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The Hepatitis C Viruses pp 149–169Cite as

Structure and Function of the Hepatitis C Virus NS3-NS4A Serine Proteinase

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Part of the book series: Current Topics in Microbiology and Immunology ((CT MICROBIOLOGY,volume 242))

Abstract

Hepatitis C virus (HCV) is a relatively young member of the Flaviviridae family that is nowadays recognized as the major etiological agent of both blood-borne and sporadic non-A-non-B hepatitis (HOUGHTON 1996). It is estimated that about 0.5%–1.5% of the total world population is infected with this virus. In many cases, infection with HCV leads to life-threatening disease, such as cirrhosis of the liver and hepatocarcinoma (SIMMONDS et al. 1998). A vaccine against HCV has not yet been discovered, and treatment with interferon is effective only in about 20% of the patients. There is thus a compelling need for a deeper understanding of the HCV life cycle and identification of targets for the development of more effective antiviral treatments.

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References

  • Bartenschlager R, Ahlborn-Laake L, Mous J, Jacobsen H (1993) Nonstructural protein 3 of the hepatitis C virus encodes a serine-type proteinase required for cleavage at the NS3/4 and NS4/5 junctions. J Virol 67:3835–3844

    PubMed  CAS  Google Scholar 

  • Bartenschlager R, Ahlborn Laake L, Mous J, Jacobsen H (1994) Kinetic and structural analyses of hepatitis C virus polyprotein processing. J Virol 68:5045–5055

    PubMed  CAS  Google Scholar 

  • Bartenschlager R, Ahlborn-Laake L, Yasargil K, Mous J, Jacobsen H (1995a) Substrate determinants for cleavage in cis and in trans by the hepatitis C virus NS3 proteinase. J Virol 69:198–205

    PubMed  CAS  Google Scholar 

  • Bartenschlager R, Lohman V, Wilkinson T, Koch JO (1995b) Complex formation between the NS3 serine-type proteinase of the hepatitis C virus and NS4A and its importance for polyprotein matu¬ration. J Virol 69:7519–7528

    PubMed  CAS  Google Scholar 

  • Bartenschlager R (1997) Molecular targets in inhibition of the hepatitis C virus replication. Antiviral Chem Chemother 8:281–301

    CAS  Google Scholar 

  • Bazan FJ, Fletterick RJ (1989) Detection of a trypsin-like serine protease domain in Flavivirus and Pestivirus. Virology 171:637–639

    Article  PubMed  CAS  Google Scholar 

  • Bianchi E, Urbani A, Biasiol G, Brunetti M, Pessi A, De Francesco R, Steinkühler C (1997) Complex formation between the hepatitis C virus serine proteinase and a synthetic NS4A cofactor peptide. Biochemistry 36:7890–7897

    Article  PubMed  CAS  Google Scholar 

  • Butkiewicz NJ, Wendel M, Zhang R, Jubin R, Pichardo J, Smith EB, Hart AM, Ingram R, Durkin J, Mui PW, Murray MG, Ramanathan L, Dasmahapatra B (1996) Enhancement of hepatitis C virus NS3 proteinase activity by association with NS4A-specific synthetic peptides: identification of sequence and critical residues of NS4A for the cofactor activity. Virology 225:328–338

    Article  PubMed  CAS  Google Scholar 

  • Chambers TJ, Weir RC, Grakoui A, McCourt DW, Bazan JF, Fletterick RJ, Rice CM (1990) Evidence that the N-terminal domain of nonstructural protein NS3 from yellow fever virus is a serine protease responsible for site-specific cleavages in the viral polyprotein. Proc Natl Acad Sci USA 87:8898–8902

    Article  PubMed  CAS  Google Scholar 

  • De Francesco R, Urbani A, Nardi MC, Tornei L, Steinkühler C, Tramontano A (1996) A zinc binding site in viral serine proteinases. Biochemistry 35:13282–13287

