Production of infectious genotype 1b virus particles in cell culture and impairment by replication enhancing mutations.
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Your vote was cast
Thank you for your feedback
Thank you for your feedback
MetadataShow full item record
AbstractWith the advent of subgenomic hepatitis C virus (HCV) replicons, studies of the intracellular steps of the viral replication cycle became possible. These RNAs are capable of self-amplification in cultured human hepatoma cells, but save for the genotype 2a isolate JFH-1, efficient replication of these HCV RNAs requires replication enhancing mutations (REMs), previously also called cell culture adaptive mutations. These mutations cluster primarily in the central region of non-structural protein 5A (NS5A), but may also reside in the NS3 helicase domain or at a distinct position in NS4B. Most efficient replication has been achieved by combining REMs residing in NS3 with distinct REMs located in NS4B or NS5A. However, in spite of efficient replication of HCV genomes containing such mutations, they do not support production of infectious virus particles. By using the genotype 1b isolate Con1, in this study we show that REMs interfere with HCV assembly. Strongest impairment of virus formation was found with REMs located in the NS3 helicase (E1202G and T1280I) as well as NS5A (S2204R), whereas a highly adaptive REM in NS4B still allowed virus production although relative levels of core release were also reduced. We also show that cells transfected with the Con1 wild type genome or the genome containing the REM in NS4B release HCV particles that are infectious both in cell culture and in vivo. Our data provide an explanation for the in vitro and in vivo attenuation of cell culture adapted HCV genomes and may open new avenues for the development of fully competent culture systems covering the therapeutically most relevant HCV genotypes.
CitationProduction of infectious genotype 1b virus particles in cell culture and impairment by replication enhancing mutations. 2009, 5 (6):e1000475 PLoS Pathog.
The following license files are associated with this item:
- Mutations that permit efficient replication of hepatitis C virus RNA in Huh-7 cells prevent productive replication in chimpanzees.
- Authors: Bukh J, Pietschmann T, Lohmann V, Krieger N, Faulk K, Engle RE, Govindarajan S, Shapiro M, St Claire M, Bartenschlager R
- Issue date: 2002 Oct 29
- Dominant negative effect of wild-type NS5A on NS5A-adapted subgenomic hepatitis C virus RNA replicon.
- Authors: Graziani R, Paonessa G
- Issue date: 2004 Jul
- Trans-complementation of HCV replication by non-structural protein 5A.
- Authors: Tong X, Malcolm BA
- Issue date: 2006 Feb
- Essential role of domain III of nonstructural protein 5A for hepatitis C virus infectious particle assembly.
- Authors: Appel N, Zayas M, Miller S, Krijnse-Locker J, Schaller T, Friebe P, Kallis S, Engel U, Bartenschlager R
- Issue date: 2008 Mar 28
- Cell culture-adaptive NS3 mutations required for the robust replication of genome-length hepatitis C virus RNA.
- Authors: Abe K, Ikeda M, Dansako H, Naka K, Kato N
- Issue date: 2007 Apr