The Heteroaryldihydropyrimidine Bay 38-7690 Induces Hepatitis B Virus Core Protein Aggregates Associated with Promyelocytic Leukemia Nuclear Bodies in Infected Cells

ABSTRACT Heteroaryldihydropyrimidines (HAPs) are compounds that inhibit hepatitis B virus (HBV) replication by modulating viral capsid assembly. While their biophysical effects on capsid assembly in vitro have been previously studied, the effect of HAP treatment on capsid protein (Cp) in individual...

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Autores principales:	Andrew D. Huber, Jennifer J. Wolf, Dandan Liu, Anna T. Gres, Jing Tang, Kelsey N. Boschert, Maritza N. Puray-Chavez, Dallas L. Pineda, Thomas G. Laughlin, Emily M. Coonrod, Qiongying Yang, Juan Ji, Karen A. Kirby, Zhengqiang Wang, Stefan G. Sarafianos
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	antiviral agents capsid hepatitis B virus mechanisms of action virology virus-host interactions Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/d21fb67e4b2b429c9a5549748928bef6
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spelling	oai:doaj.org-article:d21fb67e4b2b429c9a5549748928bef62021-11-15T15:22:13ZThe Heteroaryldihydropyrimidine Bay 38-7690 Induces Hepatitis B Virus Core Protein Aggregates Associated with Promyelocytic Leukemia Nuclear Bodies in Infected Cells10.1128/mSphereDirect.00131-182379-5042https://doaj.org/article/d21fb67e4b2b429c9a5549748928bef62018-04-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphereDirect.00131-18https://doaj.org/toc/2379-5042ABSTRACT Heteroaryldihydropyrimidines (HAPs) are compounds that inhibit hepatitis B virus (HBV) replication by modulating viral capsid assembly. While their biophysical effects on capsid assembly in vitro have been previously studied, the effect of HAP treatment on capsid protein (Cp) in individual HBV-infected cells remains unknown. We report here that the HAP Bay 38-7690 promotes aggregation of recombinant Cp in vitro and causes a time- and dose-dependent decrease of Cp in infected cells, consistent with previously studied HAPs. Interestingly, immunofluorescence analysis showed Cp aggregating in nuclear foci of Bay 38-7690-treated infected cells in a time- and dose-dependent manner. We found these foci to be associated with promyelocytic leukemia (PML) nuclear bodies (NBs), which are structures that affect many cellular functions, including DNA damage response, transcription, apoptosis, and antiviral responses. Cp aggregation is not an artifact of the cell system used, as it is observed in HBV-expressing HepAD38 cells, in HepG2 cells transfected with an HBV-expressing plasmid, and in HepG2-NTCP cells infected with HBV. Use of a Cp overexpression vector without HBV sequences shows that aggregation is independent of viral replication, and use of an HBV-expressing plasmid harboring a HAP resistance mutation in Cp abrogated the aggregation, demonstrating that the effect is due to direct compound-Cp interactions. These studies provide novel insight into the effects of HAP-based treatment at a single-cell level. IMPORTANCE Despite the availability of effective vaccines and treatments, HBV remains a significant global health concern, with more than 240 million individuals chronically infected. Current treatments are highly effective at controlling viral replication and disease progression but rarely cure infections. Therefore, much emphasis is being placed on finding therapeutics with new drug targets, such as viral gene expression, covalently closed circular DNA formation and stability, capsid formation, and host immune modulators, with the ultimate goal of an HBV cure. Understanding the mechanisms by which novel antiviral agents act will be imperative for the development of curative HBV therapies.Andrew D. HuberJennifer J. WolfDandan LiuAnna T. GresJing TangKelsey N. BoschertMaritza N. Puray-ChavezDallas L. PinedaThomas G. LaughlinEmily M. CoonrodQiongying YangJuan JiKaren A. KirbyZhengqiang WangStefan G. SarafianosAmerican Society for Microbiologyarticleantiviral agentscapsidhepatitis B virusmechanisms of actionvirologyvirus-host interactionsMicrobiologyQR1-502ENmSphere, Vol 3, Iss 2 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	antiviral agents capsid hepatitis B virus mechanisms of action virology virus-host interactions Microbiology QR1-502
