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. 1981 Aug;39(2):559–572. doi: 10.1128/jvi.39.2.559-572.1981

Herpes simplex virus type 1 HindIII fragment L encodes spliced and complementary mRNA species.

R J Frink, K P Anderson, E K Wagner
PMCID: PMC171366  PMID: 6268843

Abstract

We have used DNA bound to cellulose to isolate and translate in vitro herpes simplex virus type 1 (HSV-1) mRNA's encoded by HindIII fragment L (mapping between 0.592 and 0.647), and 8.450-base-pair (8.45-kb) portion of the long unique region of the viral genome. Readily detectable, late mRNA's 2.7 and 1.9 kb in size encoding 69,000- and 58,000-dalton polypeptides, respectively, were isolated. A very minor late mRNA family composed of two colinear forms, one 2.6 kb and one 2.8 kb, was isolated and found to encode only an 85,000-dalton polypeptide. A major early mRNA, 1.8 kb in size encoding a 64,000-dalton polypeptide, was also isolated. High-resolution mapping of these mRNA's by using S1 nuclease and exonuclease VII digestion of hybrids between them and 5' and 3' end-labeled DNA fragments from the region indicated that the major early mRNA contained no detectable splices, and about half of its 3' end was complementary to the 3' region of the very minor 2.6- to 2.8-kb mRNA's encoded on the opposite strand. These mRNA's also contained no detectable splices. The major late 2.7-kb mRNA was found to be a family made up of members with no detectable splices and members with variable-length (100 to 300 bases) segments spliced out very near (ca. 50 to 100 bases) the 5' end.

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  1. Aleström P., Akusjärvi G., Perricaudet M., Mathews M. B., Klessig D. F., Pettersson U. The gene for polypeptide IX of adenovirus type 2 and its unspliced messenger RNA. Cell. 1980 Mar;19(3):671–681. doi: 10.1016/s0092-8674(80)80044-4. [DOI] [PubMed] [Google Scholar]
  2. Alwine J. C., Kemp D. J., Stark G. R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350–5354. doi: 10.1073/pnas.74.12.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anderson K. P., Costa R. H., Holland L. E., Wagner E. K. Characterization of herpes simplex virus type 1 RNA present in the absence of de novo protein synthesis. J Virol. 1980 Apr;34(1):9–27. doi: 10.1128/jvi.34.1.9-27.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Anderson K. P., Frink R. J., Devi G. B., Gaylord B. H., Costa R. H., Wagner E. K. Detailed characterization of the mRNA mapping in the HindIII fragment K region of the herpes simplex virus type 1 genome. J Virol. 1981 Mar;37(3):1011–1027. doi: 10.1128/jvi.37.3.1011-1027.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Anderson K. P., Holland L. E., Gaylord B. H., Wagner E. K. Isolation and translation of mRNA encoded by a specific region of the herpes simplex virus type 1 genome. J Virol. 1980 Feb;33(2):749–759. doi: 10.1128/jvi.33.2.749-759.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Anderson K. P., Stringer J. R., Holland L. E., Wagner E. K. Isolation and localization of herpes simplex virus type 1 mRNA. J Virol. 1979 Jun;30(3):805–820. doi: 10.1128/jvi.30.3.805-820.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bachenheimer S. L., Roizman B. Ribonucleic acid synthesis in cells infected with herpes simplex virus. VI. Polyadenylic acid sequences in viral messenger ribonucleic acid. J Virol. 1972 Oct;10(4):875–879. doi: 10.1128/jvi.10.4.875-879.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bailey J. M., Davidson N. Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Anal Biochem. 1976 Jan;70(1):75–85. doi: 10.1016/s0003-2697(76)80049-8. [DOI] [PubMed] [Google Scholar]
  9. Bartkoski M., Roizman B. RNA synthesis in cells infected with herpes simple virus. XIII. Differences in the methylation patterns of viral RNA during the reproductive cycle. J Virol. 1976 Dec;20(3):583–588. doi: 10.1128/jvi.20.3.583-588.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Bedbrook J. R., Kolodner R., Bogorad L. Zea mays chloroplast ribosomal RNA genes are part of a 22,000 base pair inverted repeat. Cell. 1977 Aug;11(4):739–749. doi: 10.1016/0092-8674(77)90288-4. [DOI] [PubMed] [Google Scholar]
