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Temporal Regulation of Distinct Internal Ribosome Entry Sites of the Dicistroviridae Cricket Paralysis Virus

Internal ribosome entry is a key mechanism for viral protein synthesis in a subset of RNA viruses. Cricket paralysis virus (CrPV), a member of Dicistroviridae, has a positive-sense single strand RNA genome that contains two internal ribosome entry sites (IRES), a 5′untranslated region (5′UTR) and in...

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Autores principales: Khong, Anthony, Bonderoff, Jennifer M., Spriggs, Ruth V., Tammpere, Erik, Kerr, Craig H., Jackson, Thomas J., Willis, Anne E., Jan, Eric
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4728584/
https://www.ncbi.nlm.nih.gov/pubmed/26797630
http://dx.doi.org/10.3390/v8010025
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author Khong, Anthony
Bonderoff, Jennifer M.
Spriggs, Ruth V.
Tammpere, Erik
Kerr, Craig H.
Jackson, Thomas J.
Willis, Anne E.
Jan, Eric
author_facet Khong, Anthony
Bonderoff, Jennifer M.
Spriggs, Ruth V.
Tammpere, Erik
Kerr, Craig H.
Jackson, Thomas J.
Willis, Anne E.
Jan, Eric
author_sort Khong, Anthony
collection PubMed
description Internal ribosome entry is a key mechanism for viral protein synthesis in a subset of RNA viruses. Cricket paralysis virus (CrPV), a member of Dicistroviridae, has a positive-sense single strand RNA genome that contains two internal ribosome entry sites (IRES), a 5′untranslated region (5′UTR) and intergenic region (IGR) IRES, that direct translation of open reading frames (ORF) encoding the viral non-structural and structural proteins, respectively. The regulation of and the significance of the CrPV IRESs during infection are not fully understood. In this study, using a series of biochemical assays including radioactive-pulse labelling, reporter RNA assays and ribosome profiling, we demonstrate that while 5′UTR IRES translational activity is constant throughout infection, IGR IRES translation is delayed and then stimulated two to three hours post infection. The delay in IGR IRES translation is not affected by inhibiting global translation prematurely via treatment with Pateamine A. Using a CrPV replicon that uncouples viral translation and replication, we show that the increase in IGR IRES translation is dependent on expression of non-structural proteins and is greatly stimulated when replication is active. Temporal regulation by distinct IRESs within the CrPV genome is an effective viral strategy to ensure optimal timing and expression of viral proteins to facilitate infection.
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spelling pubmed-47285842016-02-08 Temporal Regulation of Distinct Internal Ribosome Entry Sites of the Dicistroviridae Cricket Paralysis Virus Khong, Anthony Bonderoff, Jennifer M. Spriggs, Ruth V. Tammpere, Erik Kerr, Craig H. Jackson, Thomas J. Willis, Anne E. Jan, Eric Viruses Article Internal ribosome entry is a key mechanism for viral protein synthesis in a subset of RNA viruses. Cricket paralysis virus (CrPV), a member of Dicistroviridae, has a positive-sense single strand RNA genome that contains two internal ribosome entry sites (IRES), a 5′untranslated region (5′UTR) and intergenic region (IGR) IRES, that direct translation of open reading frames (ORF) encoding the viral non-structural and structural proteins, respectively. The regulation of and the significance of the CrPV IRESs during infection are not fully understood. In this study, using a series of biochemical assays including radioactive-pulse labelling, reporter RNA assays and ribosome profiling, we demonstrate that while 5′UTR IRES translational activity is constant throughout infection, IGR IRES translation is delayed and then stimulated two to three hours post infection. The delay in IGR IRES translation is not affected by inhibiting global translation prematurely via treatment with Pateamine A. Using a CrPV replicon that uncouples viral translation and replication, we show that the increase in IGR IRES translation is dependent on expression of non-structural proteins and is greatly stimulated when replication is active. Temporal regulation by distinct IRESs within the CrPV genome is an effective viral strategy to ensure optimal timing and expression of viral proteins to facilitate infection. MDPI 2016-01-19 /pmc/articles/PMC4728584/ /pubmed/26797630 http://dx.doi.org/10.3390/v8010025 Text en © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons by Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Khong, Anthony
Bonderoff, Jennifer M.
Spriggs, Ruth V.
Tammpere, Erik
Kerr, Craig H.
Jackson, Thomas J.
Willis, Anne E.
Jan, Eric
Temporal Regulation of Distinct Internal Ribosome Entry Sites of the Dicistroviridae Cricket Paralysis Virus
title Temporal Regulation of Distinct Internal Ribosome Entry Sites of the Dicistroviridae Cricket Paralysis Virus
title_full Temporal Regulation of Distinct Internal Ribosome Entry Sites of the Dicistroviridae Cricket Paralysis Virus
title_fullStr Temporal Regulation of Distinct Internal Ribosome Entry Sites of the Dicistroviridae Cricket Paralysis Virus
title_full_unstemmed Temporal Regulation of Distinct Internal Ribosome Entry Sites of the Dicistroviridae Cricket Paralysis Virus
title_short Temporal Regulation of Distinct Internal Ribosome Entry Sites of the Dicistroviridae Cricket Paralysis Virus
title_sort temporal regulation of distinct internal ribosome entry sites of the dicistroviridae cricket paralysis virus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4728584/
https://www.ncbi.nlm.nih.gov/pubmed/26797630
http://dx.doi.org/10.3390/v8010025
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