Abstract
The role of putative promoter or activator sequences downstream of the herpes simplex virus type 2 latency-associated transcript (LAT) promoter and upstream of the LAT intron was investigated in vivo by constructing and evaluating mutant viruses with deletions in this region. The deletion of LAT promoter sequences upstream of the primary LAT transcript reduced levels of LAT expression during productive infections, compared with the LAT expression level of wild-type virus, and abolished LAT expression during latency. The deletion of the putative downstream regulatory elements reduced but did not eliminate LAT expression during productive and latent infections. The deletion of both regions almost completely eliminated acute LAT transcription, although additional acute LAT-region transcription directed by sequences upstream of either region was detected by reverse transcriptase PCR. The deletion of the downstream elements did not influence the ability of the virus to reactivate from latently infected guinea pigs relative to the ability of the wild-type virus to reactivate; thus, decreased LAT expression did not affect the frequency of recurrence. The deletion of both regions did not affect the ability of the virus to establish latency. We conclude that downstream regulatory elements are necessary for maximal acute LAT expression but do not constitute an independent promoter during latency and do not play an obvious role in the establishment of our reactivation from latency.
Full Text
The Full Text of this article is available as a PDF (266.6 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Batchelor A. H., O'Hare P. Regulation and cell-type-specific activity of a promoter located upstream of the latency-associated transcript of herpes simplex virus type 1. J Virol. 1990 Jul;64(7):3269–3279. doi: 10.1128/jvi.64.7.3269-3279.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bloom D. C., Devi-Rao G. B., Hill J. M., Stevens J. G., Wagner E. K. Molecular analysis of herpes simplex virus type 1 during epinephrine-induced reactivation of latently infected rabbits in vivo. J Virol. 1994 Mar;68(3):1283–1292. doi: 10.1128/jvi.68.3.1283-1292.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bourne N., Stanberry L. R., Connelly B. L., Kurawadwala J., Straus S. E., Krause P. R. Quantity of latency-associated transcript produced by herpes simplex virus is not predictive of the frequency of experimental recurrent genital herpes. J Infect Dis. 1994 May;169(5):1084–1087. doi: 10.1093/infdis/169.5.1084. [DOI] [PubMed] [Google Scholar]
- Corey L., Spear P. G. Infections with herpes simplex viruses (1). N Engl J Med. 1986 Mar 13;314(11):686–691. doi: 10.1056/NEJM198603133141105. [DOI] [PubMed] [Google Scholar]
- 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]
- Croen K. D., Ostrove J. M., Dragovic L., Straus S. E. Characterization of herpes simplex virus type 2 latency-associated transcription in human sacral ganglia and in cell culture. J Infect Dis. 1991 Jan;163(1):23–28. doi: 10.1093/infdis/163.1.23. [DOI] [PubMed] [Google Scholar]
- 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]
- Dobson A. T., Margolis T. P., Gomes W. A., Feldman L. T. In vivo deletion analysis of the herpes simplex virus type 1 latency-associated transcript promoter. J Virol. 1995 Apr;69(4):2264–2270. doi: 10.1128/jvi.69.4.2264-2270.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Ecob-Prince M. S., Rixon F. J., Preston C. M., Hassan K., Kennedy P. G. Reactivation in vivo and in vitro of herpes simplex virus from mouse dorsal root ganglia which contain different levels of latency-associated transcripts. J Gen Virol. 1993 Jun;74(Pt 6):995–1002. doi: 10.1099/0022-1317-74-6-995. [DOI] [PubMed] [Google Scholar]
- Farrell M. J., Dobson A. T., Feldman L. T. Herpes simplex virus latency-associated transcript is a stable intron. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):790–794. doi: 10.1073/pnas.88.3.790. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goins W. F., Sternberg L. R., Croen K. D., Krause P. R., Hendricks R. L., Fink D. J., Straus S. E., Levine M., Glorioso J. C. A novel latency-active promoter is contained within the herpes simplex virus type 1 UL flanking repeats. J Virol. 1994 Apr;68(4):2239–2252. doi: 10.1128/jvi.68.4.2239-2252.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hill J. M., Sedarati F., Javier R. T., Wagner E. K., Stevens J. G. Herpes simplex virus latent phase transcription facilitates in vivo reactivation. Virology. 1990 Jan;174(1):117–125. doi: 10.1016/0042-6822(90)90060-5. [DOI] [PubMed] [Google Scholar]
- 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]
- Kolluri R., Torrey T. A., Kinniburgh A. J. A CT promoter element binding protein: definition of a double-strand and a novel single-strand DNA binding motif. Nucleic Acids Res. 1992 Jan 11;20(1):111–116. doi: 10.1093/nar/20.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Krause P. R., Ostrove J. M., Straus S. E. The nucleotide sequence, 5' end, promoter domain, and kinetics of expression of the gene encoding the herpes simplex virus type 2 latency-associated transcript. J Virol. 1991 Oct;65(10):5619–5623. doi: 10.1128/jvi.65.10.5619-5623.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Krause P. R., Stanberry L. R., Bourne N., Connelly B., Kurawadwala J. F., Patel A., Straus S. E. Expression of the herpes simplex virus type 2 latency-associated transcript enhances spontaneous reactivation of genital herpes in latently infected guinea pigs. J Exp Med. 1995 Jan 1;181(1):297–306. doi: 10.1084/jem.181.1.297. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lafferty W. E., Coombs R. W., Benedetti J., Critchlow C., Corey L. Recurrences after oral and genital herpes simplex virus infection. Influence of site of infection and viral type. N Engl J Med. 1987 Jun 4;316(23):1444–1449. doi: 10.1056/NEJM198706043162304. [DOI] [PubMed] [Google Scholar]