  11. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  12. Berk A. J., Sharp P. A. Spliced early mRNAs of simian virus 40. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1274–1278. doi: 10.1073/pnas.75.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Berk A. J., Sharp P. A. Structure of the adenovirus 2 early mRNAs. Cell. 1978 Jul;14(3):695–711. doi: 10.1016/0092-8674(78)90252-0. [DOI] [PubMed] [Google Scholar]
  14. Clements J. B., McLauchlan J., McGeoch D. J. Orientation of herpes simplex virus type 1 immediate early mRNA's. Nucleic Acids Res. 1979 Sep 11;7(1):77–91. doi: 10.1093/nar/7.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Clements J. B., Watson R. J., Wilkie N. M. Temporal regulation of herpes simplex virus type 1 transcription: location of transcripts on the viral genome. Cell. 1977 Sep;12(1):275–285. doi: 10.1016/0092-8674(77)90205-7. [DOI] [PubMed] [Google Scholar]
  16. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Costa R. H., Devi B. G., Anderson K. P., Gaylord B. H., Wagner E. K. Characterization of a major late herpes simplex virus type 1 mRNA. J Virol. 1981 May;38(2):483–496. doi: 10.1128/jvi.38.2.483-496.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Courtney R. J., Schaffer P. A., Powell K. L. Synthesis of virus-specific polypaptides by temperature-sensitive mutants of herpes simplex virus type 1. Virology. 1976 Dec;75(2):306–318. doi: 10.1016/0042-6822(76)90030-1. [DOI] [PubMed] [Google Scholar]
  20. Cremer K., Bodemer M., Summers W. C. Characterization of the mRNA for herpes simplex virus thymidine kinase by cell-free synthesis of active enzyme. Nucleic Acids Res. 1978 Jul;5(7):2333–2344. doi: 10.1093/nar/5.7.2333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  22. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hamer D. H., Leder P. Splicing and the formation of stable RNA. Cell. 1979 Dec;18(4):1299–1302. doi: 10.1016/0092-8674(79)90240-x. [DOI] [PubMed] [Google Scholar]
  24. Hayward G. S., Jacob R. J., Wadsworth S. C., Roizman B. Anatomy of herpes simplex virus DNA: evidence for four populations of molecules that differ in the relative orientations of their long and short components. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4243–4247. doi: 10.1073/pnas.72.11.4243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Holland L. E., Anderson K. P., Shipman C., Jr, Wagner E. K. Viral DNA synthesis is required for the efficient expression of specific herpes simplex virus type 1 mRNA species. Virology. 1980 Feb;101(1):10–24. doi: 10.1016/0042-6822(80)90479-1. [DOI] [PubMed] [Google Scholar]
  26. Holland L. E., Anderson K. P., Stringer J. R., Wagner E. K. Isolation and localization of herpes simplex virus type 1 mRNA abundant before viral DNA synthesis. J Virol. 1979 Aug;31(2):447–462. doi: 10.1128/jvi.31.2.447-462.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Honess R. W., Roizman B. Regulation of herpesvirus macromolecular synthesis. I. Cascade regulation of the synthesis of three groups of viral proteins. J Virol. 1974 Jul;14(1):8–19. doi: 10.1128/jvi.14.1.8-19.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Jones P. C., Hayward G. S., Roizman B. Anatomy of herpes simplex virus DNA VII. alpha-RNA is homologous to noncontiguous sites in both the L and S components of viral DNA. J Virol. 1977 Jan;21(1):268–276. doi: 10.1128/jvi.21.1.268-276.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Jones P. C., Roizman B. Regulation of herpesvirus macromolecular synthesis. VIII. The transcription program consists of three phases during which both extent of transcription and accumulation of RNA in the cytoplasm are regulated. J Virol. 1979 Aug;31(2):299–314. doi: 10.1128/jvi.31.2.299-314.