1887

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Volume 73, Issue 6

Expression of the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10) is required for virus growth and neoplastic transformation Free

Abstract

The amino-terminal domain of the large subunit of herpes simplex virus type 2 (HSV-2) ribonucleotide reductase (ICP10) has protein kinase (PK) activity and properties similar to those of growth factor receptor kinases which can be activated to transforming potential. DNA sequences that encode the PK domain cause neoplastic transformation of immortalized cells. The studies described in this report used a spontaneous mutant (5-152) temperature-sensitive for the synthesis of ICP10 and the previously described ICP10 expression vectors to study the role of ICP10 expression in HSV-2 growth and neoplastic potential. The titres of the 5-152 mutant are 1000-fold lower at 39 °C compared to 34 °C after 12 h post-infection. The efficiency of plaquing is 0.003. The growth defect at 39 °C correlates with decreased ICP10 synthesis. Sequence analysis of the PK domain of the 5-152 ICP10 gene identified a pair of frameshift mutations resulting in a 19 amino acid residue substitution at positions 275 to 293 and a downstream single base pair mutation causing a substitution at position 309. Cloning of the mutant ICP10 gene from 5-152 into a wild-type HSV-2 isolate resulted in a recombinant (859/152) with growth properties and rates of ICP10 synthesis at 39 °C similar to those of 5-152. Cells transformed with u.v.-inactivated 5-152, or the recombinant 859/152, have significantly decreased cloning efficiency in agarose at 39 °C, but only during the first 250 post-transfer population doublings. Anchorage-independent growth was observed in cells transfected with expression vectors pJW17 or pJW32 that express ICP10 or its PK domain, respectively. Cells transfected with the frameshift mutant pJW21 or the ICP10 carboxy-terminal vector pJW31 did not form clones in agarose.

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© Journal of General Virology 1992
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1992-06-01
2024-04-25
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References

