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. 1990 Jul;64(7):3269–3279. doi: 10.1128/jvi.64.7.3269-3279.1990

Regulation and cell-type-specific activity of a promoter located upstream of the latency-associated transcript of herpes simplex virus type 1.

A H Batchelor 1, P O'Hare 1
PMCID: PMC249552  PMID: 2161941

Abstract

To identify promoter regions which control expression of the latency-associated transcript (LAT) of herpes simplex virus type 1 (HSV-1), we constructed a series of recombinant vectors in which various sequences upstream of LAT were linked to the chloramphenicol acetyltransferase gene and tested for expression efficiency by transfection into tissue culture cells. In HeLa cells no activity was observed from the region (-250 to +201) immediately surrounding the nominal 5' end of LAT, but high levels of activity were observed by using different fragments within the region -1267 to -594. This promoter activity was largely contained within the 140-base-pair region from -797 to -658 and was 20- to 50-fold stronger than typical HSV delayed-early promoters and at least as strong as the activity from the simian virus 40 (SV40) enhancer-promoter region or the HSV immediate-early 110,000-Mr (IE110K) promoter region. In human neuroblastoma cells (IMR-32), there was a dramatic switch in relative activities in favor of the LAT promoter, so that it was 45- and 200-fold stronger than the IE110K and SV40 constructs, respectively. Furthermore, optimal activity in the neuroblastoma cells required sequences within the region -1267 to -797. This region had little effect on activity in HeLa cells. We also show that the LAT promoter activity was very efficiently repressed by the IE175K protein. From internal deletion analysis, the site of repression was located within a 55-base-pair region just downstream of a potential TATA box. This region exhibited a high degree of homology with the IE175K cap site and may be a binding site for the IE175K protein.