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kozak M., Roizman B. Regulation of herpesvirus macromolecular synthesis: nuclear retention of nontranslated viral RNA sequences. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4322–4326. doi: 10.1073/pnas.71.11.4322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  32. Marsden H. S., Crombie I. K., Subak-Sharpe J. H. Control of protein synthesis in herpesvirus-infected cells: analysis of the polypeptides induced by wild type and sixteen temperature-sensitive mutants of HSV strain 17. J Gen Virol. 1976 Jun;31(3):347–372. doi: 10.1099/0022-1317-31-3-347. [DOI] [PubMed] [Google Scholar]
  33. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  34. McKnight S. L. The nucleotide sequence and transcript map of the herpes simplex virus thymidine kinase gene. Nucleic Acids Res. 1980 Dec 20;8(24):5949–5964. doi: 10.1093/nar/8.24.5949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Moss B., Gershowitz A., Stringer J. R., Holland L. E., Wagner E. K. 5'-Terminal and internal methylated nucleosides in herpes simplex virus type 1 mRNA. J Virol. 1977 Aug;23(2):234–239. doi: 10.1128/jvi.23.2.234-239.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Murray B. K., Benyesh-Melnick M., Biswal N. Early and late viral-specific polyribosomal RNA in herpes virus-1 and -2-infected rabbit kidney cells. Biochim Biophys Acta. 1974 Aug 29;361(2):209–220. doi: 10.1016/0005-2787(74)90348-7. [DOI] [PubMed] [Google Scholar]
  38. Palmiter R. D. Magnesium precipitation of ribonucleoprotein complexes. Expedient techniques for the isolation of undergraded polysomes and messenger ribonucleic acid. Biochemistry. 1974 Aug 13;13(17):3606–3615. doi: 10.1021/bi00714a032. [DOI] [PubMed] [Google Scholar]
  39. Powell K. L., Purifoy D. J., Courtney R. J. The synthesis of herpes simplex virus proteins in the absence of virus DNA synthesis. Biochem Biophys Res Commun. 1975 Sep 2;66(1):262–271. doi: 10.1016/s0006-291x(75)80323-8. [DOI] [PubMed] [Google Scholar]
  40. Preston C. M. Abnormal properties of an immediate early polypeptide in cells infected with the herpes simplex virus type 1 mutant tsK. J Virol. 1979 Nov;32(2):357–369. doi: 10.1128/jvi.32.2.357-369.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Preston C. M. Cell-free synthesis of herpes simplex virus-coded pyrimidine deoxyribonucleoside kinase enzyme. J Virol. 1977 Sep;23(3):455–460. doi: 10.1128/jvi.23.3.455-460.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Rakusanova T., Ben-Porat T., Himeno M., Kaplan A. S. Early functions of the genome of herpesvirus. I. Characterization of the RNA synthesized in cycloheximide-treated, infected cells. Virology. 1971 Dec;46(3):877–889. doi: 10.1016/0042-6822(71)90088-2. [DOI] [PubMed] [Google Scholar]
  43. Roizman B. The structure and isomerization of herpes simplex virus genomes. Cell. 1979 Mar;16(3):481–494. doi: 10.1016/0092-8674(79)90023-0. [DOI] [PubMed] [Google Scholar]
  44. Silvertien S., Millette R., Jones P., Roizman B. RNA synthesis in cells infected with herpes simplex virus. XII. Sequence complexity and properties of RNA differing in extent of adenylation. J Virol. 1976 Jun;18(3):977–991. doi: 10.1128/jvi.18.3.977-991.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Skare J., Summers W. C. Structure and function of herpesvirus genomes. II. EcoRl, Sbal, and HindIII endonuclease cleavage sites on herpes simplex virus. Virology. 1977 Feb;76(2):581–595. doi: 10.1016/0042-6822(77)90240-9. [DOI] [PubMed] [Google Scholar]
  46. Stringer J. R., Holland L. E., Swanstrom R. I., Pivo K., Wagner E. K. Quantitation of herpes simplex virus type 1 RNA in infected HeLa cells. J Virol. 1977 Mar;21(3):889–901. doi: 10.1128/jvi.21.3.889-901.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Stringer J. R., Holland L. E., Wagner E. K. Mapping early transcripts of herpes simplex virus type 1 by electron microscopy. J Virol. 1978 Jul;27(1):56–73. doi: 10.1128/jvi.27.1.56-73.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Swanstrom R. I., Pivo K., Wagner E. K. Restricted transcription of the herpes simplex virus genome occurring early after infection and in the presence of metabolic inhibitors. Virology. 1975 Jul;66(1):140–150. doi: 10.1016/0042-6822(75)90185-3. [DOI] [PubMed] [Google Scholar]