    Article  PubMed  Google Scholar 

  • Eckart MR, Selby M, Masiarz F, Lee C, Berger K, Crawford K, Kuo G, Houghton M, Choo QL (1993) The hepatitis C virus encodes a serine proteinase involved in processing of the putative nonstructural proteins from the viral polyprotein precursor. Biochem Biophys Res Commun 192:399–406

    Article  PubMed  CAS  Google Scholar 

  • Failla C, Tornei L, De Francesco R (1994) Both NS3 and NS4A are required for proteolytic processing of hepatitis C virus nonstructural proteins. J Virol 68:3753–3760

    PubMed  CAS  Google Scholar 

  • Failla C, Tornei L, De Francesco R (1995) An amino-terminal domain of the hepatitis C virus NS3 proteinase is essential for interaction with NS4A. J Virol 69:1769–1777

    PubMed  CAS  Google Scholar 

  • Failla C, Pizzi E, De Francesco R, Tramontano A (1996) Redesigning the substrate specificity of the hepatitis C virus proteinase. Folding Design 1:35–42

    Article  PubMed  CAS  Google Scholar 

  • Grakoui A, McCourt DW, Wychowski C, Feinstone SM, Rice C (1993) Characterization of the hepatitis C virus-encoded serine proteinase: determination of proteinase-dependent polyprotein cleavage sites. J Virol 67:2832–2843

    PubMed  CAS  Google Scholar 

  • Hahm B, Han DS, Back SH, Song OK, Cho MJ, Kim CJ, Shimotohno K, Jang SK (1995) NS3–4A of hepatitis C virus is a chymotrypsin-like proteinase. J Virol 69:2534–2539

    PubMed  CAS  Google Scholar 

  • Han DS, Hahm B, Rho HM, Jang SK (1995) Identification of the serine proteinase domain in NS3 of the hepatitis C virus. J Gen Virol 76:985–993

    Article  PubMed  CAS  Google Scholar 

  • Hijikata M, Mizushima H, Tanji Y, Komoda Y, Hirowatari Y, Akagi T, Kato N, Kimura K, Shimotohno K (1993a) Proteolytic processing and membrane association of putative nonstructural proteins of hepatitis C virus. Proc Natl Acad Sci USA 90:10733–10737

    Article  Google Scholar 

  • Hijikata M, Mizushima H, Akagi T, Mori S, Kakiuchi N, Kato N, Tanaka T, Kimura K, Shimotohno K (1993b) Two distinct proteinase activities required for the processing of a putative nonstructural precursor protein of hepatitis C virus. J Virol 67:4665–4675

    PubMed  CAS  Google Scholar 

  • Houghton M (1996) Hepatitis C Viruses. In: Fields BN, Knipe DM, Howley PM (eds) Virology, 3rd edn. Lippincott-Raven, Philadelphia, pp 1035–1058

    Google Scholar 

  • Kim JL, Morgenstern KA, Lin C, Fox T, Dwyer MD, Landro JA, Chambers SP, Markland W, Lepre CA, O’Malley ET, Harbeson SL, Rice CM, Murcko MA, Carón PR, Thomson JA (1996) Crystal structure of the hepatitis virus NS3 proteinase domain complexed with a synthetic NS4A cofactor peptide. Cell 87:343–355

    Article  PubMed  CAS  Google Scholar 

  • Koch JO, Lohmann V, Herian U, Bartenschlager R (1996) In vitro studies on the activation of the hepatitis C virus NS3 proteinase by the NS4A cofactor. Virology 221:54–66

    Article  PubMed  CAS  Google Scholar 

  • Koch JO, Bartenschlager R (1997) Determinants of substrate specificity in the NS3 serine proteinase of the hepatitis C virus. Virology 237:78–88

    Article  PubMed  CAS  Google Scholar 

  • Kolykhalov AA, Agapov EV, Rice CM (1994) Specificity of the hepatitis C virus NS3 serine proteinase: effects of substitutions at the 3/4A, 4A/4B, 4B/5A, and 5A/5B cleavage sites on polyprotein processing. J Virol 68:7525–7533