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Selected References

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

  1. Boshart M., Weber F., Jahn G., Dorsch-Häsler K., Fleckenstein B., Schaffner W. A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus. Cell. 1985 Jun;41(2):521–530. doi: 10.1016/s0092-8674(85)80025-8. [DOI] [PubMed] [Google Scholar]
  2. Bray S. J., Johnson W. A., Hirsh J., Heberlein U., Tjian R. A cis-acting element and associated binding factor required for CNS expression of the Drosophila melanogaster dopa decarboxylase gene. EMBO J. 1988 Jan;7(1):177–188. doi: 10.1002/j.1460-2075.1988.tb02798.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Coen D. M., Weinheimer S. P., McKnight S. L. A genetic approach to promoter recognition during trans induction of viral gene expression. Science. 1986 Oct 3;234(4772):53–59. doi: 10.1126/science.3018926. [DOI] [PubMed] [Google Scholar]
  5. Costa R. H., Draper K. G., Devi-Rao G., Thompson R. L., Wagner E. K. Virus-induced modification of the host cell is required for expression of the bacterial chloramphenicol acetyltransferase gene controlled by a late herpes simplex virus promoter (VP5). J Virol. 1985 Oct;56(1):19–30. doi: 10.1128/jvi.56.1.19-30.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Croen K. D., Ostrove J. M., Dragovic L. J., Smialek J. E., Straus S. E. Latent herpes simplex virus in human trigeminal ganglia. Detection of an immediate early gene "anti-sense" transcript by in situ hybridization. N Engl J Med. 1987 Dec 3;317(23):1427–1432. doi: 10.1056/NEJM198712033172302. [DOI] [PubMed] [Google Scholar]
  7. Danciger E., Mettling C., Vidal M., Morris R., Margolis F. Olfactory marker protein gene: its structure and olfactory neuron-specific expression in transgenic mice. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8565–8569. doi: 10.1073/pnas.86.21.8565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Deatly A. M., Spivack J. G., Lavi E., Fraser N. W. RNA from an immediate early region of the type 1 herpes simplex virus genome is present in the trigeminal ganglia of latently infected mice. Proc Natl Acad Sci U S A. 1987 May;84(10):3204–3208. doi: 10.1073/pnas.84.10.3204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dobson A. T., Sederati F., Devi-Rao G., Flanagan W. M., Farrell M. J., Stevens J. G., Wagner E. K., Feldman L. T. Identification of the latency-associated transcript promoter by expression of rabbit beta-globin mRNA in mouse sensory nerve ganglia latently infected with a recombinant herpes simplex virus. J Virol. 1989 Sep;63(9):3844–3851. doi: 10.1128/jvi.63.9.3844-3851.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Faber S. W., Wilcox K. W. Association of herpes simplex virus regulatory protein ICP4 with sequences spanning the ICP4 gene transcription initiation site. Nucleic Acids Res. 1988 Jan 25;16(2):555–570. doi: 10.1093/nar/16.2.555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Galloway D. A., Fenoglio C. M., McDougall J. K. Limited transcription of the herpes simplex virus genome when latent in human sensory ganglia. J Virol. 1982 Feb;41(2):686–691. doi: 10.1128/jvi.41.2.686-691.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gelman I. H., Silverstein S. Identification of immediate early genes from herpes simplex virus that transactivate the virus thymidine kinase gene. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5265–5269. doi: 10.1073/pnas.82.16.5265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gordon Y. J., Johnson B., Romanowski E., Araullo-Cruz T. RNA complementary to herpes simplex virus type 1 ICP0 gene demonstrated in neurons of human trigeminal ganglia. J Virol. 1988 May;62(5):1832–1835. doi: 10.1128/jvi.62.5.1832-1835.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  16. Green M. T., Courtney R. J., Dunkel E. C. Detection of an immediate early herpes simplex virus type 1 polypeptide in trigeminal ganglia from latently infected animals. Infect Immun. 1981 Dec;34(3):987–992. doi: 10.1128/iai.34.3.987-992.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Honess R. W., Watson D. H. Unity and diversity in the herpesviruses. J Gen Virol. 1977 Oct;37(1):15–37. doi: 10.1099/0022-1317-37-1-15. [DOI] [PubMed] [Google Scholar]
  18. Javier R. T., Stevens J. G., Dissette V. B., Wagner E. K. A herpes simplex virus transcript abundant in latently infected neurons is dispensable for establishment of the latent state. Virology. 1988 Sep;166(1):254–257. doi: 10.1016/0042-6822(88)90169-9. [DOI] [PubMed] [Google Scholar]
  19. Johnson W. A., McCormick C. A., Bray S. J., Hirsh J. A neuron-specific enhancer of the Drosophila dopa decarboxylase gene. Genes Dev. 1989 May;3(5):676–686. doi: 10.1101/gad.3.5.676. [DOI] [PubMed] [Google Scholar]