spellingShingle	antiviral agents capsid hepatitis B virus mechanisms of action virology virus-host interactions Microbiology QR1-502 Andrew D. Huber Jennifer J. Wolf Dandan Liu Anna T. Gres Jing Tang Kelsey N. Boschert Maritza N. Puray-Chavez Dallas L. Pineda Thomas G. Laughlin Emily M. Coonrod Qiongying Yang Juan Ji Karen A. Kirby Zhengqiang Wang Stefan G. Sarafianos The Heteroaryldihydropyrimidine Bay 38-7690 Induces Hepatitis B Virus Core Protein Aggregates Associated with Promyelocytic Leukemia Nuclear Bodies in Infected Cells
description	ABSTRACT Heteroaryldihydropyrimidines (HAPs) are compounds that inhibit hepatitis B virus (HBV) replication by modulating viral capsid assembly. While their biophysical effects on capsid assembly in vitro have been previously studied, the effect of HAP treatment on capsid protein (Cp) in individual HBV-infected cells remains unknown. We report here that the HAP Bay 38-7690 promotes aggregation of recombinant Cp in vitro and causes a time- and dose-dependent decrease of Cp in infected cells, consistent with previously studied HAPs. Interestingly, immunofluorescence analysis showed Cp aggregating in nuclear foci of Bay 38-7690-treated infected cells in a time- and dose-dependent manner. We found these foci to be associated with promyelocytic leukemia (PML) nuclear bodies (NBs), which are structures that affect many cellular functions, including DNA damage response, transcription, apoptosis, and antiviral responses. Cp aggregation is not an artifact of the cell system used, as it is observed in HBV-expressing HepAD38 cells, in HepG2 cells transfected with an HBV-expressing plasmid, and in HepG2-NTCP cells infected with HBV. Use of a Cp overexpression vector without HBV sequences shows that aggregation is independent of viral replication, and use of an HBV-expressing plasmid harboring a HAP resistance mutation in Cp abrogated the aggregation, demonstrating that the effect is due to direct compound-Cp interactions. These studies provide novel insight into the effects of HAP-based treatment at a single-cell level. IMPORTANCE Despite the availability of effective vaccines and treatments, HBV remains a significant global health concern, with more than 240 million individuals chronically infected. Current treatments are highly effective at controlling viral replication and disease progression but rarely cure infections. Therefore, much emphasis is being placed on finding therapeutics with new drug targets, such as viral gene expression, covalently closed circular DNA formation and stability, capsid formation, and host immune modulators, with the ultimate goal of an HBV cure. Understanding the mechanisms by which novel antiviral agents act will be imperative for the development of curative HBV therapies.
format	article
author	Andrew D. Huber Jennifer J. Wolf Dandan Liu Anna T. Gres Jing Tang Kelsey N. Boschert Maritza N. Puray-Chavez Dallas L. Pineda Thomas G. Laughlin Emily M. Coonrod Qiongying Yang Juan Ji Karen A. Kirby Zhengqiang Wang Stefan G. Sarafianos
author_facet	Andrew D. Huber Jennifer J. Wolf Dandan Liu Anna T. Gres Jing Tang Kelsey N. Boschert Maritza N. Puray-Chavez Dallas L. Pineda Thomas G. Laughlin Emily M. Coonrod Qiongying Yang Juan Ji Karen A. Kirby Zhengqiang Wang Stefan G. Sarafianos
author_sort	Andrew D. Huber
title	The Heteroaryldihydropyrimidine Bay 38-7690 Induces Hepatitis B Virus Core Protein Aggregates Associated with Promyelocytic Leukemia Nuclear Bodies in Infected Cells
title_short	The Heteroaryldihydropyrimidine Bay 38-7690 Induces Hepatitis B Virus Core Protein Aggregates Associated with Promyelocytic Leukemia Nuclear Bodies in Infected Cells
title_full	The Heteroaryldihydropyrimidine Bay 38-7690 Induces Hepatitis B Virus Core Protein Aggregates Associated with Promyelocytic Leukemia Nuclear Bodies in Infected Cells
title_fullStr	The Heteroaryldihydropyrimidine Bay 38-7690 Induces Hepatitis B Virus Core Protein Aggregates Associated with Promyelocytic Leukemia Nuclear Bodies in Infected Cells
title_full_unstemmed	The Heteroaryldihydropyrimidine Bay 38-7690 Induces Hepatitis B Virus Core Protein Aggregates Associated with Promyelocytic Leukemia Nuclear Bodies in Infected Cells
title_sort	heteroaryldihydropyrimidine bay 38-7690 induces hepatitis b virus core protein aggregates associated with promyelocytic leukemia nuclear bodies in infected cells
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/d21fb67e4b2b429c9a5549748928bef6
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The Heteroaryldihydropyrimidine Bay 38-7690 Induces Hepatitis B Virus Core Protein Aggregates Associated with Promyelocytic Leukemia Nuclear Bodies in Infected Cells

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