Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1997 Jul;71(7):4985–4989. doi: 10.1128/jvi.71.7.4985-4989.1997

GB virus B and hepatitis C virus NS3 serine proteases share substrate specificity.

E Scarselli 1, A Urbani 1, A Sbardellati 1, L Tomei 1, R De Francesco 1, C Traboni 1
PMCID: PMC191730  PMID: 9188562

Abstract

GB virus B (GBV-B) is a recently discovered virus responsible for hepatitis in tamarins (Saguinus species). GBV-B belongs to the Flaviviridae family and is closely related to the human pathogen hepatitis C virus (HCV). Nonstructural protein 3 (NS3) of HCV has been shown to encompass a serine protease domain required for viral maturation. GBV-B and HCV share only about 30% of the amino acid sequence within the NS3 protease domain. The catalytic triad is conserved, and the residue Phe-154, presumed to be a crucial amino acid for determining the S1 specificity pocket of the HCV NS3 protease, is also conserved. We have expressed a synthetic gene encoding the GBV-B NS3 protease domain in Escherichia coli and have characterized the purified recombinant protein for its activity on HCV substrates. We have shown that the NS3 region of the GBV-B genome actually encodes a serine protease that, despite the low sequence homology, shares substrate specificity with the HCV NS3 protease.

Full Text

The Full Text of this article is available as a PDF (663.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abe K., Inchauspe G., Shikata T., Prince A. M. Three different patterns of hepatitis C virus infection in chimpanzees. Hepatology. 1992 Apr;15(4):690–695. doi: 10.1002/hep.1840150423. [DOI] [PubMed] [Google Scholar]
  2. Darke P. L., Chen E., Hall D. L., Sardana M. K., Veloski C. A., LaFemina R. L., Shafer J. A., Kuo L. C. Purification of active herpes simplex virus-1 protease expressed in Escherichia coli. J Biol Chem. 1994 Jul 22;269(29):18708–18711. [PubMed] [Google Scholar]
  3. De Francesco R., Urbani A., Nardi M. C., Tomei L., Steinkühler C., Tramontano A. A zinc binding site in viral serine proteinases. Biochemistry. 1996 Oct 15;35(41):13282–13287. doi: 10.1021/bi9616458. [DOI] [PubMed] [Google Scholar]
  4. Deinhardt F., Holmes A. W., Capps R. B., Popper H. Studies on the transmission of human viral hepatitis to marmoset monkeys. I. Transmission of disease, serial passages, and description of liver lesions. J Exp Med. 1967 Apr 1;125(4):673–688. doi: 10.1084/jem.125.4.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DiIanni C. L., Mapelli C., Drier D. A., Tsao J., Natarajan S., Riexinger D., Festin S. M., Bolgar M., Yamanaka G., Weinheimer S. P. In vitro activity of the herpes simplex virus type 1 protease with peptide substrates. J Biol Chem. 1993 Dec 5;268(34):25449–25454. [PubMed] [Google Scholar]
  6. Failla C. M., Pizzi E., De Francesco R., Tramontano A. Redesigning the substrate specificity of the hepatitis C virus NS3 protease. Fold Des. 1996;1(1):35–42. [PubMed] [Google Scholar]
  7. Failla C., Tomei L., De Francesco R. An amino-terminal domain of the hepatitis C virus NS3 protease is essential for interaction with NS4A. J Virol. 1995 Mar;69(3):1769–1777. doi: 10.1128/jvi.69.3.1769-1777.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Failla C., Tomei L., De Francesco R. Both NS3 and NS4A are required for proteolytic processing of hepatitis C virus nonstructural proteins. J Virol. 1994 Jun;68(6):3753–3760. doi: 10.1128/jvi.68.6.3753-3760.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Farci P., London W. T., Wong D. C., Dawson G. J., Vallari D. S., Engle R., Purcell R. H. The natural history of infection with hepatitis C virus (HCV) in chimpanzees: comparison of serologic responses measured with first- and second-generation assays and relationship to HCV viremia. J Infect Dis. 1992 Jun;165(6):1006–1011. doi: 10.1093/infdis/165.6.1006. [DOI] [PubMed] [Google Scholar]
  10. Grakoui A., McCourt D. W., Wychowski C., Feinstone S. M., Rice C. M. Characterization of the hepatitis C virus-encoded serine proteinase: determination of proteinase-dependent polyprotein cleavage sites. J Virol. 1993 May;67(5):2832–2843. doi: 10.1128/jvi.67.5.2832-2843.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kim J. L., Morgenstern K. A., Lin C., Fox T., Dwyer M. D., Landro J. A., Chambers S. P., Markland W., Lepre C. A., O'Malley E. T. Crystal structure of the hepatitis C virus NS3 protease domain complexed with a synthetic NS4A cofactor peptide. Cell. 1996 Oct 18;87(2):343–355. doi: 10.1016/s0092-8674(00)81351-3. [DOI] [PubMed] [Google Scholar]
  12. Lin C., Thomson J. A., Rice C. M. A central region in the hepatitis C virus NS4A protein allows formation of an active NS3-NS4A serine proteinase complex in vivo and in vitro. J Virol. 1995 Jul;69(7):4373–4380. doi: 10.1128/jvi.69.7.4373-4380.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Love R. A., Parge H. E., Wickersham J. A., Hostomsky Z., Habuka N., Moomaw E. W., Adachi T., Hostomska Z. The crystal structure of hepatitis C virus NS3 proteinase reveals a trypsin-like fold and a structural zinc binding site. Cell. 1996 Oct 18;87(2):331–342. doi: 10.1016/s0092-8674(00)81350-1. [DOI] [PubMed] [Google Scholar]