  20. Jones C., Delhon G., Bratanich A., Kutish G., Rock D. Analysis of the transcriptional promoter which regulates the latency-related transcript of bovine herpesvirus 1. J Virol. 1990 Mar;64(3):1164–1170. doi: 10.1128/jvi.64.3.1164-1170.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kenney S., Natarajan V., Strike D., Khoury G., Salzman N. P. JC virus enhancer-promoter active in human brain cells. Science. 1984 Dec 14;226(4680):1337–1339. doi: 10.1126/science.6095453. [DOI] [PubMed] [Google Scholar]
  22. Krause P. R., Croen K. D., Straus S. E., Ostrove J. M. Detection and preliminary characterization of herpes simplex virus type 1 transcripts in latently infected human trigeminal ganglia. J Virol. 1988 Dec;62(12):4819–4823. doi: 10.1128/jvi.62.12.4819-4823.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Levine M., Goldin A. L., Glorioso J. C. Persistence of herpes simplex virus genes in cells of neuronal origin. J Virol. 1980 Jul;35(1):203–210. doi: 10.1128/jvi.35.1.203-210.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mackem S., Roizman B. Differentiation between alpha promoter and regulator regions of herpes simplex virus 1: the functional domains and sequence of a movable alpha regulator. Proc Natl Acad Sci U S A. 1982 Aug;79(16):4917–4921. doi: 10.1073/pnas.79.16.4917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. McGeoch D. J., Dalrymple M. A., Davison A. J., Dolan A., Frame M. C., McNab D., Perry L. J., Scott J. E., Taylor P. The complete DNA sequence of the long unique region in the genome of herpes simplex virus type 1. J Gen Virol. 1988 Jul;69(Pt 7):1531–1574. doi: 10.1099/0022-1317-69-7-1531. [DOI] [PubMed] [Google Scholar]
  26. Muller M. T. Binding of the herpes simplex virus immediate-early gene product ICP4 to its own transcription start site. J Virol. 1987 Mar;61(3):858–865. doi: 10.1128/jvi.61.3.858-865.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nilheden E., Jeansson S., Vahlne A. Herpes simplex virus latency in a hyperresistant clone of mouse neuroblastoma (Cl300) cells. Arch Virol. 1985;83(3-4):319–325. doi: 10.1007/BF01309927. [DOI] [PubMed] [Google Scholar]
  28. O'Hare P., Goding C. R. Herpes simplex virus regulatory elements and the immunoglobulin octamer domain bind a common factor and are both targets for virion transactivation. Cell. 1988 Feb 12;52(3):435–445. doi: 10.1016/s0092-8674(88)80036-9. [DOI] [PubMed] [Google Scholar]
  29. O'Hare P., Hayward G. S. Comparison of upstream sequence requirements for positive and negative regulation of a herpes simplex virus immediate-early gene by three virus-encoded trans-acting factors. J Virol. 1987 Jan;61(1):190–199. doi: 10.1128/jvi.61.1.190-199.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. O'Hare P., Hayward G. S. Expression of recombinant genes containing herpes simplex virus delayed-early and immediate-early regulatory regions and trans activation by herpesvirus infection. J Virol. 1984 Nov;52(2):522–531. doi: 10.1128/jvi.52.2.522-531.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. O'Hare P., Hayward G. S. Three trans-acting regulatory proteins of herpes simplex virus modulate immediate-early gene expression in a pathway involving positive and negative feedback regulation. J Virol. 1985 Dec;56(3):723–733. doi: 10.1128/jvi.56.3.723-733.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Perry L. J., McGeoch D. J. The DNA sequences of the long repeat region and adjoining parts of the long unique region in the genome of herpes simplex virus type 1. J Gen Virol. 1988 Nov;69(Pt 11):2831–2846. doi: 10.1099/0022-1317-69-11-2831. [DOI] [PubMed] [Google Scholar]
  33. Perry L. J., Rixon F. J., Everett R. D., Frame M. C., McGeoch D. J. Characterization of the IE110 gene of herpes simplex virus type 1. J Gen Virol. 1986 Nov;67(Pt 11):2365–2380. doi: 10.1099/0022-1317-67-11-2365. [DOI] [PubMed] [Google Scholar]
  34. Price R. W. Herpes simplex virus latency: adaptation to the peripheral nervous system. II. Cancer Invest. 1985;3(4):389–403. doi: 10.3109/07357908509039799. [DOI] [PubMed] [Google Scholar]
  35. Puga A., Rosenthal J. D., Openshaw H., Notkins A. L. Herpes simplex virus DNA and mRNA sequences in acutely and chronically infected trigeminal ganglia of mice. Virology. 1978 Aug;89(1):102–111. doi: 10.1016/0042-6822(78)90044-2. [DOI] [PubMed] [Google Scholar]
  36. Roberts M. S., Boundy A., O'Hare P., Pizzorno M. C., Ciufo D. M., Hayward G. S. Direct correlation between a negative autoregulatory response element at the cap site of the herpes simplex virus type 1 IE175 (alpha 4) promoter and a specific binding site for the IE175 (ICP4) protein. J Virol. 1988 Nov;62(11):4307–4320. doi: 10.1128/jvi.62.11.4307-4320.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rock D. L., Nesburn A. B., Ghiasi H., Ong J., Lewis T. L., Lokensgard J. R., Wechsler S. L. Detection of latency-related viral RNAs in trigeminal ganglia of rabbits latently infected with herpes simplex virus type 1. J Virol. 1987 Dec;61(12):3820–3826. doi: 10.1128/jvi.61.12.3820-3826.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Russell J., Preston C. M. An in vitro latency system for herpes simplex virus type 2. J Gen Virol. 1986 Feb;67(Pt 2):397–403. doi: 10.1099/0022-1317-67-2-397. [DOI] [PubMed] [Google Scholar]