- Leib D. A., Bogard C. L., Kosz-Vnenchak M., Hicks K. A., Coen D. M., Knipe D. M., Schaffer P. A. A deletion mutant of the latency-associated transcript of herpes simplex virus type 1 reactivates from the latent state with reduced frequency. J Virol. 1989 Jul;63(7):2893–2900. doi: 10.1128/jvi.63.7.2893-2900.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lokensgard J. R., Bloom D. C., Dobson A. T., Feldman L. T. Long-term promoter activity during herpes simplex virus latency. J Virol. 1994 Nov;68(11):7148–7158. doi: 10.1128/jvi.68.11.7148-7158.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maggioncalda J., Mehta A., Fraser N. W., Block T. M. Analysis of a herpes simplex virus type 1 LAT mutant with a deletion between the putative promoter and the 5' end of the 2.0-kilobase transcript. J Virol. 1994 Dec;68(12):7816–7824. doi: 10.1128/jvi.68.12.7816-7824.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McGeoch D. J., Cunningham C., McIntyre G., Dolan A. Comparative sequence analysis of the long repeat regions and adjoining parts of the long unique regions in the genomes of herpes simplex viruses types 1 and 2. J Gen Virol. 1991 Dec;72(Pt 12):3057–3075. doi: 10.1099/0022-1317-72-12-3057. [DOI] [PubMed] [Google Scholar]
- Mitchell W. J., Deshmane S. L., Dolan A., McGeoch D. J., Fraser N. W. Characterization of herpes simplex virus type 2 transcription during latent infection of mouse trigeminal ganglia. J Virol. 1990 Nov;64(11):5342–5348. doi: 10.1128/jvi.64.11.5342-5348.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nicosia M., Deshmane S. L., Zabolotny J. M., Valyi-Nagy T., Fraser N. W. Herpes simplex virus type 1 latency-associated transcript (LAT) promoter deletion mutants can express a 2-kilobase transcript mapping to the LAT region. J Virol. 1993 Dec;67(12):7276–7283. doi: 10.1128/jvi.67.12.7276-7283.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rader K. A., Ackland-Berglund C. E., Miller J. K., Pepose J. S., Leib D. A. In vivo characterization of site-directed mutations in the promoter of the herpes simplex virus type 1 latency-associated transcripts. J Gen Virol. 1993 Sep;74(Pt 9):1859–1869. doi: 10.1099/0022-1317-74-9-1859. [DOI] [PubMed] [Google Scholar]
- Sawtell N. M., Thompson R. L. Herpes simplex virus type 1 latency-associated transcription unit promotes anatomical site-dependent establishment and reactivation from latency. J Virol. 1992 Apr;66(4):2157–2169. doi: 10.1128/jvi.66.4.2157-2169.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Singh J., Wagner E. K. Transcriptional analysis of the herpes simplex virus type 1 region containing the TRL/UL junction. Virology. 1993 Sep;196(1):220–231. doi: 10.1006/viro.1993.1470. [DOI] [PubMed] [Google Scholar]
- 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]
- Stanberry L. R., Kern E. R., Richards J. T., Abbott T. M., Overall J. C., Jr Genital herpes in guinea pigs: pathogenesis of the primary infection and description of recurrent disease. J Infect Dis. 1982 Sep;146(3):397–404. doi: 10.1093/infdis/146.3.397. [DOI] [PubMed] [Google Scholar]
- Stanberry L. R., Kern E. R., Richards J. T., Overall J. C., Jr Recurrent genital herpes simplex virus infection in guinea pigs. Intervirology. 1985;24(4):226–231. doi: 10.1159/000149647. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Trousdale M. D., Steiner I., Spivack J. G., Deshmane S. L., Brown S. M., MacLean A. R., Subak-Sharpe J. H., Fraser N. W. In vivo and in vitro reactivation impairment of a herpes simplex virus type 1 latency-associated transcript variant in a rabbit eye model. J Virol. 1991 Dec;65(12):6989–6993. doi: 10.1128/jvi.65.12.6989-6993.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Wang K., Krause P. R., Straus S. E. Analysis of the promoter and cis-acting elements regulating expression of herpes simplex virus type 2 latency-associated transcripts. J Virol. 1995 May;69(5):2873–2880. doi: 10.1128/jvi.69.5.2873-2880.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Zwaagstra J. C., Ghiasi H., Nesburn A. B., Wechsler S. L. Identification of a major regulatory sequence in the latency associated transcript (LAT) promoter of herpes simplex virus type 1 (HSV-1). Virology. 1991 May;182(1):287–297. doi: 10.1016/0042-6822(91)90672-x. [DOI] [PubMed] [Google Scholar]
- Zwaagstra J. C., Ghiasi H., Slanina S. M., Nesburn A. B., Wheatley S. C., Lillycrop K., Wood J., Latchman D. S., Patel K., Wechsler S. L. Activity of herpes simplex virus type 1 latency-associated transcript (LAT) promoter in neuron-derived cells: evidence for neuron specificity and for a large LAT transcript. J Virol. 1990 Oct;64(10):5019–5028. doi: 10.1128/jvi.64.10.5019-5028.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]