NS2 protein of hepatitis C virus interacts with structural and non-structural proteins towards virus assembly.

Growing experimental evidence indicates that, in addition to the physical virion components, the non-structural proteins of hepatitis C virus (HCV) are intimately involved in orchestrating morphogenesis. Since it is dispensable for HCV RNA replication, the non-structural viral protein NS2 is suggest...

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Autores principales:	Costin-Ioan Popescu, Nathalie Callens, Dave Trinel, Philippe Roingeard, Darius Moradpour, Véronique Descamps, Gilles Duverlie, François Penin, Laurent Héliot, Yves Rouillé, Jean Dubuisson
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/aac1d161aed24e28862061d99b3924ef
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spelling	oai:doaj.org-article:aac1d161aed24e28862061d99b3924ef2021-11-18T06:03:36ZNS2 protein of hepatitis C virus interacts with structural and non-structural proteins towards virus assembly.1553-73661553-737410.1371/journal.ppat.1001278https://doaj.org/article/aac1d161aed24e28862061d99b3924ef2011-02-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21347350/?tool=EBIhttps://doaj.org/toc/1553-7366https://doaj.org/toc/1553-7374Growing experimental evidence indicates that, in addition to the physical virion components, the non-structural proteins of hepatitis C virus (HCV) are intimately involved in orchestrating morphogenesis. Since it is dispensable for HCV RNA replication, the non-structural viral protein NS2 is suggested to play a central role in HCV particle assembly. However, despite genetic evidences, we have almost no understanding about NS2 protein-protein interactions and their role in the production of infectious particles. Here, we used co-immunoprecipitation and/or fluorescence resonance energy transfer with fluorescence lifetime imaging microscopy analyses to study the interactions between NS2 and the viroporin p7 and the HCV glycoprotein E2. In addition, we used alanine scanning insertion mutagenesis as well as other mutations in the context of an infectious virus to investigate the functional role of NS2 in HCV assembly. Finally, the subcellular localization of NS2 and several mutants was analyzed by confocal microscopy. Our data demonstrate molecular interactions between NS2 and p7 and E2. Furthermore, we show that, in the context of an infectious virus, NS2 accumulates over time in endoplasmic reticulum-derived dotted structures and colocalizes with both the envelope glycoproteins and components of the replication complex in close proximity to the HCV core protein and lipid droplets, a location that has been shown to be essential for virus assembly. We show that NS2 transmembrane region is crucial for both E2 interaction and subcellular localization. Moreover, specific mutations in core, envelope proteins, p7 and NS5A reported to abolish viral assembly changed the subcellular localization of NS2 protein. Together, these observations indicate that NS2 protein attracts the envelope proteins at the assembly site and it crosstalks with non-structural proteins for virus assembly.Costin-Ioan PopescuNathalie CallensDave TrinelPhilippe RoingeardDarius MoradpourVéronique DescampsGilles DuverlieFrançois PeninLaurent HéliotYves RouilléJean DubuissonPublic Library of Science (PLoS)articleImmunologic diseases. AllergyRC581-607Biology (General)QH301-705.5ENPLoS Pathogens, Vol 7, Iss 2, p e1001278 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
spellingShingle	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5 Costin-Ioan Popescu Nathalie Callens Dave Trinel Philippe Roingeard Darius Moradpour Véronique Descamps Gilles Duverlie François Penin Laurent Héliot Yves Rouillé Jean Dubuisson NS2 protein of hepatitis C virus interacts with structural and non-structural proteins towards virus assembly.
description	Growing experimental evidence indicates that, in addition to the physical virion components, the non-structural proteins of hepatitis C virus (HCV) are intimately involved in orchestrating morphogenesis. Since it is dispensable for HCV RNA replication, the non-structural viral protein NS2 is suggested to play a central role in HCV particle assembly. However, despite genetic evidences, we have almost no understanding about NS2 protein-protein interactions and their role in the production of infectious particles. Here, we used co-immunoprecipitation and/or fluorescence resonance energy transfer with fluorescence lifetime imaging microscopy analyses to study the interactions between NS2 and the viroporin p7 and the HCV glycoprotein E2. In addition, we used alanine scanning insertion mutagenesis as well as other mutations in the context of an infectious virus to investigate the functional role of NS2 in HCV assembly. Finally, the subcellular localization of NS2 and several mutants was analyzed by confocal microscopy. Our data demonstrate molecular interactions between NS2 and p7 and E2. Furthermore, we show that, in the context of an infectious virus, NS2 accumulates over time in endoplasmic reticulum-derived dotted structures and colocalizes with both the envelope glycoproteins and components of the replication complex in close proximity to the HCV core protein and lipid droplets, a location that has been shown to be essential for virus assembly. We show that NS2 transmembrane region is crucial for both E2 interaction and subcellular localization. Moreover, specific mutations in core, envelope proteins, p7 and NS5A reported to abolish viral assembly changed the subcellular localization of NS2 protein. Together, these observations indicate that NS2 protein attracts the envelope proteins at the assembly site and it crosstalks with non-structural proteins for virus assembly.
format	article
author	Costin-Ioan Popescu Nathalie Callens Dave Trinel Philippe Roingeard Darius Moradpour Véronique Descamps Gilles Duverlie François Penin Laurent Héliot Yves Rouillé Jean Dubuisson
author_facet	Costin-Ioan Popescu Nathalie Callens Dave Trinel Philippe Roingeard Darius Moradpour Véronique Descamps Gilles Duverlie François Penin Laurent Héliot Yves Rouillé Jean Dubuisson
author_sort	Costin-Ioan Popescu
title	NS2 protein of hepatitis C virus interacts with structural and non-structural proteins towards virus assembly.
title_short	NS2 protein of hepatitis C virus interacts with structural and non-structural proteins towards virus assembly.
title_full	NS2 protein of hepatitis C virus interacts with structural and non-structural proteins towards virus assembly.
title_fullStr	NS2 protein of hepatitis C virus interacts with structural and non-structural proteins towards virus assembly.
title_full_unstemmed	NS2 protein of hepatitis C virus interacts with structural and non-structural proteins towards virus assembly.
title_sort	ns2 protein of hepatitis c virus interacts with structural and non-structural proteins towards virus assembly.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/aac1d161aed24e28862061d99b3924ef
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NS2 protein of hepatitis C virus interacts with structural and non-structural proteins towards virus assembly.

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