    PubMed  CAS  Google Scholar 

  • Komoda Y, Hijikata M, Sato S, Asabe SI, Kimura K, Shimotohno K (1994b) Substrate requirements of hepatitis C virus serine proteinase for intermolecular polypeptide cleavage in Escherichia coli. J Virol 68:7351–7357

    PubMed  CAS  Google Scholar 

  • Landro JA, Raybuck SA, Luong YPC, O’Malley ET, Harbeson SL, Morgenstern KA, Rao G, Livingston DJ (1997) Mechanistic role of an NS4A peptide cofactor with the truncated NS3 proteinase of hepatitis C virus: elucidation of the NS4A stimulatory effect via kinetic analysis and inhibitor mapping. Biochemistry 36:9340–9348

    Article  PubMed  CAS  Google Scholar 

  • Leinbach SS, Bhat RA, Xia SM, Hum WT, Stauffer B, Davis A, Hung PP, Mizutani S (1994) Substrate specificity of the NS3 serine proteinase of hepatitis C virus as determined by mutagenesis at the NS3/ NS4A junction. Virology 204:163–169

    Article  PubMed  CAS  Google Scholar 

  • Lesk AM, Fordham WD (1996) Conservation and variability in the structure of serine proteinases of the chymotrypsin family. J Mol Biol 258:501–537

    Article  PubMed  CAS  Google Scholar 

  • Lin C, Pragai BM, Grakoui A, Xu J, Rice CM (1994b) Hepatitis C virus NS3 serine proteinase: trans-cleavage requirements and processing kinetics. J Virol 68:8147–8157

    PubMed  CAS  Google Scholar 

  • Lin C, Rice CM (1995) The hepatitis C virus NS3 proteinase and NS4A co-factor: establishment of a cell-free trans-processing assay. Proc Natl Acad Sci USA 92:7622–7626

    Article  PubMed  CAS  Google Scholar 

  • Lin C, Thomson JA, Rice CM (1995) 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 69:4373–4380

    PubMed  CAS  Google Scholar 

  • Love RA, Parge HE, Wickersham JA, Hostomsky Z, Habuka N, Moomaw EW, Adachi T, Homstomska Z (1996) The crystal structure of hepatitis C virus NS3 proteinase reveals a trypsin-like fold and a structural zinc binding site. Cell 87:331–342

    Article  PubMed  CAS  Google Scholar 

  • Manabe S, Fuke I, Tanishita O, Kaji C, Gomi Y, Yoshida S, Mori C, Takamizawa A, Yosida I, Okayama H (1994) Production of nonstructural proteins of hepatitis C virus requires a putative viral proteinase encoded by NS3. Virology 198:636–644

    Article  PubMed  CAS  Google Scholar 

  • Markland W, Petrillo RA, Fitzgibbon M, Fox T, McCarrick R, McQuaid T, Fulghum JR, Chen W, Fleming MA, Thomson JA, Chambers SP (1997) Purification and characterization of the NS3 serine proteinase domain of hepatitis C virus expressed in Saccharomyces cerevisiae. J Gen Virol 78:39–43

    PubMed  CAS  Google Scholar 

  • Miller RH, Purcell RH (1990) Hepatitis C virus shares amino acid sequence similarity with pestiviruses and flaviviruses as well as members of two plant virus supergroups. Proc Natl Acad Sci USA 87:2057–2061

    Article  PubMed  CAS  Google Scholar 

  • Mori A, Yamada K, Kimura J, Koide T, Yuasa S, Yamada E, Miyamura T (1996) Enzymatic characterization of purified NS3 serine proteinase of hepatitis C virus expressed in Escherichia coli. FEBS Lett 378:37–42

    Article  PubMed  CAS  Google Scholar 

  • Neddermann P, Tornei L, Steinkühler C, Gallinari P, Tramontano A, De Francesco R (1997) The nonstructural proteins of the hepatitis C virus: structure and functions. Biol Chem 378:469–476

