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RNase P Ribozymes Inhibit the Replication of Human Cytomegalovirus by Targeting Essential Viral Capsid Proteins

An engineered RNase P-based ribozyme variant, which was generated using the in vitro selection procedure, was used to target the overlapping mRNA region of two proteins essential for human cytomegalovirus (HCMV) replication: capsid assembly protein (AP) and protease (PR). In vitro studies showed tha...

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Autores principales: Yang, Zhu, Reeves, Michael, Ye, Jun, Trang, Phong, Zhu, Li, Sheng, Jingxue, Wang, Yu, Zen, Ke, Wu, Jianguo, Liu, Fenyong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4517104/
https://www.ncbi.nlm.nih.gov/pubmed/26114473
http://dx.doi.org/10.3390/v7072775
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author Yang, Zhu
Reeves, Michael
Ye, Jun
Trang, Phong
Zhu, Li
Sheng, Jingxue
Wang, Yu
Zen, Ke
Wu, Jianguo
Liu, Fenyong
author_facet Yang, Zhu
Reeves, Michael
Ye, Jun
Trang, Phong
Zhu, Li
Sheng, Jingxue
Wang, Yu
Zen, Ke
Wu, Jianguo
Liu, Fenyong
author_sort Yang, Zhu
collection PubMed
description An engineered RNase P-based ribozyme variant, which was generated using the in vitro selection procedure, was used to target the overlapping mRNA region of two proteins essential for human cytomegalovirus (HCMV) replication: capsid assembly protein (AP) and protease (PR). In vitro studies showed that the generated variant, V718-A, cleaved the target AP mRNA sequence efficiently and its activity was about 60-fold higher than that of wild type ribozyme M1-A. Furthermore, we observed a reduction of 98%–99% in AP/PR expression and an inhibition of 50,000 fold in viral growth in cells with V718-A, while a 75% reduction in AP/PR expression and a 500-fold inhibition in viral growth was found in cells with M1-A. Examination of the antiviral effects of the generated ribozyme on the HCMV replication cycle suggested that viral DNA encapsidation was inhibited and as a consequence, viral capsid assembly was blocked when the expression of AP and PR was inhibited by the ribozyme. Thus, our study indicates that the generated ribozyme variant is highly effective in inhibiting HCMV gene expression and blocking viral replication, and suggests that engineered RNase P ribozyme can be potentially developed as a promising gene-targeting agent for anti-HCMV therapy.
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spelling pubmed-45171042015-07-28 RNase P Ribozymes Inhibit the Replication of Human Cytomegalovirus by Targeting Essential Viral Capsid Proteins Yang, Zhu Reeves, Michael Ye, Jun Trang, Phong Zhu, Li Sheng, Jingxue Wang, Yu Zen, Ke Wu, Jianguo Liu, Fenyong Viruses Article An engineered RNase P-based ribozyme variant, which was generated using the in vitro selection procedure, was used to target the overlapping mRNA region of two proteins essential for human cytomegalovirus (HCMV) replication: capsid assembly protein (AP) and protease (PR). In vitro studies showed that the generated variant, V718-A, cleaved the target AP mRNA sequence efficiently and its activity was about 60-fold higher than that of wild type ribozyme M1-A. Furthermore, we observed a reduction of 98%–99% in AP/PR expression and an inhibition of 50,000 fold in viral growth in cells with V718-A, while a 75% reduction in AP/PR expression and a 500-fold inhibition in viral growth was found in cells with M1-A. Examination of the antiviral effects of the generated ribozyme on the HCMV replication cycle suggested that viral DNA encapsidation was inhibited and as a consequence, viral capsid assembly was blocked when the expression of AP and PR was inhibited by the ribozyme. Thus, our study indicates that the generated ribozyme variant is highly effective in inhibiting HCMV gene expression and blocking viral replication, and suggests that engineered RNase P ribozyme can be potentially developed as a promising gene-targeting agent for anti-HCMV therapy. MDPI 2015-06-24 /pmc/articles/PMC4517104/ /pubmed/26114473 http://dx.doi.org/10.3390/v7072775 Text en © 2015 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Yang, Zhu
Reeves, Michael
Ye, Jun
Trang, Phong
Zhu, Li
Sheng, Jingxue
Wang, Yu
Zen, Ke
Wu, Jianguo
Liu, Fenyong
RNase P Ribozymes Inhibit the Replication of Human Cytomegalovirus by Targeting Essential Viral Capsid Proteins
title RNase P Ribozymes Inhibit the Replication of Human Cytomegalovirus by Targeting Essential Viral Capsid Proteins
title_full RNase P Ribozymes Inhibit the Replication of Human Cytomegalovirus by Targeting Essential Viral Capsid Proteins
title_fullStr RNase P Ribozymes Inhibit the Replication of Human Cytomegalovirus by Targeting Essential Viral Capsid Proteins
title_full_unstemmed RNase P Ribozymes Inhibit the Replication of Human Cytomegalovirus by Targeting Essential Viral Capsid Proteins
title_short RNase P Ribozymes Inhibit the Replication of Human Cytomegalovirus by Targeting Essential Viral Capsid Proteins
title_sort rnase p ribozymes inhibit the replication of human cytomegalovirus by targeting essential viral capsid proteins
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4517104/
https://www.ncbi.nlm.nih.gov/pubmed/26114473
http://dx.doi.org/10.3390/v7072775
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