Analysis of functional differences between hepatitis C virus NS5A of genotypes 1-7 in infectious cell culture systems.

Hepatitis C virus (HCV) is an important cause of chronic liver disease. Several highly diverse HCV genotypes exist with potential key functional differences. The HCV NS5A protein was associated with response to interferon (IFN)-α based therapy, and is a primary target of currently developed directly...

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Autores principales:	Troels K H Scheel, Jannick Prentoe, Thomas H R Carlsen, Lotte S Mikkelsen, Judith M Gottwein, Jens Bukh
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/6b044b41d51e498fb349f3840a25bf5a
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spelling	oai:doaj.org-article:6b044b41d51e498fb349f3840a25bf5a2021-11-18T06:04:23ZAnalysis of functional differences between hepatitis C virus NS5A of genotypes 1-7 in infectious cell culture systems.1553-73661553-737410.1371/journal.ppat.1002696https://doaj.org/article/6b044b41d51e498fb349f3840a25bf5a2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22654662/pdf/?tool=EBIhttps://doaj.org/toc/1553-7366https://doaj.org/toc/1553-7374Hepatitis C virus (HCV) is an important cause of chronic liver disease. Several highly diverse HCV genotypes exist with potential key functional differences. The HCV NS5A protein was associated with response to interferon (IFN)-α based therapy, and is a primary target of currently developed directly-acting antiviral compounds. NS5A is important for replication and virus production, but has not been studied for most HCV genotypes. We studied the function of NS5A using infectious NS5A genotype 1-7 cell culture systems, and through reverse genetics demonstrated a universal importance of the amphipathic alpha-helix, domain I and II and the low-complexity sequence (LCS) I for HCV replication; the replicon-enhancing LCSI mutation S225P attenuated all genotypes. Mutation of conserved prolines in LCSII led to minor reductions in virus production for the JFH1(genotype 2a) NS5A recombinant, but had greater effects on other isolates; replication was highly attenuated for ED43(4a) and QC69(7a) recombinants. Deletion of the conserved residues 414-428 in domain III reduced virus production for most recombinants but not JFH1(2a). Reduced virus production was linked to attenuated replication in all cases, but ED43(4a) and SA13(5a) also displayed impaired particle assembly. Compared to the original H77C(1a) NS5A recombinant, the changes in LCSII and domain III reduced the amounts of NS5A present. For H77C(1a) and TN(1a) NS5A recombinants, we observed a genetic linkage between NS5A and p7, since introduced changes in NS5A led to changes in p7 and vice versa. Finally, NS5A function depended on genotype-specific residues in domain I, as changing genotype 2a-specific residues to genotype 1a sequence and vice versa led to highly attenuated mutants. In conclusion, this study identified NS5A genetic elements essential for all major HCV genotypes in infectious cell culture systems. Genotype- or isolate- specific NS5A functional differences were identified, which will be important for understanding of HCV NS5A function and therapeutic targeting.Troels K H ScheelJannick PrentoeThomas H R CarlsenLotte S MikkelsenJudith M GottweinJens BukhPublic Library of Science (PLoS)articleImmunologic diseases. AllergyRC581-607Biology (General)QH301-705.5ENPLoS Pathogens, Vol 8, Iss 5, p e1002696 (2012)
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 Troels K H Scheel Jannick Prentoe Thomas H R Carlsen Lotte S Mikkelsen Judith M Gottwein Jens Bukh Analysis of functional differences between hepatitis C virus NS5A of genotypes 1-7 in infectious cell culture systems.
description	Hepatitis C virus (HCV) is an important cause of chronic liver disease. Several highly diverse HCV genotypes exist with potential key functional differences. The HCV NS5A protein was associated with response to interferon (IFN)-α based therapy, and is a primary target of currently developed directly-acting antiviral compounds. NS5A is important for replication and virus production, but has not been studied for most HCV genotypes. We studied the function of NS5A using infectious NS5A genotype 1-7 cell culture systems, and through reverse genetics demonstrated a universal importance of the amphipathic alpha-helix, domain I and II and the low-complexity sequence (LCS) I for HCV replication; the replicon-enhancing LCSI mutation S225P attenuated all genotypes. Mutation of conserved prolines in LCSII led to minor reductions in virus production for the JFH1(genotype 2a) NS5A recombinant, but had greater effects on other isolates; replication was highly attenuated for ED43(4a) and QC69(7a) recombinants. Deletion of the conserved residues 414-428 in domain III reduced virus production for most recombinants but not JFH1(2a). Reduced virus production was linked to attenuated replication in all cases, but ED43(4a) and SA13(5a) also displayed impaired particle assembly. Compared to the original H77C(1a) NS5A recombinant, the changes in LCSII and domain III reduced the amounts of NS5A present. For H77C(1a) and TN(1a) NS5A recombinants, we observed a genetic linkage between NS5A and p7, since introduced changes in NS5A led to changes in p7 and vice versa. Finally, NS5A function depended on genotype-specific residues in domain I, as changing genotype 2a-specific residues to genotype 1a sequence and vice versa led to highly attenuated mutants. In conclusion, this study identified NS5A genetic elements essential for all major HCV genotypes in infectious cell culture systems. Genotype- or isolate- specific NS5A functional differences were identified, which will be important for understanding of HCV NS5A function and therapeutic targeting.
format	article
author	Troels K H Scheel Jannick Prentoe Thomas H R Carlsen Lotte S Mikkelsen Judith M Gottwein Jens Bukh
author_facet	Troels K H Scheel Jannick Prentoe Thomas H R Carlsen Lotte S Mikkelsen Judith M Gottwein Jens Bukh
author_sort	Troels K H Scheel
title	Analysis of functional differences between hepatitis C virus NS5A of genotypes 1-7 in infectious cell culture systems.
title_short	Analysis of functional differences between hepatitis C virus NS5A of genotypes 1-7 in infectious cell culture systems.
title_full	Analysis of functional differences between hepatitis C virus NS5A of genotypes 1-7 in infectious cell culture systems.
title_fullStr	Analysis of functional differences between hepatitis C virus NS5A of genotypes 1-7 in infectious cell culture systems.
title_full_unstemmed	Analysis of functional differences between hepatitis C virus NS5A of genotypes 1-7 in infectious cell culture systems.
title_sort	analysis of functional differences between hepatitis c virus ns5a of genotypes 1-7 in infectious cell culture systems.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/6b044b41d51e498fb349f3840a25bf5a
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Analysis of functional differences between hepatitis C virus NS5A of genotypes 1-7 in infectious cell culture systems.

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