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Title: Characterization of a spliced exon product of herpes simplex type-1 latency-associated transcript in productively infected cells

Abstract

The latency-associated transcripts (LATs) of herpes simplex virus type-1 (HSV-1) are the only viral RNAs accumulating during latent infections in the sensory ganglia of the peripheral nervous system. The major form of LAT that accumulates in latently infected neurons is a 2 kb intron, spliced from a much less abundant 8.3 primary transcript. The spliced exon mRNA has been hard to detect. However, in this study, we have examined the spliced exon RNA in productively infected cells using ribonuclease protection (RPA), and quantitative RT-PCR (q-PCR) assays. We were able to detect the LAT exon RNA in productively infected SY5Y cells (a human neuronal cell line). The level of the LAT exon RNA was found to be approximately 5% that of the 2 kb intron RNA and thus is likely to be relatively unstable. Quantitative RT-PCR (q-PCR) assays were used to examine the LAT exon RNA and its properties. They confirmed that the LAT exon mRNA is present at a very low level in productively infected cells, compared to the levels of other viral transcripts. Furthermore, experiments showed that the LAT exon mRNA is expressed as a true late gene, and appears to be polyadenylated. In SY5Y cells, in contrast tomore » most late viral transcripts, the LAT exon RNA was found to be mainly nuclear localized during the late stage of a productive infection. Interestingly, more LAT exon RNA was found in the cytoplasm in differentiated compared to undifferentiated SY5Y cells, suggesting the nucleocytoplasmic distribution of the LAT exon RNA and its related function may be influenced by the differentiation state of cells.« less

Authors:
 [1];  [1];  [1];  [2];  [3];  [4]
  1. Department of Microbiology, University of Pennsylvania Medical School, 315 Johnson Pavilion, Philadelphia, PA 19104 (United States)
  2. Department of Genetics, University of Pennsylvania School of Medicine, Philadelphia, PA 19104 (United States)
  3. Department of Microbiology and Molecular Genetics, College of Medicine, University of California, Irvine, CA 92697-4025 (United States)
  4. Department of Microbiology, University of Pennsylvania Medical School, 315 Johnson Pavilion, Philadelphia, PA 19104 (United States). E-mail: nfraser@mail.med.upenn.edu
Publication Date:
OSTI Identifier:
20850579
Resource Type:
Journal Article
Resource Relation:
Journal Name: Virology; Journal Volume: 356; Journal Issue: 1-2; Other Information: DOI: 10.1016/j.virol.2006.07.033; PII: S0042-6822(06)00510-1; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CYTOPLASM; HERPES SIMPLEX; NERVE CELLS; NERVOUS SYSTEM; POLYMERASE CHAIN REACTION; RNA; RNA-ASE; VIRUSES

Citation Formats

Kang, Wen, Mukerjee, Ruma, Gartner, Jared J., Hatzigeorgiou, Artemis G., Sandri-Goldin, Rozanne M., and Fraser, Nigel W.. Characterization of a spliced exon product of herpes simplex type-1 latency-associated transcript in productively infected cells. United States: N. p., 2006. Web. doi:10.1016/j.virol.2006.07.033.
Kang, Wen, Mukerjee, Ruma, Gartner, Jared J., Hatzigeorgiou, Artemis G., Sandri-Goldin, Rozanne M., & Fraser, Nigel W.. Characterization of a spliced exon product of herpes simplex type-1 latency-associated transcript in productively infected cells. United States. doi:10.1016/j.virol.2006.07.033.
Kang, Wen, Mukerjee, Ruma, Gartner, Jared J., Hatzigeorgiou, Artemis G., Sandri-Goldin, Rozanne M., and Fraser, Nigel W.. Wed . "Characterization of a spliced exon product of herpes simplex type-1 latency-associated transcript in productively infected cells". United States. doi:10.1016/j.virol.2006.07.033.
@article{osti_20850579,
title = {Characterization of a spliced exon product of herpes simplex type-1 latency-associated transcript in productively infected cells},
author = {Kang, Wen and Mukerjee, Ruma and Gartner, Jared J. and Hatzigeorgiou, Artemis G. and Sandri-Goldin, Rozanne M. and Fraser, Nigel W.},
abstractNote = {The latency-associated transcripts (LATs) of herpes simplex virus type-1 (HSV-1) are the only viral RNAs accumulating during latent infections in the sensory ganglia of the peripheral nervous system. The major form of LAT that accumulates in latently infected neurons is a 2 kb intron, spliced from a much less abundant 8.3 primary transcript. The spliced exon mRNA has been hard to detect. However, in this study, we have examined the spliced exon RNA in productively infected cells using ribonuclease protection (RPA), and quantitative RT-PCR (q-PCR) assays. We were able to detect the LAT exon RNA in productively infected SY5Y cells (a human neuronal cell line). The level of the LAT exon RNA was found to be approximately 5% that of the 2 kb intron RNA and thus is likely to be relatively unstable. Quantitative RT-PCR (q-PCR) assays were used to examine the LAT exon RNA and its properties. They confirmed that the LAT exon mRNA is present at a very low level in productively infected cells, compared to the levels of other viral transcripts. Furthermore, experiments showed that the LAT exon mRNA is expressed as a true late gene, and appears to be polyadenylated. In SY5Y cells, in contrast to most late viral transcripts, the LAT exon RNA was found to be mainly nuclear localized during the late stage of a productive infection. Interestingly, more LAT exon RNA was found in the cytoplasm in differentiated compared to undifferentiated SY5Y cells, suggesting the nucleocytoplasmic distribution of the LAT exon RNA and its related function may be influenced by the differentiation state of cells.},
doi = {10.1016/j.virol.2006.07.033},
journal = {Virology},
number = 1-2,
volume = 356,
place = {United States},
year = {Wed Dec 20 00:00:00 EST 2006},
month = {Wed Dec 20 00:00:00 EST 2006}
}
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