  1. Anderson K. P., Frink R. J., Devi G. B., Gaylord B. H., Wagner E. K. 1981; Detailed characterization of the mRNA mapping in the Hind III fragment K region of the herpes simplex virus type 1 genome. Journal of Virology 37:1011–1027
     [Google Scholar]
  2. Aurelian L. 1968; Demonstration by plaque reduction technique of immunologic relationship between canine herpes virus and herpes simplex virus. Proceedings of the Society for Experimental Biology and Medicine 127:485–488
     [Google Scholar]
  3. Aurelian L., Terzano P., Smith C. C., Chung T. D., Shamsuddin A., Costa S., Orlandi C. 1989; Amino-terminal epitope of herpes simplex virus type 2 ICP10 protein as a molecular diagnostic marker for cervical intraepithelial neoplasia. Cancer Cells 7:187–191
     [Google Scholar]
  4. Aurelian L., Wachsman M., Burnett J. W. 1990; Clinical and subclinical HSV infection resulting from exposure to asymptomatic patients. British Journal of Dermatology 122:117–119
     [Google Scholar]
  5. Bacchetti S., Evelegh M. J., Muirhead B., Sartari C. S., Huszar D. 1984; Immunological characterization of herpes simplex virus type 1 and 2 polypeptide(s) involved in viral ribonucleotide reductase activity. Journal of Virology 49:591–593
     [Google Scholar]
  6. Barrett J. C., Crawford B. D., Mixter L. O., Schechtman L. M., Ts’o P. O. P., Pollack R. 1979; Correlation of in vitro growth properties and tumorigenicity of Syrian hamster cell lines. Cancer Research 39:1504–1510
     [Google Scholar]
  7. Chung T. D., Wymer J. P., Smith C. C., Kulka M., Aurelian L. 1989; Protein kinase activity associated with the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10). Journal of Virology 63:3389–3398
     [Google Scholar]
  8. Chung T. D., Wymer J. P., Kulka M., Smith C. C., Aurelian L. 1990; Myristylation and polylysine-mediated activation of the protein kinase domain of the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10). Virology 179:168–178
     [Google Scholar]
  9. Chung T. D., Luo J., Wymer J. P., Smith C. C., Aurelian L. 1991; Leucine repeats in the large subunit of herpes simplex virus type 2 ribonucleotide reductase (RR; ICP10) are involved in RR activity and subunit complex formation. Journal of General Virology 72:1139–1144
     [Google Scholar]
  10. Frame M. C., Marsden H. S., Dutia B. M. 1985; The ribonucleotide reductase induced by herpes simplex virus type 1 involves minimally a complex of two polypeptides (136K and 38K). Journal of General Virology 66:1581–1587
     [Google Scholar]
  11. Garnier J., Osguthorpe D. J., Robson B. 1978; Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. Journal of Molecular Biology 120:97–120
     [Google Scholar]
  12. Gascuel O., Golmard J. L. 1988; A simple method for predicting the secondary structure of globular proteins. Implications and accuracy. Computer Applications in the Biosciences 4:357–365
     [Google Scholar]
  13. Goldstein D. J., Weller S. K. 1988a; The HSV-l-induced ribonucleotide reductase activity is dispensable for virus growth and DNA synthesis: isolation and characterization of an ICP6 lacZ insertion mutant. Journal of Virology 62:195–205
     [Google Scholar]
  14. Goldstein D. J., Weller S. K. 1988b; Factor(s) present in herpes simplex virus type 1-infected cells can compensate for the loss of the large subunit of the viral ribonucleotide reductase: characterization of an ICP6 deletion mutant. Virology 166:41–51
     [Google Scholar]
  15. Hatada E., Hasegawa M., Shimizu K., Hatanaka M., Fukuda R. 1990; Analysis of influenza A virus temperature-sensitive mutants with mutations in RNA segment 8. Journal of General Virology 71:1283–1292
     [Google Scholar]
  16. Hayashi Y., Iwasaka T., Smith C. C., Aurelian L., Lewis G. K., Ts’o P. O. P. 1985; Multistep transformation by defined fragments of herpes simplex virus type 2 DNA: oncogenic region and its gene products. Proceedings of the National Academy of Sciences, U.S.A. 82:8493–8497
     [Google Scholar]
  17. Huszar D., Bacchetti S. 1981; Partial purification and characterization of the ribonucleotide reductase induced by herpes simplex virus infection of mammalian cells. Journal of Virology 37:580–588
     [Google Scholar]
  18. Iwasaka T., Smith C. C., Aurelian L., Ts’o P. O. P. 1985; The cervical tumor associated antigen (ICP10/AG-4) is encoded by the transforming region of the genome of herpes simplex virus type 2. Japanese Journal of Cancer Research 76:946–958
     [Google Scholar]
  19. Jariwalla R. J., Aurelian L., Ts’o P. O. P. 1980; Tumorigenic transformation induced by a specific fragment of DNA from herpes simplex virus type 2. Proceedings of the National Academy of Sciences, U.S.A. 77:2279–2283
     [Google Scholar]
  20. Jariwalla R. J., Aurelian L., Ts’o P. O. P. 1983; Immortalization and neoplastic transformation of normal diploid cells by defined cloned DNA fragments of herpes simplex virus type 2. Proceedings of the National Academy of Sciences, U.S.A. 80:5902–5906
     [Google Scholar]
