1887

Journal of General Virology header logo

Volume 89, Issue 8

A comparative cell biological analysis reveals only limited functional homology between the NS5A proteins of hepatitis C virus and GB virus B Free

Abstract

GB virus B (GBV-B) is the closest relative to hepatitis C virus (HCV) with which it shares a common genome organization, however, unlike HCV in humans, it generally causes an acute resolving hepatitis in New World monkeys. It is important to understand the factors regulating the different disease profiles of the two viruses and in this regard, as well as playing a key role in viral RNA replication, the HCV NS5A non-structural protein modulates a variety of host-cell signalling pathways. We have shown previously that HCV NS5A, expressed either alone, or in the context of the complete polyprotein, inhibits the Ras-extracellular-signal-regulated kinase (Erk) pathway and activates the phosphoinositide 3-kinase (PI3K) pathway. In this report, we investigate whether these functions are shared by GBV-B NS5A. Immunofluorescence analysis revealed that a C-terminally FLAG-tagged GBV-B NS5A exhibited a punctate cytoplasmic distribution. However, unlike HCV NS5A, the GBV-B protein did not partially co-localize with early endosomes. Utilizing a transient luciferase reporter system, we observed that GBV-B NS5A failed to inhibit Ras–Erk signalling, however GBV-B NS5A expression did result in the elevation of -catenin-dependent transcription via activation of the PI3K pathway. These effects of GBV-B and HCV NS5A on the PI3K and Ras–Erk pathways were confirmed in cells harbouring subgenomic replicons derived from the two viruses. Based on these data we speculate that the differential effects of the two NS5A proteins on cellular signalling pathways may contribute to the differences in the natural history of the two viruses.

  • Received:
  • Accepted:
  • Published Online:
SGM
Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.2008/001131-0
2008-08-01
2024-04-18
Loading full text...

Full text loading...

/deliver/fulltext/jgv/89/8/1911.html?itemId=/content/journal/jgv/10.1099/vir.0.2008/001131-0&mimeType=html&fmt=ahah

