Cargando…

Ubiquitination Is Required for Effective Replication of Coxsackievirus B3

BACKGROUND: Protein ubiquitination and/or degradation by the ubiquitin/proteasome system (UPS) have been recognized as critical mechanisms in the regulation of numerous essential cellular functions. The importance of the UPS in viral pathogenesis has become increasingly apparent. Using murine cardio...

Descripción completa

Detalles Bibliográficos
Autores principales: Si, Xiaoning, Gao, Guang, Wong, Jerry, Wang, Yahong, Zhang, Jingchun, Luo, Honglin
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2440516/
https://www.ncbi.nlm.nih.gov/pubmed/18612413
http://dx.doi.org/10.1371/journal.pone.0002585
_version_ 1782156551082999808
author Si, Xiaoning
Gao, Guang
Wong, Jerry
Wang, Yahong
Zhang, Jingchun
Luo, Honglin
author_facet Si, Xiaoning
Gao, Guang
Wong, Jerry
Wang, Yahong
Zhang, Jingchun
Luo, Honglin
author_sort Si, Xiaoning
collection PubMed
description BACKGROUND: Protein ubiquitination and/or degradation by the ubiquitin/proteasome system (UPS) have been recognized as critical mechanisms in the regulation of numerous essential cellular functions. The importance of the UPS in viral pathogenesis has become increasingly apparent. Using murine cardiomyocytes, we have previously demonstrated that the UPS plays a key role in the replication of coxsackievirus B3 (CVB3), an important human pathogen associated with various diseases. To further elucidate the underlying mechanisms, we examined the interplay between the UPS and CVB3, focusing on the role of ubiquitination in viral lifecycle. METHODOLOGY/PRINCIPAL FINDINGS: As assessed by in situ hybridization, Western blot, and plaque assay, we showed that proteasome inhibition decreased CVB3 RNA replication, protein synthesis, and viral titers in HeLa cells. There were no apparent changes in 20S proteasome activities following CVB3 infection. However, we found viral infection led to an accumulation of protein-ubiquitin conjugates, accompanied by a decreased protein expression of free ubiquitin, implicating an important role of ubiquitination in the UPS-mediated viral replication. Using small-interfering RNA, we demonstrated that gene-silencing of ubiquitin significantly reduced viral titers, possibly through downregulation of protein ubiquitination and subsequent alteration of protein function and/or degradation. Inhibition of deubiquitinating enzymes apparently enhances the inhibitory effects of proteasome inhibitors on CVB3 replication. Finally, by immunoprecipitation, we showed that coxsackieviral polymerase 3D was post-translationally modified by ubiquitination and such modification might be a prerequisite for its function in transcriptional regulation of viral genome. CONCLUSION: Coxsackievirus infection promotes protein ubiquitination, contributing to effective viral replication, probably through ubiquitin modification of viral polymerase.
format Text
id pubmed-2440516
institution National Center for Biotechnology Information
language English
publishDate 2008
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-24405162008-07-09 Ubiquitination Is Required for Effective Replication of Coxsackievirus B3 Si, Xiaoning Gao, Guang Wong, Jerry Wang, Yahong Zhang, Jingchun Luo, Honglin PLoS One Research Article BACKGROUND: Protein ubiquitination and/or degradation by the ubiquitin/proteasome system (UPS) have been recognized as critical mechanisms in the regulation of numerous essential cellular functions. The importance of the UPS in viral pathogenesis has become increasingly apparent. Using murine cardiomyocytes, we have previously demonstrated that the UPS plays a key role in the replication of coxsackievirus B3 (CVB3), an important human pathogen associated with various diseases. To further elucidate the underlying mechanisms, we examined the interplay between the UPS and CVB3, focusing on the role of ubiquitination in viral lifecycle. METHODOLOGY/PRINCIPAL FINDINGS: As assessed by in situ hybridization, Western blot, and plaque assay, we showed that proteasome inhibition decreased CVB3 RNA replication, protein synthesis, and viral titers in HeLa cells. There were no apparent changes in 20S proteasome activities following CVB3 infection. However, we found viral infection led to an accumulation of protein-ubiquitin conjugates, accompanied by a decreased protein expression of free ubiquitin, implicating an important role of ubiquitination in the UPS-mediated viral replication. Using small-interfering RNA, we demonstrated that gene-silencing of ubiquitin significantly reduced viral titers, possibly through downregulation of protein ubiquitination and subsequent alteration of protein function and/or degradation. Inhibition of deubiquitinating enzymes apparently enhances the inhibitory effects of proteasome inhibitors on CVB3 replication. Finally, by immunoprecipitation, we showed that coxsackieviral polymerase 3D was post-translationally modified by ubiquitination and such modification might be a prerequisite for its function in transcriptional regulation of viral genome. CONCLUSION: Coxsackievirus infection promotes protein ubiquitination, contributing to effective viral replication, probably through ubiquitin modification of viral polymerase. Public Library of Science 2008-07-09 /pmc/articles/PMC2440516/ /pubmed/18612413 http://dx.doi.org/10.1371/journal.pone.0002585 Text en Si 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.
spellingShingle Research Article
Si, Xiaoning
Gao, Guang
Wong, Jerry
Wang, Yahong
Zhang, Jingchun
Luo, Honglin
Ubiquitination Is Required for Effective Replication of Coxsackievirus B3
title Ubiquitination Is Required for Effective Replication of Coxsackievirus B3
title_full Ubiquitination Is Required for Effective Replication of Coxsackievirus B3
title_fullStr Ubiquitination Is Required for Effective Replication of Coxsackievirus B3
title_full_unstemmed Ubiquitination Is Required for Effective Replication of Coxsackievirus B3
title_short Ubiquitination Is Required for Effective Replication of Coxsackievirus B3
title_sort ubiquitination is required for effective replication of coxsackievirus b3
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2440516/
https://www.ncbi.nlm.nih.gov/pubmed/18612413
http://dx.doi.org/10.1371/journal.pone.0002585
work_keys_str_mv AT sixiaoning ubiquitinationisrequiredforeffectivereplicationofcoxsackievirusb3
AT gaoguang ubiquitinationisrequiredforeffectivereplicationofcoxsackievirusb3
AT wongjerry ubiquitinationisrequiredforeffectivereplicationofcoxsackievirusb3
AT wangyahong ubiquitinationisrequiredforeffectivereplicationofcoxsackievirusb3
AT zhangjingchun ubiquitinationisrequiredforeffectivereplicationofcoxsackievirusb3
AT luohonglin ubiquitinationisrequiredforeffectivereplicationofcoxsackievirusb3