of biosynthetic pathways which are under multiple control. (3) Conversely, some polypeptides were affected with a low frequency suggesting that their synthesis is not dependent on the expression of many virus functions. (4) Several individual mutations lead to the synthesis of increased amounts of different large polypeptides. (5) Analysis of every band detectably affected by at least one mutation has disclosed nine classes of dependence relationship between polypeptide synthesis and the DNA phenotype of the mutants, illustrating that this relationship is complex and different for different polypeptides. (6) The inhibition of host protein synthesis by the virus may not be a simple single step process.

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1976-06-01
2023-02-01
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References

  1. ABODEELY R. A., PALMER E., LAWSON L. A., RANDALL C. C. 1971; The proteins of enveloped and de-enveloped equine abortion (herpes) virus and separated envelope. Virology 44:146–152
    [Google Scholar]
  2. BAYLISS G. J., 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]
  3. BERG D., CHAMBERLIN M. 1970; Physical studies on ribonucleic acid polymerase from Escherichia coli B. Biochemistry 9:5055–5064
    [Google Scholar]
  4. BONE D. R., COURTNEY R. J. 1974; A temperature-sensitive mutant of herpes simplex virus type 1 defective in the synthesis of the major capsid polypeptide. Journal of General Virology 24:17–27
    [Google Scholar]
  5. BOLLUM R. J. 1968; Filter paper disc techniques for assaying radioactive macromolecules. Methods in Enzymology 12:169–173
    [Google Scholar]
  6. BROWN S. M., RITCHIE D. A., SUBAK-SHARPE J. M. 1973; Genetic studies with herpes simplex virus type i. The isolation of temperature-sensitive mutants, their arrangement into complementation groups and recombination analysis leading to a linkage map. Journal of General Virology 18:329–346
    [Google Scholar]
  7. BÜRK R. R. 1967 Growth Regulating Substances from Animal Cells in Culture39 Edited by Defendi V., Stoker M. G. P. Philadelphia, Pa: Wistar Institute Press;
    [Google Scholar]
  8. COURTNEY R. J., BENYESH-MELNICK M. 1974; Isolation and characterization of a large molecular-weight polypeptide of herpes simplex virus type 1. Virology 62:539–551
    [Google Scholar]
  9. ESPARZA J., PURIFOY D. J. M., SCHAFFER P. A., BENYESH-MELNICK M. 1974; Isolation, complementation and preliminary phenotypic characterization of temperature-sensitive mutants of herpes simplex virus type 2. Virology 57:554–565
    [Google Scholar]
  10. FAIRBANKS G. JUN., LEVINTHAL C., REEDER R. H. 1965; Analyses of (14C)-labelled protein by disc electrophoresis. Biochemical and Biophysical Research Communications 20:393–3991965
    [Google Scholar]
  11. FALKE D., HEICKE B., BASSLER R. 1972; The effect of arabinofuranosyl-cytosine upon the synthesis of herpes virus hominis. Electron microscopic observations in relation to viral DNA synthesis. Archiv fur die gesamte Virusforschung 39:48–62
    [Google Scholar]
  12. 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]
  13. HONESS R. W., ROIZMAN B. 1973; Proteins specified by herpes simplex virus. XL Identification and relative molar rates of synthesis of structural and nonstructural herpesvirus polypeptides in the infected cell. Journal of Virology 12:1347–1365
    [Google Scholar]
  14. HONESS R. W., ROIZMAN B. 1974; Regulation of herpesvirus macromolecular synthesis. I. Cascade regulation of the synthesis of three groups of viral proteins. Journal of Virology 14:8–19
    [Google Scholar]
  15. HOWARD D. K., HAY J., MELVIN W. A., DURHAM J. P. 1974; Changes in DNA and RNA synthesis and associated enzyme activities after the stimulation of serum-depleted BHK 21/C13 cells by the addition of serum. Experimental Cell Research 86:31–42
    [Google Scholar]
  16. KEMP C. M., PERDUE M. L., ROGERS H. W., O’CALLAGHAN D. J., RANDALL C. C. 1974; Structural polypeptides of the hamster strain of equine herpes virus type 1: products associated with purification. Virology 61:361–375
    [Google Scholar]
  17. KLOTZ I. M., DARNELL D. W. 1969; Protein subunits: a table (second edition). Science, New York 166:126–128
    [Google Scholar]
  18. LAEMMLI U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  19. MACPHERSON I., STOKER M. 1962; Polyoma transformation of hamster cell clones. An investigation of genetic factors affecting cell competence. Virology 16:147–151
    [Google Scholar]
  20. MARGOLIS J., KENRICK K. G. 1967; Polyacrylamide gel-electrophoresis across a molecular sieve gradient. Nature, London 214:1334–1336
    [Google Scholar]