  49. Swanstrom R. I., Wagner E. K. Regulation of synthesis of herpes simplex type 1 virus mRNA during productive infection. Virology. 1974 Aug;60(2):522–533. doi: 10.1016/0042-6822(74)90346-8. [DOI] [PubMed] [Google Scholar]
  50. Tanaka T., Weisblum B. Construction of a colicin E1-R factor composite plasmid in vitro: means for amplification of deoxyribonucleic acid. J Bacteriol. 1975 Jan;121(1):354–362. doi: 10.1128/jb.121.1.354-362.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Taylor J. M., Illmensee R., Summers J. Efficeint transcription of RNA into DNA by avian sarcoma virus polymerase. Biochim Biophys Acta. 1976 Sep 6;442(3):324–330. doi: 10.1016/0005-2787(76)90307-5. [DOI] [PubMed] [Google Scholar]
  52. Thomas M., Davis R. W. Studies on the cleavage of bacteriophage lambda DNA with EcoRI Restriction endonuclease. J Mol Biol. 1975 Jan 25;91(3):315–328. doi: 10.1016/0022-2836(75)90383-6. [DOI] [PubMed] [Google Scholar]
  53. Wadsworth S., Hayward G. S., Roizman B. Anatomy of herpes simplex virus DNA. V. Terminally repetitive sequences. J Virol. 1976 Feb;17(2):503–512. doi: 10.1128/jvi.17.2.503-512.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Wagner E. K. Evidence for transcriptional control of the herpes simplex virus genome in infected human cells. Virology. 1972 Feb;47(2):502–506. doi: 10.1016/0042-6822(72)90289-9. [DOI] [PubMed] [Google Scholar]
  55. Wagner E. K., Roizman B. RNA synthesis in cells infected with herpes simplex virus. II. Evidence that a class of viral mRNA is derived from a high molecular weight precursor synthesized in the nucleus. Proc Natl Acad Sci U S A. 1969 Oct;64(2):626–633. doi: 10.1073/pnas.64.2.626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Wagner E. K., Swanstrom R. I., Stafford M. G. Transcription of the herpes simplex virus genome in human cells. J Virol. 1972 Oct;10(4):675–682. doi: 10.1128/jvi.10.4.675-682.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Ward R. L., Stevens J. G. Effect of cytosine arabinoside on viral-specific protein synthesis in cells infected with herpes simplex virus. J Virol. 1975 Jan;15(1):71–80. doi: 10.1128/jvi.15.1.71-80.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Watson R. J., Preston C. M., Clements J. B. Separation and characterization of herpes simplex virus type 1 immediate-early mRNA's. J Virol. 1979 Jul;31(1):42–52. doi: 10.1128/jvi.31.1.42-52.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Watson R. J., Sullivan M., Vande Woude G. F. Structures of two spliced herpes simplex virus type 1 immediate-early mRNA's which map at the junctions of the unique and reiterated regions of the virus DNA S component. J Virol. 1981 Jan;37(1):431–444. doi: 10.1128/jvi.37.1.431-444.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Wellauer P. K., Dawid I. B. Secondary structure maps of RNA: processing of HeLa ribosomal RNA. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2827–2831. doi: 10.1073/pnas.70.10.2827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Wilkie N. M., Cortini R. Sequence arrangement in herpes simplex virus type 1 DNA: identification of terminal fragments in restriction endonuclease digests and evidence for inversions in redundant and unique sequences. J Virol. 1976 Oct;20(1):211–221. doi: 10.1128/jvi.20.1.211-221.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Wilkie N. M. Physical maps for Herpes simplex virus type 1 DNA for restriction endonucleases Hind III, Hpa-1, and X. bad. J Virol. 1976 Oct;20(1):222–233. doi: 10.1128/jvi.20.1.222-233.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]

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