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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...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Public Library of Science
2008
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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 |
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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 |
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