Berberine Chloride is an Alphavirus Inhibitor That Targets Nucleocapsid Assembly

ABSTRACT Alphaviruses are enveloped positive-sense RNA viruses that can cause serious human illnesses such as polyarthritis and encephalitis. Despite their widespread distribution and medical importance, there are no licensed vaccines or antivirals to combat alphavirus infections. Berberine chloride...

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Autores principales:	Judy J. Wan, Rebecca S. Brown, Margaret Kielian
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	berberine RNA packaging alphavirus antiviral inhibitor nucleocapsid assembly Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/f23789612e504d87af66ef62aa096096
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spelling	oai:doaj.org-article:f23789612e504d87af66ef62aa0960962021-11-15T15:56:47ZBerberine Chloride is an Alphavirus Inhibitor That Targets Nucleocapsid Assembly10.1128/mBio.01382-202150-7511https://doaj.org/article/f23789612e504d87af66ef62aa0960962020-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01382-20https://doaj.org/toc/2150-7511ABSTRACT Alphaviruses are enveloped positive-sense RNA viruses that can cause serious human illnesses such as polyarthritis and encephalitis. Despite their widespread distribution and medical importance, there are no licensed vaccines or antivirals to combat alphavirus infections. Berberine chloride (BBC) is a pan-alphavirus inhibitor that was previously identified in a replicon-based small-molecule screen. This work showed that BBC inhibits alphavirus replication but also suggested that BBC might have additional effects later in the viral life cycle. Here, we show that BBC has late effects that target the virus nucleocapsid (NC) core. Infected cells treated with BBC late in infection were unable to form stable cytoplasmic NCs or assembly intermediates, as assayed by gradient sedimentation. In vitro studies with recombinant capsid protein (Cp) and purified genomic RNA (gRNA) showed that BBC perturbs core-like particle formation and potentially traps the assembly process in intermediate states. Particles produced from BBC-treated cells were less infectious, despite efficient particle production and only minor decreases in genome packaging. In addition, BBC treatment of free virus particles strongly decreased alphavirus infectivity. In contrast, the infectivity of the negative-sense RNA virus vesicular stomatitis virus was resistant to BBC treatment of infected cells or free virus. Together, our data indicate that BBC alters alphavirus Cp-gRNA interactions and oligomerization and suggest that this may cause defects in NC assembly and in disassembly during subsequent virus entry. Thus, BBC may be considered a novel alphavirus NC assembly inhibitor. IMPORTANCE The alphavirus chikungunya virus (CHIKV) is an example of an emerging human pathogen with increased and rapid global spread. Although an acute CHIKV infection is rarely fatal, many patients suffer from debilitating chronic arthralgia for years. Antivirals against chikungunya and other alphaviruses have been identified in vitro, but to date none have been shown to be efficacious and have been licensed for human use. Here, we investigated a small molecule, berberine chloride (BBC), and showed that it inhibited infectious virus production by several alphaviruses including CHIKV. BBC acted on a late step in the alphavirus exit pathway, namely the formation of the nucleocapsid containing the infectious viral RNA. Better understanding of nucleocapsid formation and its inhibition by BBC will provide important information on the mechanisms of infectious alphavirus production and may enable their future targeting in antiviral strategies.Judy J. WanRebecca S. BrownMargaret KielianAmerican Society for MicrobiologyarticleberberineRNA packagingalphavirusantiviral inhibitornucleocapsid assemblyMicrobiologyQR1-502ENmBio, Vol 11, Iss 3 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	berberine RNA packaging alphavirus antiviral inhibitor nucleocapsid assembly Microbiology QR1-502
spellingShingle	berberine RNA packaging alphavirus antiviral inhibitor nucleocapsid assembly Microbiology QR1-502 Judy J. Wan Rebecca S. Brown Margaret Kielian Berberine Chloride is an Alphavirus Inhibitor That Targets Nucleocapsid Assembly
description	ABSTRACT Alphaviruses are enveloped positive-sense RNA viruses that can cause serious human illnesses such as polyarthritis and encephalitis. Despite their widespread distribution and medical importance, there are no licensed vaccines or antivirals to combat alphavirus infections. Berberine chloride (BBC) is a pan-alphavirus inhibitor that was previously identified in a replicon-based small-molecule screen. This work showed that BBC inhibits alphavirus replication but also suggested that BBC might have additional effects later in the viral life cycle. Here, we show that BBC has late effects that target the virus nucleocapsid (NC) core. Infected cells treated with BBC late in infection were unable to form stable cytoplasmic NCs or assembly intermediates, as assayed by gradient sedimentation. In vitro studies with recombinant capsid protein (Cp) and purified genomic RNA (gRNA) showed that BBC perturbs core-like particle formation and potentially traps the assembly process in intermediate states. Particles produced from BBC-treated cells were less infectious, despite efficient particle production and only minor decreases in genome packaging. In addition, BBC treatment of free virus particles strongly decreased alphavirus infectivity. In contrast, the infectivity of the negative-sense RNA virus vesicular stomatitis virus was resistant to BBC treatment of infected cells or free virus. Together, our data indicate that BBC alters alphavirus Cp-gRNA interactions and oligomerization and suggest that this may cause defects in NC assembly and in disassembly during subsequent virus entry. Thus, BBC may be considered a novel alphavirus NC assembly inhibitor. IMPORTANCE The alphavirus chikungunya virus (CHIKV) is an example of an emerging human pathogen with increased and rapid global spread. Although an acute CHIKV infection is rarely fatal, many patients suffer from debilitating chronic arthralgia for years. Antivirals against chikungunya and other alphaviruses have been identified in vitro, but to date none have been shown to be efficacious and have been licensed for human use. Here, we investigated a small molecule, berberine chloride (BBC), and showed that it inhibited infectious virus production by several alphaviruses including CHIKV. BBC acted on a late step in the alphavirus exit pathway, namely the formation of the nucleocapsid containing the infectious viral RNA. Better understanding of nucleocapsid formation and its inhibition by BBC will provide important information on the mechanisms of infectious alphavirus production and may enable their future targeting in antiviral strategies.
format	article
author	Judy J. Wan Rebecca S. Brown Margaret Kielian
author_facet	Judy J. Wan Rebecca S. Brown Margaret Kielian
author_sort	Judy J. Wan
title	Berberine Chloride is an Alphavirus Inhibitor That Targets Nucleocapsid Assembly
title_short	Berberine Chloride is an Alphavirus Inhibitor That Targets Nucleocapsid Assembly
title_full	Berberine Chloride is an Alphavirus Inhibitor That Targets Nucleocapsid Assembly
title_fullStr	Berberine Chloride is an Alphavirus Inhibitor That Targets Nucleocapsid Assembly
title_full_unstemmed	Berberine Chloride is an Alphavirus Inhibitor That Targets Nucleocapsid Assembly
title_sort	berberine chloride is an alphavirus inhibitor that targets nucleocapsid assembly
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/f23789612e504d87af66ef62aa096096
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_version_	1718427128005918720

Berberine Chloride is an Alphavirus Inhibitor That Targets Nucleocapsid Assembly

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