    PubMed  CAS  Google Scholar 

  • Pieroni L, Santolini E, Fipaldini C, Pacini L, Migliaccio G, La Monica N (1997) In vitro study of the NS2–3 proteinase of hepatitis C virus. J Virol 71:6373–6380

    PubMed  CAS  Google Scholar 

  • Pizzi E, Tramontano A, Tornei L, La Monica N, Failla C, Sardana M, Wood T, De Francesco R (1994) Molecular model of the specificity pocket of the hepatitis C virus proteinase: implications for sub¬strate recognition. Proc Natl Acad Sci USA 91:888–892

    Article  PubMed  CAS  Google Scholar 

  • Reed KE, Grakoui A, Rice CM (1995) Hepatitis C virus-encoded NS2–3 proteinase: cleavage-site mutagenesis and requirements for bimolecular cleavage. J Virol 69:4127–4136

    PubMed  CAS  Google Scholar 

  • Rost B, Casadio R, Fariselli P, Sander C (1995) Transmembrane helices predicted at 95% accuracy. Protein Sci 4:521–533

    Article  PubMed  CAS  Google Scholar 

  • Santolini E, Pacini L, Fipaldini C, Migliaccio G, Monica N (1995) The NS2 protein of hepatitis C virus is a transmembrane polypeptide. J Virol 69:7461–7471

    PubMed  CAS  Google Scholar 

  • Satoh S, Tanji Y, Hijikata M, Kimura K, Shimotohno K (1995) The N-terminal region of hepatitis C virus nonstructural protein 3 (NS3) is essential for stable complex formation with NS4A. J Virol 69:4255–4260

    PubMed  CAS  Google Scholar 

  • Shimizu Y, Yamaji K, Masuho Y, Yokota T, Inoue H, Sudo K, Satoh S, Shimotohno K (1996) Identification of the sequence of NS4A required for enhanced cleavage of the NS5A/5B site by hepatitis C virus NS3 proteinase. J Virol 70:127–132

    PubMed  CAS  Google Scholar 

  • Shoji I, Suzuki T, Chieda S, Sato M, Harada T, Chiba T, Matsuura Y, Miyamura T (1995) Proteolytic activity of NS3 serine proteinase of hepatitis C virus efficiently expressed in Escherichia coli. Hepatology 22:1648–1655

    PubMed  CAS  Google Scholar 

  • Simmonds P, Mutimer D, Follet EAC (1998) Hepatitis C. In: Collier L, Balows A, Sussman M (eds) Microbiology and microbial infections. Oxford University Press, New York

    Google Scholar 

  • Steinkühler C, Tornei L, De Francesco R (1996a) In vitro activity of hepatitis C virus proteinase NS3 purified from recombinant baculovirus-infected Sf9 cells. J Biol Chem 271:6367–6373

    Article  PubMed  Google Scholar 

  • Steinkühler C, Urbani A, Tornei L, Biasiol G, Sardana M, Bianchi E, Pessi A, De Francesco R (1996b) Activity of purified hepatitis C virus proteinase NS3 on peptide substrates. J Virol 70:6694–6700

    PubMed  Google Scholar 

  • Stempniak M, Hostomska Z, Nodes BR, Hostomsky Z (1997) The NS3 proteinase domain of hepatitis C virus is a zinc-containing enzyme. J Virol 71:2881–2886

    PubMed  CAS  Google Scholar 

  • Suzuki T, Sato M, Chieda S, Shoji I, Harada T, Yamakawa Y, Watabe S, Matsuura Y, Miyamura T (1995) In vivo and in vitro trans-cleavage activity of hepatitis C virus serine proteinase expressed by recombinant baculoviruses. J Gen Virol 76:3021–3029

