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NS2 is dispensable for efficient assembly of hepatitis C virus-like particles in a bipartite trans-encapsidation system

NS2 is dispensable for efficient assembly of hepatitis C virus-like particles in a bipartite trans-encapsidation system
NS2 is dispensable for efficient assembly of hepatitis C virus-like particles in a bipartite trans-encapsidation system
Infectious hepatitis C virus (HCV) particle production in the genotype 2a JFH-1 based cell culture system involves non-structural proteins in addition to canonical virion components. NS2 has been proposed to act as a protein adaptor, co-ordinating the early stages of virion assembly. However, other studies have identified late-acting roles for this protein, making its precise involvement in infectious particle production unclear. Using a robust, bipartite trans-encapsidation system based upon baculovirus expression of HCV structural proteins, we have generated HCV-like particles (HCV-LP) in the absence of NS2 with overt similarity to wild-type virions. HCV-LP could transduce naive cells with trans-encapsidated sub-genomic replicon RNAs and shared similar biochemical and biophysical properties with JFH-1 HCV. Both genotype 1b and JFH-1 intracellular HCV-LP were produced in the absence of NS2, whereas restoring NS2 to the JFH-1 system dramatically enhanced secreted infectivity, consistent with a late acting role. Our system recapitulated authentic HCV particle assembly via trans-complementation of bicistronic, NS2-deleted chimaeric HCV, which is otherwise deficient in particle production. This closely resembled replicon-mediated NS2 trans-complementation, confirming that baculovirus expression of HCV proteins did not unduly affect particle production. Furthermore, this suggests that separation of structural protein expression from replicating HCV RNAs that are destined to be packaged alleviates an early stage requirement for NS2 during particle formation. This highlights our current lack of understanding of how NS2 mediates assembly, yet comparison of full length and bipartite systems may provide further insight into this process.
0022-1317
Bentham, M. J.
5ae5bc3d-f728-49fe-a8fa-82beff34c54c
Marraikii, N.
d69a6c40-f0a7-48fa-9567-09a4a9effbe2
McCormick, C.J.
0fce14bf-2f67-4d08-991f-114dd1e7f0bd
Rowlands, D. J.
8a1d8cc0-917b-4d65-b445-a02cb0ef2cb1
Griffin, S.
838a9881-0208-4911-bdb9-343bce0f3a55
Bentham, M. J.
5ae5bc3d-f728-49fe-a8fa-82beff34c54c
Marraikii, N.
d69a6c40-f0a7-48fa-9567-09a4a9effbe2
McCormick, C.J.
0fce14bf-2f67-4d08-991f-114dd1e7f0bd
Rowlands, D. J.
8a1d8cc0-917b-4d65-b445-a02cb0ef2cb1
Griffin, S.
838a9881-0208-4911-bdb9-343bce0f3a55

Bentham, M. J., Marraikii, N., McCormick, C.J., Rowlands, D. J. and Griffin, S. (2014) NS2 is dispensable for efficient assembly of hepatitis C virus-like particles in a bipartite trans-encapsidation system. Journal of General Virology. (doi:10.1099/vir.0.068932-0). (PMID:25024280) (In Press)

Record type: Article

Abstract

Infectious hepatitis C virus (HCV) particle production in the genotype 2a JFH-1 based cell culture system involves non-structural proteins in addition to canonical virion components. NS2 has been proposed to act as a protein adaptor, co-ordinating the early stages of virion assembly. However, other studies have identified late-acting roles for this protein, making its precise involvement in infectious particle production unclear. Using a robust, bipartite trans-encapsidation system based upon baculovirus expression of HCV structural proteins, we have generated HCV-like particles (HCV-LP) in the absence of NS2 with overt similarity to wild-type virions. HCV-LP could transduce naive cells with trans-encapsidated sub-genomic replicon RNAs and shared similar biochemical and biophysical properties with JFH-1 HCV. Both genotype 1b and JFH-1 intracellular HCV-LP were produced in the absence of NS2, whereas restoring NS2 to the JFH-1 system dramatically enhanced secreted infectivity, consistent with a late acting role. Our system recapitulated authentic HCV particle assembly via trans-complementation of bicistronic, NS2-deleted chimaeric HCV, which is otherwise deficient in particle production. This closely resembled replicon-mediated NS2 trans-complementation, confirming that baculovirus expression of HCV proteins did not unduly affect particle production. Furthermore, this suggests that separation of structural protein expression from replicating HCV RNAs that are destined to be packaged alleviates an early stage requirement for NS2 during particle formation. This highlights our current lack of understanding of how NS2 mediates assembly, yet comparison of full length and bipartite systems may provide further insight into this process.

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More information

Accepted/In Press date: 14 July 2014
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 368189
URI: https://eprints.soton.ac.uk/id/eprint/368189
ISSN: 0022-1317
PURE UUID: 60007c2c-3bc7-4bf4-8f41-25f6a976f5da
ORCID for C.J. McCormick: ORCID iD orcid.org/0000-0002-6155-9161

Catalogue record

Date deposited: 28 Aug 2014 13:44
Last modified: 20 Jul 2019 00:55

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