1887

Abstract

Summary

Herpes simplex virus (HSV) type 2 (strain HG52) has four I sites at map coordinates (m.c.) 0.45, 0.7, 0.91 and 0.94, i.e. two in the unique long and two in the unique short regions of the genome. Previously, we had isolated a genome containing only the 0.45 and 0.94 I sites. Here we report the isolation of a mutant (JH2611) in which all four I sites have been removed using an enrichment selection procedure, without any loss of viability. Removal of each site has been shown to be due to a base change or small undetectable deletion/insertion of less than 100 bp. In HSV-1, the I site at 0.45 m.c. is in an open reading frame coding for a polypeptide of 14K. Both the 0.7 and 0.94 m.c. HSV-2 I sites are in intergenic positions. The 0.91 m.c. I site has been shown to be within the coding sequence of the glycoprotein gG-2. Synthesis of gG-2 by JH2611 and two other mutants, JH2610 (formerly HG52X163X3) and JH2609 (formerly HG52X163X21), in which the 0.91 m.c. site has been deleted was analysed by immunoprecipitation using the gG-2-specific monoclonal antibodies AP1 and LP5 and the anti-peptide serum 14713. In the mutants JH2610 and JH2611 neither gG-2 nor its precursor were detected but the monoclonal antibodies detected two polypeptides migrating above the normal positions of gG-2 and the gG-2 precursor; these were not precipitated by the anti-peptide serum. With the mutant JH2609 neither gG-2 nor its precursors could be detected by either the monoclonal antibodies or the anti-peptide serum. The results strongly suggest that gG-2 is non-essential for the growth of HSV-2 .

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-69-1-113
1988-01-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/69/1/JV0690010113.html?itemId=/content/journal/jgv/10.1099/0022-1317-69-1-113&mimeType=html&fmt=ahah

