1887

Abstract

Hepatitis C virus (HCV) represents a significant global health burden. Viral replication is thought to occur in close association with remodelled host cell membranes, with non-structural protein 4B (NS4B) being a key player in this process. NS4B is a poorly characterized integral membrane protein, which has been reported to be palmitoylated at its carboxy-terminal end. In order to extend this observation and to establish a functional role for NS4B palmitoylation, we sought to determine the status of this post-translational modification when the protein was expressed as part of a functional viral replicase. We performed direct metabolic labelling and polyethylene glycol-maleimide palmitoylation reporter assays on NS4B expressed in cells containing subgenomic replicons and infectious viral RNA. In a vaccinia virus-based expression system NS4B palmitoylation was detected in a genotype-dependent manner. However, in spite of the high sensitivity of the methods used, no NS4B palmitoylation was found in physiologically more relevant systems. Thus, NS4B palmitoylation is most likely dispensable for HCV RNA replication.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.000111
2015-07-01
2020-01-23
Loading full text...

Full text loading...

/deliver/fulltext/jgv/96/7/1696.html?itemId=/content/journal/jgv/10.1099/vir.0.000111&mimeType=html&fmt=ahah

References

  1. Egger D., Wölk B., Gosert R., Bianchi L., Blum H. E., Moradpour D., Bienz K.. ( 2002; ). Expression of hepatitis C virus proteins induces distinct membrane alterations including a candidate viral replication complex. . J Virol 76:, 5974–5984. [CrossRef] [PubMed]
    [Google Scholar]
  2. Gouttenoire J., Penin F., Moradpour D.. ( 2010; a). Hepatitis C virus nonstructural protein 4B: a journey into unexplored territory. . Rev Med Virol 20:, 117–129. [CrossRef] [PubMed]
    [Google Scholar]
  3. Gouttenoire J., Roingeard P., Penin F., Moradpour D.. ( 2010; b). Amphipathic α-helix AH2 is a major determinant for the oligomerization of hepatitis C virus nonstructural protein 4B. . J Virol 84:, 12529–12537. [CrossRef] [PubMed]
    [Google Scholar]
  4. Herod M. R., Jones D. M., McLauchlan J., McCormick C. J.. ( 2012; ). Increasing rate of cleavage at boundary between non-structural proteins 4B and 5A inhibits replication of hepatitis C virus. . J Biol Chem 287:, 568–580. [CrossRef] [PubMed]
    [Google Scholar]
  5. Krijnse Locker J., Chlanda P., Sachsenheimer T., Brügger B.. ( 2013; ). Poxvirus membrane biogenesis: rupture not disruption. . Cell Microbiol 15:, 190–199. [CrossRef] [PubMed]
    [Google Scholar]
  6. Lakkaraju A. K., Abrami L., Lemmin T., Blaskovic S., Kunz B., Kihara A., Dal Peraro M., van der Goot F. G.. ( 2012; ). Palmitoylated calnexin is a key component of the ribosome-translocon complex. . EMBO J 31:, 1823–1835. [CrossRef] [PubMed]
    [Google Scholar]
  7. Majeau N., Fromentin R., Savard C., Duval M., Tremblay M. J., Leclerc D.. ( 2009; ). Palmitoylation of hepatitis C virus core protein is important for virion production. . J Biol Chem 284:, 33915–33925. [CrossRef] [PubMed]
    [Google Scholar]
  8. McCormick C. J., Challinor L., Macdonald A., Rowlands D. J., Harris M.. ( 2004; ). Introduction of replication-competent hepatitis C virus transcripts using a tetracycline-regulable baculovirus delivery system. . J Gen Virol 85:, 429–439. [CrossRef] [PubMed]
    [Google Scholar]
  9. Paul D., Romero-Brey I., Gouttenoire J., Stoitsova S., Krijnse-Locker J., Moradpour D., Bartenschlager R.. ( 2011; ). NS4B self-interaction through conserved C-terminal elements is required for the establishment of functional hepatitis C virus replication complexes. . J Virol 85:, 6963–6976. [CrossRef] [PubMed]
    [Google Scholar]
  10. Paul D., Hoppe S., Saher G., Krijnse-Locker J., Bartenschlager R.. ( 2013; ). Morphological and biochemical characterization of the membranous hepatitis C virus replication compartment. . J Virol 87:, 10612–10627. [CrossRef] [PubMed]
    [Google Scholar]
  11. Paul D., Madan V., Bartenschlager R.. ( 2014; ). Hepatitis C virus RNA replication and assembly: living on the fat of the land. . Cell Host Microbe 16:, 569–579. [CrossRef] [PubMed]
    [Google Scholar]
  12. Pietschmann T., Kaul A., Koutsoudakis G., Shavinskaya A., Kallis S., Steinmann E., Abid K., Negro F., Dreux M. et al. ( 2006; ). Construction and characterization of infectious intragenotypic and intergenotypic hepatitis C virus chimeras. . Proc Natl Acad Sci U S A 103:, 7408–7413. [CrossRef] [PubMed]
    [Google Scholar]
  13. Romero-Brey I., Merz A., Chiramel A., Lee J. Y., Chlanda P., Haselman U., Santarella-Mellwig R., Habermann A., Hoppe S. et al. ( 2012; ). Three-dimensional architecture and biogenesis of membrane structures associated with hepatitis C virus replication. . PLoS Pathog 8:, e1003056. [CrossRef] [PubMed]
    [Google Scholar]
  14. Roth A. F., Wan J., Bailey A. O., Sun B., Kuchar J. A., Green W. N., Phinney B. S., Yates J. R. III, Davis N. G.. ( 2006; ). Global analysis of protein palmitoylation in yeast. . Cell 125:, 1003–1013. [CrossRef] [PubMed]
    [Google Scholar]
  15. Turnbull A. P., Kümmel D., Prinz B., Holz C., Schultchen J., Lang C., Niesen F. H., Hofmann K. P., Delbrück H. et al. ( 2005; ). Structure of palmitoylated BET3: insights into TRAPP complex assembly and membrane localization. . EMBO J 24:, 875–884. [CrossRef] [PubMed]
    [Google Scholar]
  16. Yu G. Y., Lee K. J., Gao L., Lai M. M.. ( 2006; ). Palmitoylation and polymerization of hepatitis C virus NS4B protein. . J Virol 80:, 6013–6023. [CrossRef] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.000111
Loading
/content/journal/jgv/10.1099/vir.0.000111
Loading

Data & Media loading...

Supplements

Supplementary Data



PDF

Most cited articles

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error