References

  1. Beames B., Chavez D., Lanford R. E. 2001; GB virus B as a model for hepatitis C virus. ILAR J 42:152–160 [CrossRef]
     [Google Scholar]
  2. Brass V., Bieck E., Montserret R., Wolk B., Hellings J. A., Blum H. E., Penin F., Moradpour D. 2002; An amino-terminal amphipathic α -helix mediates membrane association of the hepatitis C virus nonstructural protein 5A. J Biol Chem 277:8130–8139 [CrossRef]
     [Google Scholar]
  3. Brass V., Pal Z., Sapay N., Deleage G., Blum H. E., Penin F., Moradpour D. 2007; Conserved determinants for membrane association of nonstructural protein 5A from hepatitis C virus and related viruses. J Virol 81:2745–2757 [CrossRef]
     [Google Scholar]
  4. Bright H., Carroll A. R., Watts P. A., Fenton R. J. 2004; Development of a GB virus B marmoset model and its validation with a novel series of hepatitis C virus NS3 protease inhibitors. J Virol 78:2062–2071 [CrossRef]
     [Google Scholar]
  5. Buckwold V. E., Collins B., Hogan P., Rippeon S., Wei J. 2005; Investigation into the ability of GB virus B to replicate in various immortalized cell lines. Antiviral Res 66:165–168 [CrossRef]
     [Google Scholar]
  6. Bukh J., Apgar C. L., Govindarajan S., Purcell R. H. 2001; Host range studies of GB virus-B hepatitis agent, the closest relative of hepatitis C virus, in New World monkeys and chimpanzees. J Med Virol 65:694–697 [CrossRef]
     [Google Scholar]
  7. Chen Z., Benureau Y., Rijnbrand R., Yi J., Wang T., Warter L., Lanford R. E., Weinman S. A., Lemon S. M. other authors 2007; GB virus B disrupts RIG-I signaling by NS3/4A-mediated cleavage of the adaptor protein MAVS. J Virol 81:964–976 [CrossRef]
     [Google Scholar]
  8. Desbois-Mouthon C., Eggelpoel M. J. B. V., Beurel E., Boissan M., Delelo R., Cadoret A., Capeau J. 2002; Dysregulation of glycogen synthase kinase-3 β signaling in hepatocellular carcinoma cells. Hepatology 36:1528–1536 [CrossRef]
     [Google Scholar]
  9. Doble B. W., Woodgett J. R. 2003; GSK-3: tricks of the trade for a multi-tasking kinase. J Cell Sci 116:1175–1186 [CrossRef]
     [Google Scholar]
  10. Egger D., Wolk 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]
     [Google Scholar]
  11. Georgopoulou U., Caravokiri K., Mavromara P. 2003; Suppression of the ERK1/2 signaling pathway from HCV NS5A protein expressed by herpes simplex recombinant viruses. Arch Virol 148:237–251 [CrossRef]
     [Google Scholar]
  12. Green S., Issemann I., Sheer E. 1988; A versatile in vivo and in vitro eukaryotic expression vector for protein engineering. Nucleic Acids Res 16:369 [CrossRef]
     [Google Scholar]
  13. He Y., Nakao H. H., Tan S. L., Polyak P. J., Neddermann P., Vijaysri S., Jacobs B. L., Katze M. G. 2002; Subversion of cell signaling pathways by hepatitis C virus nonstructural 5A protein via interaction with Grb2 and P85 phosphatidylinositol 3-kinase. J Virol 76:9207–9217 [CrossRef]
     [Google Scholar]
  14. Huang Y., Chen X. C., Konduri M., Fomina N., Lu J., Jin L., Kolykhalov A., Tan S. L. 2006; Mechanistic link between the anti-HCV effect of interferon gamma and control of viral replication by a Ras-MAPK signaling cascade. Hepatology 43:81–90 [CrossRef]
     [Google Scholar]
  15. Jacob J. R., Lin K. C., Tennant B. C., Mansfield K. G. 2004; GB virus B infection of the common marmoset ( Callithrix jacchus ) and associated liver pathology. J Gen Virol 85:2525–2533 [CrossRef]
     [Google Scholar]
  16. Krieger N., Lohmann V., Bartenschlager R. 2001; Enhancement of hepatitis C virus RNA replication by cell culture-adaptive mutations. J Virol 75:4614–4624 [CrossRef]
     [Google Scholar]
  17. Lanford R. E., Chavez D., Notvall L., Brasky K. M. 2003; Comparison of tamarins and marmosets as hosts for GBV-B infections and the effect of immunosuppression on duration of viremia. Virology 311:72–80 [CrossRef]
     [Google Scholar]
  18. Lee C. J., Liao C. L., Lin Y. L. 2005; Flavivirus activates phosphatidylinositol 3-kinase signaling to block caspase-dependent apoptotic cell death at the early stage of virus infection. J Virol 79:8388–8399 [CrossRef]
     [Google Scholar]
  19. Lohmann V., Korner F., Koch J. O., Herian U., Theilmann L., Bartenschlager R. 1999; Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line. Science 285:110–113 [CrossRef]
     [Google Scholar]
  20. Macdonald A., Harris M. 2004; Hepatitis C virus NS5A: tales of a promiscuous protein. J Gen Virol 85:2485–2502 [CrossRef]
     [Google Scholar]
  21. Macdonald A., Crowder K., Street A., McCormick C., Saksela K., Harris M. 2003; The hepatitis C virus NS5A protein inhibits activating protein-1 (AP1) function by perturbing Ras–ERK pathway signalling. J Biol Chem 278:17775–17784 [CrossRef]
     [Google Scholar]
  22. Macdonald A., Crowder K., Street A., McCormick C., Harris M. 2004; The hepatitis C virus NS5A protein binds to members of the Src family of tyrosine kinases and regulates kinase activity. J Gen Virol 85:721–729 [CrossRef]
     [Google Scholar]
  23. Macdonald A., Chan J. K., Harris M. 2005a; Perturbation of epidermal growth factor receptor complex formation and Ras signalling in cells harbouring the hepatitis C virus subgenomic replicon. J Gen Virol 86:1027–1033 [CrossRef]
     [Google Scholar]
