Hepatitis C Virus Translation Preferentially Depends on Active RNA Replication

Hepatitis C virus (HCV) RNA initiates its replication on a detergent-resistant membrane structure derived from the endoplasmic reticulum (ER) in the HCV replicon cells. By performing a pulse-chase study of BrU-labeled HCV RNA, we found that the newly-synthesized HCV RNA traveled along the anterograd...

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Main Authors:	Liu, Helene Minyi, Aizaki, Hideki, Machida, Keigo, Ou, J.-H. James, Lai, Michael M. C.
Format:	Online
Language:	English
Published:	Public Library of Science 2012
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3427374/

id	pubmed-3427374
recordtype	oai_dc
spelling	pubmed-34273742012-08-30 Hepatitis C Virus Translation Preferentially Depends on Active RNA Replication Liu, Helene Minyi Aizaki, Hideki Machida, Keigo Ou, J.-H. James Lai, Michael M. C. Research Article Hepatitis C virus (HCV) RNA initiates its replication on a detergent-resistant membrane structure derived from the endoplasmic reticulum (ER) in the HCV replicon cells. By performing a pulse-chase study of BrU-labeled HCV RNA, we found that the newly-synthesized HCV RNA traveled along the anterograde-membrane traffic and moved away from the ER. Presumably, the RNA moved to the site of translation or virion assembly in the later steps of viral life cycle. In this study, we further addressed how HCV RNA translation was regulated by HCV RNA trafficking. When the movement of HCV RNA from the site of RNA synthesis to the Golgi complex was blocked by nocodazole, an inhibitor of ER-Golgi transport, HCV protein translation was surprisingly enhanced, suggesting that the translation of viral proteins occurred near the site of RNA synthesis. We also found that the translation of HCV proteins was dependent on active RNA synthesis: inhibition of viral RNA synthesis by an NS5B inhibitor resulted in decreased HCV viral protein synthesis even when the total amount of intracellular HCV RNA remained unchanged. Furthermore, the translation activity of the replication-defective HCV replicons or viral RNA with an NS5B mutation was greatly reduced as compared to that of the corresponding wildtype RNA. By performing live cell labeling of newly synthesized HCV RNA and proteins, we further showed that the newly synthesized HCV proteins colocalized with the newly synthesized viral RNA, suggesting that HCV RNA replication and protein translation take place at or near the same site. Our findings together indicate that the translation of HCV RNA is coupled to RNA replication and that the both processes may occur at the same subcellular membrane compartments, which we term the replicasome. Public Library of Science 2012-08-24 /pmc/articles/PMC3427374/ /pubmed/22937067 http://dx.doi.org/10.1371/journal.pone.0043600 Text en © 2012 Liu et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Liu, Helene Minyi Aizaki, Hideki Machida, Keigo Ou, J.-H. James Lai, Michael M. C.
spellingShingle	Liu, Helene Minyi Aizaki, Hideki Machida, Keigo Ou, J.-H. James Lai, Michael M. C. Hepatitis C Virus Translation Preferentially Depends on Active RNA Replication
author_facet	Liu, Helene Minyi Aizaki, Hideki Machida, Keigo Ou, J.-H. James Lai, Michael M. C.
author_sort	Liu, Helene Minyi
title	Hepatitis C Virus Translation Preferentially Depends on Active RNA Replication
title_short	Hepatitis C Virus Translation Preferentially Depends on Active RNA Replication
title_full	Hepatitis C Virus Translation Preferentially Depends on Active RNA Replication
title_fullStr	Hepatitis C Virus Translation Preferentially Depends on Active RNA Replication
title_full_unstemmed	Hepatitis C Virus Translation Preferentially Depends on Active RNA Replication
title_sort	hepatitis c virus translation preferentially depends on active rna replication
description	Hepatitis C virus (HCV) RNA initiates its replication on a detergent-resistant membrane structure derived from the endoplasmic reticulum (ER) in the HCV replicon cells. By performing a pulse-chase study of BrU-labeled HCV RNA, we found that the newly-synthesized HCV RNA traveled along the anterograde-membrane traffic and moved away from the ER. Presumably, the RNA moved to the site of translation or virion assembly in the later steps of viral life cycle. In this study, we further addressed how HCV RNA translation was regulated by HCV RNA trafficking. When the movement of HCV RNA from the site of RNA synthesis to the Golgi complex was blocked by nocodazole, an inhibitor of ER-Golgi transport, HCV protein translation was surprisingly enhanced, suggesting that the translation of viral proteins occurred near the site of RNA synthesis. We also found that the translation of HCV proteins was dependent on active RNA synthesis: inhibition of viral RNA synthesis by an NS5B inhibitor resulted in decreased HCV viral protein synthesis even when the total amount of intracellular HCV RNA remained unchanged. Furthermore, the translation activity of the replication-defective HCV replicons or viral RNA with an NS5B mutation was greatly reduced as compared to that of the corresponding wildtype RNA. By performing live cell labeling of newly synthesized HCV RNA and proteins, we further showed that the newly synthesized HCV proteins colocalized with the newly synthesized viral RNA, suggesting that HCV RNA replication and protein translation take place at or near the same site. Our findings together indicate that the translation of HCV RNA is coupled to RNA replication and that the both processes may occur at the same subcellular membrane compartments, which we term the replicasome.
publisher	Public Library of Science
publishDate	2012
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3427374/
_version_	1611552167958675456

Hepatitis C Virus Translation Preferentially Depends on Active RNA Replication

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