    Article  PubMed  CAS  Google Scholar 

  • Tanji Y, Hijikata M, Hirowatari Y, Shimotohno K (1994a) Hepatitis C virus polyprotein processing: kinetics and mutagenic analysis of serine proteinase-dependent cleavage. J Virol 68:8418–8422

    PubMed  CAS  Google Scholar 

  • Tanji Y, Hijikata M, Hirowatari Y, Shimotohno K (1994b) Identification of the domain required for trans-cleavage activity of hepatitis C viral serine proteinase. Gene 145:215–219

    Article  PubMed  CAS  Google Scholar 

  • Tanji Y, Hijikata M, Satoh S, Kaneko T, Shimotohno K (1995a) Hepatitis C virus-encoded nonstructural protein NS4A has versatile functions in viral protein processing. J Virol 69:1575–1581

    PubMed  CAS  Google Scholar 

  • Tornei L, Failla C, Santolini E, De Francesco R, La Monica N (1993) NS3 is a serine proteinase required for processing of hepatitis C virus polyprotein. J Virol 67:4017–4026

    Google Scholar 

  • Tornei L, Failla C, Vitale RL, Bianchi E, De Francesco R (1996) A centrai hydrophobic domain of the hepatitis C virus NS4A protein is necessary and sufficient for the activation of the NS3 proteinase. J Gen Virol 77:1065–1070

    Article  Google Scholar 

  • Urbani A, Bianchi E, Narjes F, Tramontano A, De Francesco R, Steinkühler C, Pessi A (1997) Substrate specificity of the hepatitis C virus serine proteinase NS3. J Biol Chem 272:9204–9209

    Article  PubMed  CAS  Google Scholar 

  • Urbani A, Bazzo R, Nardi MC, Cicero D, De Francesco R, Steinkühler C, Barbato G (1998) The metal binding site of the hepatitis C virus NS3 proteinase. A spectroscopic study. J Biol Chem 273: 18760–18769

    CAS  Google Scholar 

  • Vishnuvardan D, Kakiuchi N, Urvil PT, Shimotohno K, Kumar PKR, Nishikawa S (1997) Expression of highly active recombinant NS3 proteinase domain of hepatitis C virus in E. coli. FEBS Lett 402: 209–212

    Article  Google Scholar 

  • Yan Y, Li Y, Munshi S, Sardana V, Cole J, Sardana M, Steinkühler C, Tornei L, De Francesco R, Kuo L, Chen Z (1998) Complex of NS3 proteinase and NS4A peptide of BK strain hepatitis C virus: a 2.2A resolution structure in a hexagonal crystal form. Protein Sci 7:837–847

    Article  PubMed  CAS  Google Scholar 

  • Zhang R, Durkin J, Windsor WT, McNemar C, Ramanathan L, Le HV (1997) Probing the substrate specificity of hepatitis C virus NS3 serine proteinase by using synthetic peptides. J Virol 71:6208–6213

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Istuto di Ricerche di Biologia Molecolare (IRBM) “P. Angeletti”, Via Pontina Km 30.600, Pomezia(Rome), 00040, Italy

    R. De Francesco & C. Steinkühler

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  1. R. De Francesco
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  2. C. Steinkühler
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Editors and Affiliations

  1. Division of Digestive Diseases and Genetics — Winship Cancer Center, Emory University School of Medicine, 1639 Pierce Drive, Rm 2101 WMRB, 30322, Atlanta, GA, USA

    Curt H. Hagedorn M.D.

  2. Department of Molecular Microbiology, Washington University School of Medicine, 600 South Euclid Avenue, 63110, St. Louis, MO, USA

    Charles M. Rice M.D.

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De Francesco, R., Steinkühler, C. (2000). Structure and Function of the Hepatitis C Virus NS3-NS4A Serine Proteinase. In: Hagedorn, C.H., Rice, C.M. (eds) The Hepatitis C Viruses. Current Topics in Microbiology and Immunology, vol 242. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-59605-6_8

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  • DOI: https://doi.org/10.1007/978-3-642-59605-6_8

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