The University of Southampton
University of Southampton Institutional Repository

Expression of hepatitis C virus (HCV) structural proteins in trans facilitates encapsidation and transmission of HCV subgenomic RNA

Expression of hepatitis C virus (HCV) structural proteins in trans facilitates encapsidation and transmission of HCV subgenomic RNA
Expression of hepatitis C virus (HCV) structural proteins in trans facilitates encapsidation and transmission of HCV subgenomic RNA
A characteristic of many positive-strand RNA viruses is that, whilst replication of the viral genome is dependent on the expression of the majority of non-structural proteins in cis, virus particle formation can occur when most or all of the structural proteins are co-expressed in trans. Making use of a recently identified hepatitis C virus (HCV) isolate (JFH1) that can be propagated in tissue culture, this study sought to establish whether this is also the case for hepaciviruses. Stable cell lines containing one of two bicistronic replicons derived from the JFH1 isolate were generated that expressed non-structural proteins NS3-5B or NS2-5B. Release and transmission of these replicons to naïve Huh7 cells could then be demonstrated when baculovirus transduction was used to express the HCV proteins absent from the subgenomic replicons. Transmission could be blocked by a neutralizing antibody targeted at the E2 envelope protein, consistent with this phenomenon occurring via trans-encapsidation of replicon RNA into virus-like particles. Transmission was also dependent on expression of NS2, which was most effective at promoting virus particle formation when expressed in cis on the replicon RNA compared with in trans via baculovirus delivery. Density gradient analysis of the particles revealed the presence of a broad infectious peak between 1.06 and 1.11 g ml(-1), comparable to that seen when propagating full-length virus in tissue culture. In summary, the trans-encapsidation system described offers a complementary and safer approach to study HCV particle formation and transmission in tissue culture.
0022-1317
833-842
Adair, R.
1c436ca7-baa7-4643-b756-8ddf450698e0
Patel, A.H.
3400cd69-5280-4adc-ac54-244b8c5cee17
Corless, L.
5aa0924d-5268-45e4-aa5d-e10a2a71b955
Griffin, S.
838a9881-0208-4911-bdb9-343bce0f3a55
Rowlands, D.J.
a66c545f-90e9-4e77-a0d8-e30ce094bb44
McCormick, C.J.
0fce14bf-2f67-4d08-991f-114dd1e7f0bd
Adair, R.
1c436ca7-baa7-4643-b756-8ddf450698e0
Patel, A.H.
3400cd69-5280-4adc-ac54-244b8c5cee17
Corless, L.
5aa0924d-5268-45e4-aa5d-e10a2a71b955
Griffin, S.
838a9881-0208-4911-bdb9-343bce0f3a55
Rowlands, D.J.
a66c545f-90e9-4e77-a0d8-e30ce094bb44
McCormick, C.J.
0fce14bf-2f67-4d08-991f-114dd1e7f0bd

Adair, R., Patel, A.H., Corless, L., Griffin, S., Rowlands, D.J. and McCormick, C.J. (2009) Expression of hepatitis C virus (HCV) structural proteins in trans facilitates encapsidation and transmission of HCV subgenomic RNA. Journal of General Virology, 90, 833-842. (doi:10.1099/vir.2008.006049-0).

Record type: Article

Abstract

A characteristic of many positive-strand RNA viruses is that, whilst replication of the viral genome is dependent on the expression of the majority of non-structural proteins in cis, virus particle formation can occur when most or all of the structural proteins are co-expressed in trans. Making use of a recently identified hepatitis C virus (HCV) isolate (JFH1) that can be propagated in tissue culture, this study sought to establish whether this is also the case for hepaciviruses. Stable cell lines containing one of two bicistronic replicons derived from the JFH1 isolate were generated that expressed non-structural proteins NS3-5B or NS2-5B. Release and transmission of these replicons to naïve Huh7 cells could then be demonstrated when baculovirus transduction was used to express the HCV proteins absent from the subgenomic replicons. Transmission could be blocked by a neutralizing antibody targeted at the E2 envelope protein, consistent with this phenomenon occurring via trans-encapsidation of replicon RNA into virus-like particles. Transmission was also dependent on expression of NS2, which was most effective at promoting virus particle formation when expressed in cis on the replicon RNA compared with in trans via baculovirus delivery. Density gradient analysis of the particles revealed the presence of a broad infectious peak between 1.06 and 1.11 g ml(-1), comparable to that seen when propagating full-length virus in tissue culture. In summary, the trans-encapsidation system described offers a complementary and safer approach to study HCV particle formation and transmission in tissue culture.

Text
Trans_HCV_paper.pdf - Version of Record
Download (306kB)

More information

Submitted date: 19 July 2008
Published date: 17 February 2009
Organisations: Infection Inflammation & Immunity

Identifiers

Local EPrints ID: 72616
URI: http://eprints.soton.ac.uk/id/eprint/72616
ISSN: 0022-1317
PURE UUID: 7418eacd-6938-4322-86a5-0723986eeade
ORCID for C.J. McCormick: ORCID iD orcid.org/0000-0002-6155-9161

Catalogue record

Date deposited: 18 Feb 2010
Last modified: 14 Mar 2024 02:50

Export record

Altmetrics

Contributors

Author: R. Adair
Author: A.H. Patel
Author: L. Corless
Author: S. Griffin
Author: D.J. Rowlands
Author: C.J. McCormick ORCID iD

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of http://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×