Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1983 Jan;45(1):343–353. doi: 10.1128/jvi.45.1.343-353.1983

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.

R A Dixon, D J Sabourin, P A Schaffer
PMCID: PMC256416  PMID: 6296441

Abstract

Eleven temperature-sensitive mutants of herpes simplex virus type 2 (HSV-2) exhibit overlapping patterns of complementation that define four functional groups. Recombination tests confirmed the assignment of mutants to complementation groups 1 through 4 and permitted the four groups to be ordered in an unambiguous linear array. Combined recombination and marker rescue tests (A. E. Spang, P. J. Godowski, and D. M. Knipe, J. Virol. 45:332-342, 1983) indicate that the mutations lie in a tight cluster near the center of UL to the left of the gene for DNA polymerase in the order 4-3-2-1-polymerase. The seven mutants that make up groups 1 and 2 fail to complement each other and mutants in HSV-1 complementation group 1-1, the group thought to define the structural gene for the major HSV-1 DNA-binding protein with a molecular weight of 130,000. At 38 degrees C, mutants in groups 1 and 2 synthesize little or no viral DNA, and unlike cells infected with the wild-type virus, mutant-infected cells exhibit no detectable nuclear antigen reactive with monoclonal or polypeptide-specific antibody to the major HSV-2 DNA-binding protein. The four mutants that make up groups 3 and 4 do not complement each other, nor do they complement mutants in group 2. They do, however, complement mutants in group 1 as well as representative mutants of HSV-1 complementation group 1-1. At 38 degrees C, mutants in groups 3 and 4 are phenotypically DNA+, and nuclei of mutant-infected cells contain the HSV-2 DNA-binding protein. Thus, the four functional groups appear to define two closely linked genes, one encoding an early viral function affecting both viral DNA synthesis and expression of the DNA-binding protein with a molecular weight of 130,000 (groups 1 and 2), and the other encoding a previously unidentified late viral function (groups 3 and 4). The former gene presumably represents the structural gene for the major HSV-2 DNA-binding protein.

