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Title: Inhibition of iridovirus protein synthesis and virus replication by antisense morpholino oligonucleotides targeted to the major capsid protein, the 18 kDa immediate-early protein, and a viral homolog of RNA polymerase II

Abstract

Frog virus 3 (FV3) is a large DNA virus that encodes {approx} 100 proteins. Although the general features of FV3 replication are known, the specific roles that most viral proteins play in the virus life cycle have not yet been elucidated. To address the question of viral gene function, antisense morpholino oligonucleotides (asMOs) were used to transiently knock-down expression of specific viral genes and thus infer their role in virus replication. We designed asMOs directed against the major capsid protein (MCP), an 18 kDa immediate-early protein (18K) that was thought to be a viral regulatory protein, and the viral homologue of the largest subunit of RNA polymerase II (vPol-II{alpha}). All three asMOs successfully inhibited translation of the targeted protein, and two of the three asMOs resulted in marked phenotypic changes. Knock-down of the MCP resulted in a marked reduction in viral titer without a corresponding drop in the synthesis of other late viral proteins. Transmission electron microscopy (TEM) showed that in cells treated with the anti-MCP MO assembly sites were devoid of viral particles and contained numerous aberrant structures. In contrast, inhibition of 18K synthesis did not block virion formation, suggesting that the 18K protein was not essential for replicationmore » of FV3 in fathead minnow (FHM) cells. Finally, consistent with the view that late viral gene expression is catalyzed by a virus-encoded or virus-modified Pol-II-like protein, knock-down of vPol-II{alpha} triggered a global decline in late gene expression and virus yields without affecting the synthesis of early viral genes. Collectively, these results demonstrate the utility of using asMOs to elucidate the function of FV3 proteins.« less

Authors:
 [1];  [1];  [1];  [1];  [2];  [2];  [3];  [1]
  1. Department of Microbiology, University of Mississippi Medical Center, Jackson, MS 39216 (United States)
  2. Department of Anatomy, University of Mississippi Medical Center, Jackson, MS 39216 (United States)
  3. Department of Preventive Medicine, University of Mississippi Medical Center, Jackson, MS 39216 (United States)
Publication Date:
OSTI Identifier:
20975215
Resource Type:
Journal Article
Journal Name:
Virology
Additional Journal Information:
Journal Volume: 358; Journal Issue: 2; Other Information: DOI: 10.1016/j.virol.2006.07.009; PII: S0042-6822(06)00449-1; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0042-6822
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; BIOSYNTHESIS; FATHEAD MINNOW; GENES; INHIBITION; LIFE CYCLE; OLIGONUCLEOTIDES; RNA POLYMERASES; TRANSMISSION ELECTRON MICROSCOPY; VIRUSES

Citation Formats

Sample, Robert, Bryan, Locke, Long, Scott, Majji, Sai, Hoskins, Glenn, Sinning, Allan, Olivier, Jake, and Chinchar, V Gregory. Inhibition of iridovirus protein synthesis and virus replication by antisense morpholino oligonucleotides targeted to the major capsid protein, the 18 kDa immediate-early protein, and a viral homolog of RNA polymerase II. United States: N. p., 2007. Web. doi:10.1016/j.virol.2006.07.009.
Sample, Robert, Bryan, Locke, Long, Scott, Majji, Sai, Hoskins, Glenn, Sinning, Allan, Olivier, Jake, & Chinchar, V Gregory. Inhibition of iridovirus protein synthesis and virus replication by antisense morpholino oligonucleotides targeted to the major capsid protein, the 18 kDa immediate-early protein, and a viral homolog of RNA polymerase II. United States. https://doi.org/10.1016/j.virol.2006.07.009
Sample, Robert, Bryan, Locke, Long, Scott, Majji, Sai, Hoskins, Glenn, Sinning, Allan, Olivier, Jake, and Chinchar, V Gregory. 2007. "Inhibition of iridovirus protein synthesis and virus replication by antisense morpholino oligonucleotides targeted to the major capsid protein, the 18 kDa immediate-early protein, and a viral homolog of RNA polymerase II". United States. https://doi.org/10.1016/j.virol.2006.07.009.
@article{osti_20975215,
title = {Inhibition of iridovirus protein synthesis and virus replication by antisense morpholino oligonucleotides targeted to the major capsid protein, the 18 kDa immediate-early protein, and a viral homolog of RNA polymerase II},
author = {Sample, Robert and Bryan, Locke and Long, Scott and Majji, Sai and Hoskins, Glenn and Sinning, Allan and Olivier, Jake and Chinchar, V Gregory},
abstractNote = {Frog virus 3 (FV3) is a large DNA virus that encodes {approx} 100 proteins. Although the general features of FV3 replication are known, the specific roles that most viral proteins play in the virus life cycle have not yet been elucidated. To address the question of viral gene function, antisense morpholino oligonucleotides (asMOs) were used to transiently knock-down expression of specific viral genes and thus infer their role in virus replication. We designed asMOs directed against the major capsid protein (MCP), an 18 kDa immediate-early protein (18K) that was thought to be a viral regulatory protein, and the viral homologue of the largest subunit of RNA polymerase II (vPol-II{alpha}). All three asMOs successfully inhibited translation of the targeted protein, and two of the three asMOs resulted in marked phenotypic changes. Knock-down of the MCP resulted in a marked reduction in viral titer without a corresponding drop in the synthesis of other late viral proteins. Transmission electron microscopy (TEM) showed that in cells treated with the anti-MCP MO assembly sites were devoid of viral particles and contained numerous aberrant structures. In contrast, inhibition of 18K synthesis did not block virion formation, suggesting that the 18K protein was not essential for replication of FV3 in fathead minnow (FHM) cells. Finally, consistent with the view that late viral gene expression is catalyzed by a virus-encoded or virus-modified Pol-II-like protein, knock-down of vPol-II{alpha} triggered a global decline in late gene expression and virus yields without affecting the synthesis of early viral genes. Collectively, these results demonstrate the utility of using asMOs to elucidate the function of FV3 proteins.},
doi = {10.1016/j.virol.2006.07.009},
url = {https://www.osti.gov/biblio/20975215}, journal = {Virology},
issn = {0042-6822},
number = 2,
volume = 358,
place = {United States},
year = {Tue Feb 20 00:00:00 EST 2007},
month = {Tue Feb 20 00:00:00 EST 2007}
}