  14. 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. Genomic organization of GB viruses A and B: two new members of the Flaviviridae associated with GB agent hepatitis. J Virol. 1995 Sep;69(9):5621–5630. doi: 10.1128/jvi.69.9.5621-5630.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ohba K., Mizokami M., Lau J. Y., Orito E., Ikeo K., Gojobori T. Evolutionary relationship of hepatitis C, pesti-, flavi-, plantviruses, and newly discovered GB hepatitis agents. FEBS Lett. 1996 Jan 15;378(3):232–234. doi: 10.1016/0014-5793(95)01441-1. [DOI] [PubMed] [Google Scholar]
  16. Pilot-Matias T. J., Muerhoff A. S., Simons J. N., Leary T. P., Buijk S. L., Chalmers M. L., Erker J. C., Dawson G. J., Desai S. M., Mushahwar I. K. Identification of antigenic regions in the GB hepatitis viruses GBV-A, GBV-B, and GBV-C. J Med Virol. 1996 Apr;48(4):329–338. doi: 10.1002/(SICI)1096-9071(199604)48:4<329::AID-JMV6>3.0.CO;2-9. [DOI] [PubMed] [Google Scholar]
  17. Pizzi E., Tramontano A., Tomei L., La Monica N., Failla C., Sardana M., Wood T., De Francesco R. Molecular model of the specificity pocket of the hepatitis C virus protease: implications for substrate recognition. Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):888–892. doi: 10.1073/pnas.91.3.888. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sardana V. V., Wolfgang J. A., Veloski C. A., Long W. J., LeGrow K., Wolanski B., Emini E. A., LaFemina R. L. Peptide substrate cleavage specificity of the human cytomegalovirus protease. J Biol Chem. 1994 May 20;269(20):14337–14340. [PubMed] [Google Scholar]
  20. Schaluder G. G., Dawson G. J., Simons J. N., Pilot-Matias T. J., Gutierrez R. A., Heynen C. A., Knigge M. F., Kurpiewski G. S., Buijk S. L., Leary T. P. Molecular and serologic analysis in the transmission of the GB hepatitis agents. J Med Virol. 1995 May;46(1):81–90. doi: 10.1002/jmv.1890460117. [DOI] [PubMed] [Google Scholar]
  21. Schlauder G. G., Pilot-Matias T. J., Gabriel G. S., Simons J. N., Muerhoff A. S., Dawson G. J., Mushahwar I. K. Origin of GB-hepatitis viruses. Lancet. 1995 Aug 12;346(8972):447–448. doi: 10.1016/s0140-6736(95)92821-9. [DOI] [PubMed] [Google Scholar]
  22. Simons J. N., Leary T. P., Dawson G. J., Pilot-Matias T. J., Muerhoff A. S., Schlauder G. G., Desai S. M., Mushahwar I. K. Isolation of novel virus-like sequences associated with human hepatitis. Nat Med. 1995 Jun;1(6):564–569. doi: 10.1038/nm0695-564. [DOI] [PubMed] [Google Scholar]
  23. Simons J. N., Pilot-Matias T. J., Leary T. P., Dawson G. J., Desai S. M., Schlauder G. G., Muerhoff A. S., Erker J. C., Buijk S. L., Chalmers M. L. Identification of two flavivirus-like genomes in the GB hepatitis agent. Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3401–3405. doi: 10.1073/pnas.92.8.3401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Steinkühler C., Tomei L., De Francesco R. In vitro activity of hepatitis C virus protease NS3 purified from recombinant Baculovirus-infected Sf9 cells. J Biol Chem. 1996 Mar 15;271(11):6367–6373. doi: 10.1074/jbc.271.11.6367. [DOI] [PubMed] [Google Scholar]
  25. Steinkühler C., Urbani A., Tomei L., Biasiol G., Sardana M., Bianchi E., Pessi A., De Francesco R. Activity of purified hepatitis C virus protease NS3 on peptide substrates. J Virol. 1996 Oct;70(10):6694–6700. doi: 10.1128/jvi.70.10.6694-6700.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stemmer W. P., Crameri A., Ha K. D., Brennan T. M., Heyneker H. L. Single-step assembly of a gene and entire plasmid from large numbers of oligodeoxyribonucleotides. Gene. 1995 Oct 16;164(1):49–53. doi: 10.1016/0378-1119(95)00511-4. [DOI] [PubMed] [Google Scholar]
  27. Tabor S., Richardson C. C. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. doi: 10.1073/pnas.82.4.1074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Takamizawa A., Mori C., Fuke I., Manabe S., Murakami S., Fujita J., Onishi E., Andoh T., Yoshida I., Okayama H. Structure and organization of the hepatitis C virus genome isolated from human carriers. J Virol. 1991 Mar;65(3):1105–1113. doi: 10.1128/jvi.65.3.1105-1113.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tomei L., Failla C., Santolini E., De Francesco R., La Monica N. NS3 is a serine protease required for processing of hepatitis C virus polyprotein. J Virol. 1993 Jul;67(7):4017–4026. doi: 10.1128/jvi.67.7.4017-4026.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zuckerman A. J. The new GB hepatitis viruses. Lancet. 1995 Jun 10;345(8963):1453–1454. doi: 10.1016/s0140-6736(95)91032-8. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES