Effect of the transcription start region of the herpes simplex virus type 1 latency-associated transcript promoter on expression of productively infected neurons in vivo

Michael J. Farrell, Todd P. Margolis, William A. Gomes, Lawrence T. Feldman

Research output: Contribution to journalArticle

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Abstract

It has been previously reported that the latency-associated transcript (LAT) promoter contains a DNA sequence at the LAT transcription start site which resembles the ICP4 consensus DNA binding site and that this site allows ICP4-mediated downregulation of the LAT promoter in transient assays (A. H. Batchelor and P. O'Hare, J. Virol. 64:3269-3279, 1990). We have confirmed these data by showing that an ICP4-expressing plasmid will downregulate lacZ expression from a plasmid containing the LAT promoter and transcription start site (pJA1) and does not downregulate lacZ expression from a plasmid in which the start site has been mutagenized (pWAG15). To determine the role of the LAT transcription start site in regulating LAT promoter activity in the context of the virus, two recombinant viruses, KOS-1 and KOS-15, were studied. KOS-1 contains an 863-bp portion of the LAT promoter, including the LAT cap site, fused to the lacZ gene and inserted into the gC locus (T. P. Margolis, F. Sedarati, A. T. Dobson, L. T. Feldman, and J. G. Stevens, Virology 189:150-160, 1992). The second virus (KOS-15) was constructed in identical fashion, using plasmid pWAG-15, which is not downregulated by ICP4. Vero cells productively infected with KOS-15 produce 10-fold more β- galactosidase than do those infected with KOS-1. In murine dorsal root ganglia acutely infected with KOS-1, only 1.2% of dorsal root ganglion neurons that expressed viral antigen also expressed β-galactosidase. In contrast, in KOS-15-infected mice, β-galactosidase was detected in 18% of viral antigen-positive neurons. Similar findings were observed in trigeminal ganglia acutely infected with KOS-1 and KOS-15. Thus, the region encompassing the LAT transcription start site appears to play an important role in repression of the LAT promoter activity not only in vitro but also in acutely infected neurons in vivo. These results suggest that during productive infection with HSV-1, LAT expression is tightly regulated.

Original languageEnglish (US)
Pages (from-to)5337-5343
Number of pages7
JournalJournal of Virology
Volume68
Issue number9
StatePublished - Sep 1994
Externally publishedYes

Fingerprint

Human herpesvirus 1
Transcription Initiation Site
Human Herpesvirus 1
Galactosidases
Plasmids
Down-Regulation
transcription (genetics)
neurons
promoter regions
galactosidases
Neurons
Viral Antigens
plasmids
Spinal Ganglia
Viruses
viral antigens
Trigeminal Ganglion
viruses
Virology
Vero Cells

ASJC Scopus subject areas

  • Immunology

Cite this

Effect of the transcription start region of the herpes simplex virus type 1 latency-associated transcript promoter on expression of productively infected neurons in vivo. / Farrell, Michael J.; Margolis, Todd P.; Gomes, William A.; Feldman, Lawrence T.

In: Journal of Virology, Vol. 68, No. 9, 09.1994, p. 5337-5343.

Research output: Contribution to journalArticle

Farrell, Michael J. ; Margolis, Todd P. ; Gomes, William A. ; Feldman, Lawrence T. / Effect of the transcription start region of the herpes simplex virus type 1 latency-associated transcript promoter on expression of productively infected neurons in vivo. In: Journal of Virology. 1994 ; Vol. 68, No. 9. pp. 5337-5343.
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abstract = "It has been previously reported that the latency-associated transcript (LAT) promoter contains a DNA sequence at the LAT transcription start site which resembles the ICP4 consensus DNA binding site and that this site allows ICP4-mediated downregulation of the LAT promoter in transient assays (A. H. Batchelor and P. O'Hare, J. Virol. 64:3269-3279, 1990). We have confirmed these data by showing that an ICP4-expressing plasmid will downregulate lacZ expression from a plasmid containing the LAT promoter and transcription start site (pJA1) and does not downregulate lacZ expression from a plasmid in which the start site has been mutagenized (pWAG15). To determine the role of the LAT transcription start site in regulating LAT promoter activity in the context of the virus, two recombinant viruses, KOS-1 and KOS-15, were studied. KOS-1 contains an 863-bp portion of the LAT promoter, including the LAT cap site, fused to the lacZ gene and inserted into the gC locus (T. P. Margolis, F. Sedarati, A. T. Dobson, L. T. Feldman, and J. G. Stevens, Virology 189:150-160, 1992). The second virus (KOS-15) was constructed in identical fashion, using plasmid pWAG-15, which is not downregulated by ICP4. Vero cells productively infected with KOS-15 produce 10-fold more β- galactosidase than do those infected with KOS-1. In murine dorsal root ganglia acutely infected with KOS-1, only 1.2{\%} of dorsal root ganglion neurons that expressed viral antigen also expressed β-galactosidase. In contrast, in KOS-15-infected mice, β-galactosidase was detected in 18{\%} of viral antigen-positive neurons. Similar findings were observed in trigeminal ganglia acutely infected with KOS-1 and KOS-15. Thus, the region encompassing the LAT transcription start site appears to play an important role in repression of the LAT promoter activity not only in vitro but also in acutely infected neurons in vivo. These results suggest that during productive infection with HSV-1, LAT expression is tightly regulated.",
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