  21. MECHIE M. M. 1974; A biological and biochemical characterisation of ts mutants of herpes simplex virus type 1. Ph.D. Thesis, University of Glasgow
    [Google Scholar]
  22. NII S., ROZENKRANZ S., MORGAN C., ROSE H. M. 1968; Electron microscopy of herpes simplex virus. III. Effect of hydroxyurea. Journal of Virology 2:1163–1171
    [Google Scholar]
  23. OLSHEVESKY U., LEVITT J., BECKER R. 1967; Studies on the synthesis of herpes simplex virions. Virology 33:323–334
    [Google Scholar]
  24. OLSHEVESKY U., BECKER R. 1970; Herpes simplex virus structural proteins. Virology 40:948–960
    [Google Scholar]
  25. PRINGLE C. R., HOWARD D. K., HAY J. 1973; Temperature-sensitive mutants of Pseudorabies virus with differential effects on viral and host DNA synthesis. Virology 55:495–505
    [Google Scholar]
  26. RAKUSANOVA T., BEN-PORAT T., HIMENO M., KAPLAN A. S. 1971; Early function of the genome of herpesvirus. I. Characterization of the RNA synthesized in cycloheximide-treated, infected cells. Virology 46:877–889
    [Google Scholar]
  27. ROBINSON D. J., WATSON D. H. 1971; Structural proteins of herpes simplex virus. Journal of General Virology 10:163–171
    [Google Scholar]
  28. SCHAFFER P., VONKA V., LEWIS R., BENYESH-MELNICK M. 1970; Temperature-sensitive mutants of herpes simplex virus. Virology 42:1144–1146
    [Google Scholar]
  29. SCHAFFER P. A., COURTNEY R. J., MCCOMBS R. M., BENYESH-MELNICK M. 1971; A temperature-sensitive mutant of herpes simplex virus defective in glycoprotein synthesis. Virology 46:356–368
    [Google Scholar]
  30. SCHAFFER P. A., ARON G. M., BISWAL N., BENYESH-MELNICK M. 1973; Temperature-sensitive mutants of herpes simplex virus type 1: isolation, complementation and partial characterization. Virology 52:57–71
    [Google Scholar]
  31. SCHAFFER P.A., BRUNSCHWIG J. P., MCCOMBS R. M., BENYESH-MELNICK M. 1974; Electron microscopic studies of temperature-sensitive mutants of herpes simplex virus type 1. Virology 62:444–457
    [Google Scholar]
  32. SHAPIRO A. L., VINUELA E., MAIZEL J. V. 1967; Molecular weight estimation of polypeptide chains by electrophoresis in SDS Polyacrylamide gels. Biochemical and Biophysical Research Communications 28:815–820
    [Google Scholar]
  33. SHIMONO H., BEN-PORAT T., KAPLAN A. S. 1969; Synthesis of proteins in cells infected with herpesvirus. 1. Structural viral proteins. Virology 37:49–55
    [Google Scholar]
  34. SPEAR P. G., ROIZMAN B. 1972; Proteins specified by herpes simplex virus. V. Purification and structural proteins of the herpes virion. Journal of Virology 9:143–159
    [Google Scholar]
  35. SPEAR P. G. 1975; Glycoproteins specified by herpes simplex virus type 1: herpesvirion envelopes and infected cell membranes, their synthesis, processing and antigenic relatedness to herpes simplex type 2. Proceedings of the Conference on Herpesviruses and OncogenesisNuremberg1974 Edited by Zur Hausen H., Epstein A., de The G. International Agency for Research on Cancer49–61
    [Google Scholar]
  36. STUDIER F. W. 1973; Analysis of bacteriophage T7 early RNAs and proteins on slab gels. Journal of Molecular Biology 79:237–248
    [Google Scholar]
  37. SUBAK-SHARPE J. H. 1969 Proceedings of the First International Congress for Virology, Helsinki252 Edited by Melnick J. L. Basel: Karger;
    [Google Scholar]
  38. SUBAK-SHARPE J. H., BROWN S. M., RITCHIE D. A., TIMBURY M. C, MCNAB J. C. M., MARSDEN H. S., HAY J. 1974; Genetic and biochemical studies with herpesvirus. Cold Spring Harbor Symposium on Quantitative Biology 39:717–739
    [Google Scholar]
  39. TAKAHASHI M., YAMANISHI K. 1974; Transformation of hamster embryo and human embryo cells by temperature-sensitive mutants of herpes simplex virus type 2. Virology 61:306–311
    [Google Scholar]
  40. TIMBURY M. C. 1971; Temperature-sensitive mutants of herpes simplex virus type 2. Journal of General Virology 13:373–376
    [Google Scholar]
  41. TRAVERS A. A., BURGESS R. R. 1969; Cyclic re-use of the RNA polymerase sigma factor. Nature, London 222:537–540
    [Google Scholar]
  42. WARD L. R., STEVENS J. G. 1975; Effect of cytosine arabinoside on viral-specific protein synthesis in cells infected with herpes simplex virus. Journal of Virology 15:71–80
    [Google Scholar]
  43. WEBER K., OSBORN M. 1969; The reliability of molecular weight determinations by dodecyl sulfate -Polyacrylamide gel electrophoresis. Journal of Biological Chemistry 244:4406–4412
    [Google Scholar]
  44. WEEDS A. G., LOWEY S. 1971; Substructure of the myosin molecule. II. The light chains of myosin. Journal of Molecular Biology 61:701–725
    [Google Scholar]
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