  21. Jones C. 1989; The minimal transforming fragment of HSV-2 mtrIII can function as a complex promoter element. Virology 169:346–353
     [Google Scholar]
  22. Kelly K., Cochran B. H., Stiles C. D., Leder P. 1983; Cell-specific regulation of the c-myc gene by lymphocytes, mitogens and platelet-derived growth factor. Cell 35:603–610
     [Google Scholar]
  23. Knife D. M., Ruyechan W. I., Roizman B. 1979; Molecular genetics of herpes simplex virus. III. Fine mapping of a genetic locus determining resistance to phosphonoacetate. Journal of Virology 29:698–704
     [Google Scholar]
  24. Luo J. H., Aurelian L. 1992; The transmembrane helical segment but not the invariant lysine is required for the kinase activity of the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10). Journal of Biological Chemistry 267: (in press)
    [Google Scholar]
  25. Luo J. H., Smith C. C., Kulka M., Aurelian L. 1991; A truncated protein kinase domain of the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10) expressed in Escherichia coli. Journal of Biological Chemistry 266:20976–20983
     [Google Scholar]
  26. McLauchlan J., Clements J. B. 1983; DNA sequence homology between two co-linear loci on the HSV genome which have different transforming abilities. EMBO Journal 2:1953–1961
     [Google Scholar]
  27. Macnab J. C. M. 1974; Transformation of rat embryo cells by temperature-sensitive mutants of herpes simplex virus. Journal of General Virology 24:143–153
     [Google Scholar]
  28. Manak M., Aurelian L., Ts’o P. O. P. 1981; Focus formation and neoplastic transformation by herpes simplex virus type 2 inactivated intracellularly by BUdR and near UV light. Journal of Virology 40:289–300
     [Google Scholar]
  29. Matthews B. W., Nicholson H., Becktel W. J. 1987; Enhanced protein thermostability from site-directed mutations that decrease the entropy of unfolding. Proceedings of the National Academy of Sciences U.S.A. 84:6663–6667
     [Google Scholar]
  30. Novotny J., Auffray C. 1984; A program for prediction of protein secondary structure from nucleotide sequence data: application to histocompatibility antigens. Nucleic Acids Research 12:243–255
     [Google Scholar]
  31. Pignatti P. F., Cassai E., Meneguzzi G., Chenciner N., Milanesi G. 1979; Herpes simplex virus DNA isolation from infected cells with a novel procedure. Virology 93:260–264
     [Google Scholar]
  32. 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]
  33. Preston V. G., Darling A. A., McDougall I. M. 1988; The herpes simplex virus type 1 temperature sensitive mutant tsl222 has a single base pair deletion in the small subunit of ribonucleotide reductase. Virology 167:458–467
     [Google Scholar]
  34. Rapp F., Duff R. 1973; Transformation of hamster embryo fibroblasts by herpes simplex virus type 1 and 2. Cancer Research 33:1527–1534
     [Google Scholar]
  35. Roizman B., Norrild B., Chan C., Pereira L. 1984; Identification and preliminary mapping with monoclonal antibodies of a herpes simplex virus 2 glycoprotein lacking a known type 1 counterpart. Virology 133:242–247
     [Google Scholar]
  36. Saito T., Yamanaka K., Watanabe Y., Takamatsu N., Meshi T., Okada Y. 1989; Mutational analysis of the coat protein gene of tobacco mosaic virus in relation to hypersensitive response in tobacco plants with the N′ gene. Virology 173:11–20
     [Google Scholar]
  37. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, U.S.A. 74:5463–5467
     [Google Scholar]
  38. Smith C. C., Wymer J. P., Luo J. H., Aurelian L. 1991; Genomic sequences homologous to the protein kinase region of the bifunctional herpes simplex virus type 2 protein ICP10. Virus Genes 53:215–226
     [Google Scholar]
  39. 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]
  40. Swain M., Galloway D. A. 1986; Herpes simplex virus specifies two subunits of ribonucleotide reductase encoded by 3′-coterminal transcripts. Journal of Virology 57:802–808
     [Google Scholar]
  41. Wymer J. P., Chung T. D., Chang Y. N., Hayward G. S., Aurelian L. 1989; Identification of immediate-early-type cis-response elements in the promoter for the ribonucleotide reductase large subunit from herpes simplex virus type 2. Journal of Virology 63:2773–2784
     [Google Scholar]
  42. Wymer J. P., Aprhys C. M. J., Chung T. D., Feng C. P., Kulka M., Aurelian L. 1992; Immediate early and functional AP-1 cis-response elements are involved in the transcriptional regulation of the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10). Virus Research (in press)
    [Google Scholar]
  43. Yarden Y., Ullrich A. 1988; Growth factor receptor tyrosine kinases. Annual Review of Biochemistry 57:443–478
     [Google Scholar]
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Expression of the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10) is required for virus growth and neoplastic transformation
J. Gen. Virol. 73, 1417 (1992); https://doi.org/10.1099/0022-1317-73-6-1417
/content/journal/jgv/10.1099/0022-1317-73-6-1417
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