References

  1. BALACHANDRAN N., HUTT-FLETCHER L. H. 1985; Synthesis and processing of glycoprotein gG of herpes simplex virus type 2. Journal of Virology 54:825–832
    [Google Scholar]
  2. BROWN S. M., HARLAND J. 1987; Three mutants of herpes simplex virus type 2: one lacking the genes US10, US11 and US12 and two in which Rs has been extended by 6 kb to 0·91 map units with loss of Us sequences between 0·94 and the US/TRS junction. Journal of General Virology 68:1–18
    [Google Scholar]
  3. BROWN S. M., RITCHIE D. A. 1975; Genetic studies of herpes simplex virus type 1. Analysis of mixed plaque-forming virus and its bearing on genetic recombination. Virology 64:32–42
    [Google Scholar]
  4. BROWN S. M., RITCHIE D. A., SUBAK-SHARPE J. H. 1973; Genetic studies with herpes simplex virus type 1. 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]
  5. BROWN S. M., HARLAND J., SUBAK-SHARPE J. H. 1984; Isolation of restriction endonuclease site deletion mutants of herpes simplex virus. Journal of General Virology 65:1053–1068
    [Google Scholar]
  6. COOK S. D., BROWN S. M. 1987; Herpes simplex virus type 1 latency in rabbit corneal cells in vitro: reactivation and recombination following intratypic superinfection of long term cultures. Journal of General Virology 68:813–824
    [Google Scholar]
  7. CORTINI R., WILKIE N. M. 1978; Physical maps for HSV type 2 DNA with five restriction endonucleases. Journal of General Virology 39:259–280
    [Google Scholar]
  8. DAVISON A. J., McGEOCH D. J. 1986; Evolutionary comparisons of the S segments in the genomes of herpes simplex virus type 1 and varicella-zoster virus. Journal of General Virology 67:597–611
    [Google Scholar]
  9. FRAME M. C, MARSDEN H. S., McGEOCH D. J. 1986; Novel herpes simplex virus type 1 glycoproteins identified by antiserum against a synthetic oligopeptide from the predicted product of gene US4. Journal of General Virology 67:745–751
    [Google Scholar]
  10. HARLAND J., BROWN S. M. 1985; Isolation and characterization of deletion mutants of herpes simplex virus type 2 (strain HG52). Journal of General Virology 66:1305–1321
    [Google Scholar]
  11. JONES W., SHENK T. 1978; Isolation of deletion and substitution mutants of adenovirus 5. Cell 13:181–188
    [Google Scholar]
  12. LONGNECKER R., ROIZMAN B. 1987; Clustering of genes dispensable for growth in culture in the S component of the HSV-1 genome. Science 236:573–576
    [Google Scholar]
  13. LONSDALE D. M. 1979; A rapid technique for distinguishing herpes simplex virus type 1 from type 2 by restriction enzyme technology. Lancet i:849–852
    [Google Scholar]
  14. McGEOCH D. J. 1984 The nature of animal virus genetic material. The Microbe 1984. Part I: Viruses75–107 Edited by Mahy B. W. J., Pattison J. R. Cambridge: Cambridge University Press;
    [Google Scholar]
  15. McGEOCH D. J., MOSS H. W. M., McNAB D., FRAME M. C. 1987; DNA sequence and genetic content of the HindIIIl region in the short unique component of the herpes simplex virus type 2 genome: identification of the gene encoding glycoprotein G, and evolutionary comparisons. Journal of General Virology 68:19–38
    [Google Scholar]
  16. MacLean A. R., BROWN S. M. 1987a; Generation of a herpes simplex virus type 1 variant devoid of XbaI sites. Journal of General Virology 68:1165–1171
    [Google Scholar]
  17. MacLean A. R., BROWN S. M. 1987b; A herpes simplex virus type 1 variant which fails to synthesize the immediate early polypeptide VmwIE63. Journal of General Virology 68:1339–1350
    [Google Scholar]
  18. MacLean A. R., BROWN S. M. 1987c; Deletion and duplication variants around the long repeats of herpes simplex virus type 1 strain 17. Journal of General Virology 68:3019–3031
    [Google Scholar]
  19. MARSDEN H. S., STOW N. D., PRESTON V. G., TIMBURY M. C., WILKIE N. M. 1978; Physical mapping of herpes simplex virus induced polypeptides. Journal of Virology 28:628–642
    [Google Scholar]
  20. MARSDEN H. S., BUCKMASTER A., PALFREYMAN J. W., HOPE R. G., MINSON A. C. 1984; Characterisation of the 92,000 dalton glycoprotein induced by herpes simplex virus type 2. Journal of Virology 50:547–551
    [Google Scholar]
  21. OLOFSSON S., LUNDSTRÖM M., MARSDEN H., JEANSSON S., VAHLNE A. 1986; Characterization of a herpex simplex virus type 2-specified glycoprotein with affinity for N-acetylgalactosamine-specific lectins and its identification as g92K or gG. Journal of General Virology 67:737–744
    [Google Scholar]
  22. ROIZMAN B., NORRILD B., CHAH C., PEREIRA L. 1984; Identification and preliminary mapping with monoclonal antibodies of a herpes simplex virus type 2 glycoprotein lacking a known type 1 counterpart. Virology 133:242–247
    [Google Scholar]
  23. SCHAFFER P. A., TELETHIA M. J., BENYESH-MELNICK M. 1974; Recombination between temperature-sensitive mutants of herpes simplex virus type 1. Virology 58:219–228
    [Google Scholar]
  24. STOW N. D. 1981; Cloning of a DNA fragment from the left hand terminus of the adenovirus type 5 genome and its use in site-directed mutagenesis. Journal of Virology 37:171–180
    [Google Scholar]
  25. STOW N. D., WILKIE N. M. 1976; An improved technique for obtaining enhanced infectivity with herpes simplex virus type 1 DNA. Journal of General Virology 33:447–458
    [Google Scholar]
  26. TIMBURY M. C. 1971; Temperature-sensitive mutants of herpes simplex virus type 2. Journal of General Virology 13:373–376
    [Google Scholar]
  27. UMENE K. 1985; Intermolecular recombination of the herpes simplex type 1 genome analysed using two strains differing in restriction enzyme cleavage sites. Journal of General Virology 66:2659–2670
    [Google Scholar]
  28. WHITTON J. L. 1984 Structural elements involved in herpes simplex virus immediate-early transcription Ph.D. thesis, University of Glasgow
    [Google Scholar]
  29. WILKIE N. M. 1973; The synthesis and substructure of herpesvirus DNA: the distribution of alkali-labile single strand interruptions in HSV-1 DNA. Journal of General Virology 21:453–467
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-69-1-113
Loading
/content/journal/jgv/10.1099/0022-1317-69-1-113
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