  39. Spivack J. G., Fraser N. W. Detection of herpes simplex virus type 1 transcripts during latent infection in mice. J Virol. 1987 Dec;61(12):3841–3847. doi: 10.1128/jvi.61.12.3841-3847.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Spivack J. G., Fraser N. W. Expression of herpes simplex virus type 1 latency-associated transcripts in the trigeminal ganglia of mice during acute infection and reactivation of latent infection. J Virol. 1988 May;62(5):1479–1485. doi: 10.1128/jvi.62.5.1479-1485.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Steiner I., Spivack J. G., Lirette R. P., Brown S. M., MacLean A. R., Subak-Sharpe J. H., Fraser N. W. Herpes simplex virus type 1 latency-associated transcripts are evidently not essential for latent infection. EMBO J. 1989 Feb;8(2):505–511. doi: 10.1002/j.1460-2075.1989.tb03404.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Steiner I., Spivack J. G., O'Boyle D. R., 2nd, Lavi E., Fraser N. W. Latent herpes simplex virus type 1 transcription in human trigeminal ganglia. J Virol. 1988 Sep;62(9):3493–3496. doi: 10.1128/jvi.62.9.3493-3496.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Stevens J. G., Wagner E. K., Devi-Rao G. B., Cook M. L., Feldman L. T. RNA complementary to a herpesvirus alpha gene mRNA is prominent in latently infected neurons. Science. 1987 Feb 27;235(4792):1056–1059. doi: 10.1126/science.2434993. [DOI] [PubMed] [Google Scholar]
  44. Stinski M. F., Roehr T. J. Activation of the major immediate early gene of human cytomegalovirus by cis-acting elements in the promoter-regulatory sequence and by virus-specific trans-acting components. J Virol. 1985 Aug;55(2):431–441. doi: 10.1128/jvi.55.2.431-441.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tumilowicz J. J., Nichols W. W., Cholon J. J., Greene A. E. Definition of a continuous human cell line derived from neuroblastoma. Cancer Res. 1970 Aug;30(8):2110–2118. [PubMed] [Google Scholar]
  46. Wagner E. K., Devi-Rao G., Feldman L. T., Dobson A. T., Zhang Y. F., Flanagan W. M., Stevens J. G. Physical characterization of the herpes simplex virus latency-associated transcript in neurons. J Virol. 1988 Apr;62(4):1194–1202. doi: 10.1128/jvi.62.4.1194-1202.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wagner E. K., Flanagan W. M., Devi-Rao G., Zhang Y. F., Hill J. M., Anderson K. P., Stevens J. G. The herpes simplex virus latency-associated transcript is spliced during the latent phase of infection. J Virol. 1988 Dec;62(12):4577–4585. doi: 10.1128/jvi.62.12.4577-4585.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Wechsler S. L., Nesburn A. B., Watson R., Slanina S. M., Ghiasi H. Fine mapping of the latency-related gene of herpes simplex virus type 1: alternative splicing produces distinct latency-related RNAs containing open reading frames. J Virol. 1988 Nov;62(11):4051–4058. doi: 10.1128/jvi.62.11.4051-4058.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wechsler S. L., Nesburn A. B., Watson R., Slanina S., Ghiasi H. Fine mapping of the major latency-related RNA of herpes simplex virus type 1 in humans. J Gen Virol. 1988 Dec;69(Pt 12):3101–3106. doi: 10.1099/0022-1317-69-12-3101. [DOI] [PubMed] [Google Scholar]
  50. Wechsler S. L., Nesburn A. B., Zwaagstra J., Ghiasi H. Sequence of the latency-related gene of herpes simplex virus type 1. Virology. 1989 Jan;168(1):168–172. doi: 10.1016/0042-6822(89)90416-9. [DOI] [PubMed] [Google Scholar]
  51. Weeks D. L., Jones N. C. Adenovirus E3-early promoter: sequences required for activation by E1A. Nucleic Acids Res. 1985 Jul 25;13(14):5389–5402. doi: 10.1093/nar/13.14.5389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Wigdahl B. L., Ziegler R. J., Sneve M., Rapp F. Herpes simplex virus latency and reactivation in isolated rat sensory neurons. Virology. 1983 May;127(1):159–167. doi: 10.1016/0042-6822(83)90380-x. [DOI] [PubMed] [Google Scholar]
  53. Wilcox C. L., Johnson E. M., Jr Characterization of nerve growth factor-dependent herpes simplex virus latency in neurons in vitro. J Virol. 1988 Feb;62(2):393–399. doi: 10.1128/jvi.62.2.393-399.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Yamamoto H., Walz M. A., Notkins A. L. Viral-specific thymidine kinase in sensory ganglia of mice infected with herpes simplex virus. Virology. 1977 Feb;76(2):866–869. doi: 10.1016/0042-6822(77)90267-7. [DOI] [PubMed] [Google Scholar]
  55. Yura Y., Terashima K., Iga H., Yanagawa T., Yoshida H., Hayashi Y., Sato M. A latent infection of herpes simplex virus type 2 in a human neuroblastoma cell line IMR-32. Arch Virol. 1986;90(3-4):249–260. doi: 10.1007/BF01317374. [DOI] [PubMed] [Google Scholar]
  56. Zwaagstra J., Ghiasi H., Nesburn A. B., Wechsler S. L. In vitro promoter activity associated with the latency-associated transcript gene of herpes simplex virus type 1. J Gen Virol. 1989 Aug;70(Pt 8):2163–2169. doi: 10.1099/0022-1317-70-8-2163. [DOI] [PubMed] [Google Scholar]

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