Full text

PDF
343

Images in this article

349Image
on p.349

Selected References

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

  1. Aron G. M., Purifoy D. J., Schaffer P. A. DNA synthesis and DNA polymerase activity of herpes simplex virus type 1 temperature-sensitive mutants. J Virol. 1975 Sep;16(3):498–507. doi: 10.1128/jvi.16.3.498-507.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chartrand P., Crumpacker C. S., Schaffer P. A., Wilkie N. M. Physical and genetic analysis of the herpes simplex virus DNA polymerase locus. Virology. 1980 Jun;103(2):311–326. doi: 10.1016/0042-6822(80)90190-7. [DOI] [PubMed] [Google Scholar]
  3. Chartrand P., Stow N. D., Timbury M. C., Wilkie N. M. Physical mapping of paar mutations of herpes simplex virus type 1 and type 2 by intertypic marker rescue. J Virol. 1979 Aug;31(2):265–276. doi: 10.1128/jvi.31.2.265-276.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chartrand P., Timbury M. C., Hay J., Moss H. Mutant of herpes simplex virus type 2 with temperature-sensitive lesions affecting virion thermostability and DNase activity: identification of the lethal mutation and physical mapping of the nuc-lesion. J Virol. 1979 Oct;32(1):140–146. doi: 10.1128/jvi.32.1.140-146.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chu C. T., Schaffer P. A. Qualitative complementation test for temperature-sensitive mutants of herpes simplex virus. J Virol. 1975 Nov;16(5):1131–1136. doi: 10.1128/jvi.16.5.1131-1136.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Conley A. J., Knipe D. M., Jones P. C., Roizman B. Molecular genetics of herpes simplex virus. VII. Characterization of a temperature-sensitive mutant produced by in vitro mutagenesis and defective in DNA synthesis and accumulation of gamma polypeptides. J Virol. 1981 Jan;37(1):191–206. doi: 10.1128/jvi.37.1.191-206.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Courtney R. J., Benyesh-Melnick M. Isolation and characterization of a large molecular-weight polypeptide of herpes simplex virus type 1. Virology. 1974 Dec;62(2):539–551. doi: 10.1016/0042-6822(74)90414-0. [DOI] [PubMed] [Google Scholar]
  8. Esparza J., Benyesh-Melnick B., Schaffer P. A. Intertypic complementation and recombination between temperature-sensitive mutants of herpes simplex virus types 1 and 2. Virology. 1976 Apr;70(2):372–384. doi: 10.1016/0042-6822(76)90279-8. [DOI] [PubMed] [Google Scholar]
  9. Esparza J., Purifoy D. J., Schaffer P. A., Benyesh-Melnick M. Isolation, complementation and preliminary phenotypic characterization of temperature-sensitive mutants of herpes simplex virus type 2. Virology. 1974 Feb;57(2):554–565. doi: 10.1016/0042-6822(74)90194-9. [DOI] [PubMed] [Google Scholar]
  10. Flannery V. L., Courtney R. J., Schaffer P. A. Expression of an early, nonstructural antigen of herpes simplex virus in cell transformed in vitro by herpes simplex virus. J Virol. 1977 Jan;21(1):284–291. doi: 10.1128/jvi.21.1.284-291.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Frenkel N., Locker H., Vlazny D. A. Studies of defective herpes simplex viruses. Ann N Y Acad Sci. 1980;354:347–370. doi: 10.1111/j.1749-6632.1980.tb27977.x. [DOI] [PubMed] [Google Scholar]
  12. Halliburton I. W., Morse L. S., Roizman B., Quinn K. E. Mapping of the thymidine kinase genes of type 1 and type 2 herpes simplex viruses using intertypic recombinants. J Gen Virol. 1980 Aug;49(2):235–253. doi: 10.1099/0022-1317-49-2-235. [DOI] [PubMed] [Google Scholar]
  13. Halliburton I. W., Randall R. E., Killington R. A., Watson D. H. Some properties of recombinants between type 1 and type 2 herpes simplex viruses. J Gen Virol. 1977 Sep;36(3):471–484. doi: 10.1099/0022-1317-36-3-471. [DOI] [PubMed] [Google Scholar]
  14. Kaerner H. C., Maichle I. B., Ott A., Schröder C. H. Origin of two different classes of defective HSV-1 Angelotti DNA. Nucleic Acids Res. 1979 Apr;6(4):1467–1478. doi: 10.1093/nar/6.4.1467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Little S. P., Jofre J. T., Courtney R. J., Schaffer P. A. A virion-associated glycoprotein essential for infectivity of herpes simplex virus type 1. Virology. 1981 Nov;115(1):149–160. doi: 10.1016/0042-6822(81)90097-0. [DOI] [PubMed] [Google Scholar]
  16. Marsden H. S., Stow N. D., Preston V. G., Timbury M. C., Wilkie N. M. Physical mapping of herpes simplex virus-induced polypeptides. J Virol. 1978 Nov;28(2):624–642. doi: 10.1128/jvi.28.2.624-642.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Morse L. S., Buchman T. G., Roizman B., Schaffer P. A. Anatomy of herpes simplex virus DNA. IX. Apparent exclusion of some parental DNA arrangements in the generation of intertypic (HSV-1 X HSV-2) recombinants. J Virol. 1977 Oct;24(1):231–248. doi: 10.1128/jvi.24.1.231-248.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Morse L. S., Pereira L., Roizman B., Schaffer P. A. Anatomy of herpes simplex virus (HSV) DNA. X. Mapping of viral genes by analysis of polypeptides and functions specified by HSV-1 X HSV-2 recombinants. J Virol. 1978 May;26(2):389–410. doi: 10.1128/jvi.26.2.389-410.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Porter D. D., Wimberly I., Benyesh-Melnick M. Prevalence of antibodies to EB virus and other herpesviruses. JAMA. 1969 Jun 2;208(9):1675–1679. [PubMed] [Google Scholar]
  20. Powell K. L., Littler E., Purifoy D. J. Nonstructural proteins of herpes simplex virus. II. Major virus-specific DNa-binding protein. J Virol. 1981 Sep;39(3):894–902. doi: 10.1128/jvi.39.3.894-902.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Preston C. M., Cordingley M. G. mRNA- and DNA-directed synthesis of herpes simplex virus-coded exonuclease in Xenopus laevis oocytes. J Virol. 1982 Aug;43(2):386–394. doi: 10.1128/jvi.43.2.386-394.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Reyes G. R., LaFemina R., Hayward S. D., Hayward G. S. Morphological transformation by DNA fragments of human herpesviruses: evidence for two distinct transforming regions in herpes simplex virus types 1 and 2 and lack of correlation with biochemical transfer of the thymidine kinase gene. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 1):629–641. doi: 10.1101/sqb.1980.044.01.066. [DOI] [PubMed] [Google Scholar]
  24. Schaffer P. A., Aron G. M., Biswal N., Benyesh-Melnick M. Temperature-sensitive mutants of herpes simplex virus type 1: isolation, complementation and partial characterization. Virology. 1973 Mar;52(1):57–71. doi: 10.1016/0042-6822(73)90398-x. [DOI] [PubMed] [Google Scholar]
  25. Schaffer P. A., Carter V. C., Timbury M. C. Collaborative complementation study of temperature-sensitive mutants of herpes simplex virus types 1 and 2. J Virol. 1978 Sep;27(3):490–504. doi: 10.1128/jvi.27.3.490-504.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schaffer P. A., Tevethia M. J., Benyesh-Melnick M. Recombination between temperature-sensitive mutants of herpes simplex virus type 1. Virology. 1974 Mar;58(1):219–228. doi: 10.1016/0042-6822(74)90156-1. [DOI] [PubMed] [Google Scholar]
  27. Spang A. E., Godowski P. J., Knipe D. M. Characterization of herpes simplex virus 2 temperature-sensitive mutants whose lesions map in or near the coding sequences for the major DNA-binding protein. J Virol. 1983 Jan;45(1):332–342. doi: 10.1128/jvi.45.1.332-342.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Takahashi M., Yamanishi K. Transformation of hamster embryo and human embryo cells by temperature sensitive mutants of herpes simplex virus type 2. Virology. 1974 Sep;61(1):306–311. doi: 10.1016/0042-6822(74)90267-0. [DOI] [PubMed] [Google Scholar]
  29. Timbury M. C., Subak-Sharpe J. H. Genetic interactions between temperature-sensitive mutants of types 1 and 2 herpes simplex viruses. J Gen Virol. 1973 Mar;18(3):347–357. doi: 10.1099/0022-1317-18-3-347. [DOI] [PubMed] [Google Scholar]
  30. Weller S. K., Lee K. J., Sabourin D. J., Schaffer P. A. Genetic analysis of temperature-sensitive mutants which define the gene for the major herpes simplex virus type 1 DNA-binding protein. J Virol. 1983 Jan;45(1):354–366. doi: 10.1128/jvi.45.1.354-366.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Zygraich N., Huygelen C. In vivo behaviour of a temperature-sensitive (ts) mutant of herpesvirus hominis type 2. Arch Gesamte Virusforsch. 1973;43(1):103–111. doi: 10.1007/BF01249353. [DOI] [PubMed] [Google Scholar]

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

RESOURCES