  24. Macdonald A., Mazaleyrat S., McCormick C., Street A., Burgoyne N. J., Jackson R. M., Cazeaux V., Shelton H., Saksela K., Harris M. 2005b; Further studies on hepatitis C virus NS5A–SH3 domain interactions: identification of residues critical for binding and implications for viral RNA replication and modulation of cell signalling. J Gen Virol 86:1035–1044 [CrossRef]
     [Google Scholar]
  25. Marshall A., Rushbrook S., Davies S. E., Morris L. S., Scott I. S., Vowler S. L., Coleman N., Alexander G. 2005; Relation between hepatocyte G1 arrest, impaired hepatic regeneration, and fibrosis in chronic hepatitis C virus infection. Gastroenterology 128:33–42 [CrossRef]
     [Google Scholar]
  26. Martin A., Bodola F., Sangar D. V., Goettge K., Popov V., Rijnbrand R., Lanford R. E., Lemon S. M. 2003; Chronic hepatitis associated with GB virus B persistence in a tamarin after intrahepatic inoculation of synthetic viral RNA. Proc Natl Acad Sci U S A 100:9962–9967 [CrossRef]
     [Google Scholar]
  27. McCormick C. J., Maucourant S., Griffin S., Rowlands D. J., Harris M. 2006; Tagging of NS5A expressed from a functional hepatitis C virus replicon. J Gen Virol 87:635–640 [CrossRef]
     [Google Scholar]
  28. Mottola G., Cardinali G., Ceccacci A., Trozzi C., Bartholomew L., Torrisi M. R., Pedrazzini E., Bonatti S., Migliaccio G. 2002; Hepatitis C virus nonstructural proteins are localized in a modified endoplasmic reticulum of cells expressing viral subgenomic replicons. Virology 293:31–43 [CrossRef]
     [Google Scholar]
  29. Muerhoff A. S., Leary T. P., Simons J. N., Pilot-Matias T. J., Dawson G. J., Erker J. C., Chalmers M. L., Schlauder G. G., Desai S. M., Mushahwar I. K. 1995; Genomic organization of GB viruses A and B: two new members of the Flaviviridae associated with GB agent hepatitis. J Virol 69:5621–5630
     [Google Scholar]
  30. Nam J. H., Faulk K., Engle R. E., Govindarajan S., St C. M., Bukh J. 2004; In vivo analysis of the 3′ untranslated region of GB virus B after in vitro mutagenesis of an infectious cDNA clone: persistent infection in a transfected tamarin. J Virol 78:9389–9399 [CrossRef]
     [Google Scholar]
  31. Ohba K., Mizokami M., Lau J. Y., Orito E., Ikeo K., Gojobori T. 1996; Evolutionary relationship of hepatitis C, pesti-, flavi-, plant viruses, and newly discovered GB hepatitis agents. FEBS Lett 378:232–234 [CrossRef]
     [Google Scholar]
  32. Stasyk T., Schiefermeier N., Skvortsov S., Zwierzina H., Peranen J., Bonn G. K., Huber L. A. 2007; Identification of endosomal EGF-receptor signaling targets by functional organelle proteomics. Mol Cell Proteomics 6:908–922 [CrossRef]
     [Google Scholar]
  33. Stone M., Jia S., Do H. W., Meyer T., Konan K. V. 2007; Participation of Rab5, an early endosome protein, in hepatitis C virus RNA replication machinery. J Virol 81:4551–4563 [CrossRef]
     [Google Scholar]
  34. Street A., Macdonald A., Crowder K., Harris M. 2004; The hepatitis C virus NS5A protein activates a phosphoinositide 3-kinase dependent survival signalling cascade. J Biol Chem 279:12232–12241 [CrossRef]
     [Google Scholar]
  35. Street A., Macdonald A., McCormick C., Harris M. 2005; Hepatitis C virus NS5A-mediated activation of phosphoinositide 3-kinase results in stabilization of cellular β -catenin and stimulation of β -catenin-responsive transcription. J Virol 79:5006–5016 [CrossRef]
     [Google Scholar]
  36. Talarmin H., Rescan C., Cariou S., Glaise D., Zanninelli G., Bilodeau M., Loyer P., Guguen-Guillouzo C., Baffet G. 1999; The mitogen-activated protein kinase kinase/extracellular signal-regulated kinase cascade activation is a key signalling pathway involved in the regulation of G1 phase progression in proliferating hepatocytes. Mol Cell Biol 19:6003–6011
     [Google Scholar]
  37. Tan S. L., Nakao H., He Y. P., Vijaysri S., Neddermann P., Jacobs B. L., Mayer B. J., Katze M. G. 1999; NS5A, a nonstructural protein of hepatitis C virus, binds growth factor receptor-bound protein 2 adaptor protein in a Src homology 3 domain/ligand-dependent manner and perturbs mitogenic signaling. Proc Natl Acad Sci U S A 96:5533–5538 [CrossRef]
     [Google Scholar]
  38. Tang W., Lazaro C. A., Campbell J. S., Parks W. T., Katze M. G., Fausto N. 2007; Responses of nontransformed human hepatocytes to conditional expression of full-length hepatitis C virus open reading frame. Am J Pathol 171:1831–1846 [CrossRef]
     [Google Scholar]
  39. Tellinghuisen T. L., Marcotrigiano J., Gorbalenya A. E., Rice C. M. 2004; The NS5A protein of hepatitis C virus is a zinc metalloprotein. J Biol Chem 279:48576–48587 [CrossRef]
     [Google Scholar]
  40. Vieira A. V., Lamaze C., Schmid S. L. 1996; Control of EGF receptor signaling by clathrin-mediated endocytosis. Science 274:2086–2089 [CrossRef]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.2008/001131-0
Loading
A comparative cell biological analysis reveals only limited functional homology between the NS5A proteins of hepatitis C virus and GB virus B
J. Gen. Virol. 89, 1911 (2008); https://doi.org/10.1099/vir.0.2008/001131-0
/content/journal/jgv/10.1099/vir.0.2008/001131-0
/content/journal/jgv/10.1099/vir.0.2008/001131-0
Loading

Data & Media loading...

Most cited Most Cited RSS feed

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