Abstract

Hepatitis C virus (HCV) causes a persistent infection, chronic hepatitis, and leads to hepatocellular carcinoma. Nonstructural protein 3 (NS3) of HCV possesses protease, nucleoside triphosphatase, and helicase activities. Using the yeast two hybrid assay, we identified Sm-D1, a host protein that binds to NS3. Sm-D1 is a component of small nuclear ribonucleoprotein (snRNP) complexes which are associated with autoimmune disease. Sm-D1 has Gly-Arg (GR) repeats at the C terminus, which contains dimethylarginine modified by post-translational modification and may constitute an immunoreactive determinant. Deletion mutants revealed that the C-terminal region of Sm-D1 containing the GR repeats was the binding region for NS3, and the expression feature of Sm-D1 was affected by co-expression of NS3. Immunostaining assay demonstrated that NS3 was also present in the nucleus of cells overexpressing Sm-D1, although it was usually found in cytoplasm. The localization of NS3 could change following interaction with Sm-D1 and affect the function of Sm-D1.

References

  • 1Abramovich, C., Yakobson, B., Chebath, J., and Revel, M. 1997. A protein-arginine methyltransferase binds to the intracytoplasmic domain of the IFNAR1 chain in the type I interferon receptor. EMBO J. 16: 260–266.ViewCASPubMedWeb of Science®Google Scholar
  • 2Achsel, T., Stark, H., and Luhrmann, R. 2001. The Sm domain is an ancient RNA-binding motif with oligo (U) specificity. Proc. Natl. Acad. Sci. U.S.A. 98: 3685–3689.ViewCASPubMedWeb of Science®Google Scholar
  • 3Banerjee, R., and Dasgupta, A. 2001. Specific interaction of hepatitis C virus protease/helicase NS3 with the 3′-terminal sequences of viral positive- and negative-strand RNA. J. Virol. 75: 1708–1721.ViewCASPubMedWeb of Science®Google Scholar
  • 4Bartenschlager, R., Ahlborn Laake, L., Mous, J., and 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.ViewCASPubMedWeb of Science®Google Scholar
  • 5Bartenschlager, R., and Lohmann, V. 2000. Replication of hepatitis C virus. J. Gen. Virol. 81: 1631–1648.ViewCASPubMedWeb of Science®Google Scholar
  • 6Blanton, S., Srinivasan, A., and Rymond, B.C. 1992. PRP38 encodes a yeast protein required for pre-mRNA splicing and maintenance of stable U6 small nuclear RNA levels. Mol. Cell. Biol. 12: 3939–3947.ViewCASPubMedWeb of Science®Google Scholar
  • 7Bordonne, R. 2000. Functional characterization of nuclear localization signals in yeast Sm proteins. Mol. Cell. Biol. 20: 7943–7954.ViewCASPubMedWeb of Science®Google Scholar
  • 8Brahms, H., Raymackers, J., Union, A., De Keyser, F., Meheus, L., and Luhrmann, R. 2000. The C-terminal RG dipeptide repeats of the spliceosomal Sm proteins D1 and D3 contain symmetrical dimethylarginines, which form a major B-cell epitope for anti-Sm autoantibodies. J. Biol. Chem. 275: 17122–17129.ViewCASPubMedWeb of Science®Google Scholar
  • 9 C.B. Burge, T. Tuschl, and P.A. Sharp (eds), 1999. In The RNA world, 2nd ed, p. 525–560.Google Scholar
  • 10Choo, Q.L., Kuo, G., Weiner, A.J., Overby, L.R., Bradley, D.W., and Houghton, M. 1989. Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science 244: 359–362.ViewCASPubMedWeb of Science®Google Scholar
  • 11Fisher, D.E., Conner, G.E., Reeves, W.H., Wisniewolski, R., and Blobel, G. 1985. Small nuclear ribonucleoprotein particle assembly in vivo: demonstration of a 6S RNA-free core precursor and posttranslational modification. Cell 42: 751–758.ViewCASPubMedWeb of Science®Google Scholar
  • 12Gary, J.D., and Clarke, S. 1998. RNA and protein interactions modulated by protein arginine methylation. Prog. Nucleic Acid Res. Mol. Biol. 61: 65–131.ViewCASPubMedWeb of Science®Google Scholar
  • 13Gary, J.D., Lin, W.J., Yang, M.C., Herschman, H.R., and Clarke, S. 1996. The predominant protein-arginine methyltransferase from Saccharomyces cerevisiae. J. Biol. Chem. 271: 12585–12594.ViewCASPubMedWeb of Science®Google Scholar
  • 14Gerke, V., and Steitz, J.A. 1986. A protein associated with small nuclear ribonucleoprotein particles recognizes the 3′ splice site of premessenger RNA. Cell 47: 973–984.ViewCASPubMedWeb of Science®Google Scholar
  • 15Hoofnagle, J.H. 1997. Hepatitis C: the clinical spectrum of disease. Hepatology 26: 15S–20S.ViewCASPubMedWeb of Science®Google Scholar
  • 16Ishido, S., Muramatsu, S., Fujita, T., Iwanaga, Y., Tong, W.Y., Katayama, Y., Itoh, M., and Hotta, H. 1997. Wild-type, but not mutant-type, p53 enhances nuclear accumulation of the NS3 protein of hepatitis C virus. Biochem. Biophys. Res. Commun. 230: 431–436.ViewCASPubMedWeb of Science®Google Scholar
  • 17James, J.A., Mamula, M.J., and Harley, J.B. 1994. Sequential autoantigenic determinants of the small nuclear ribonucleoprotein Sm-D1 shared by human lupus autoantibodies and MRL lpr/lpr antibodies. Clin. Exp. Immunol. 98: 419–426.ViewCASPubMedWeb of Science®Google Scholar
  • 18Kim, D.W., Gwack, Y., Han, J.H., and Choe, J. 1995. C-terminal domain of the hepatitis C virus NS3 protein contains an RNA helicase activity. Biochem. Biophys. Res. Commun. 215: 160–166.ViewCASPubMedWeb of Science®Google Scholar
  • 19Kuo, G., Choo, Q.L., Alter, H.J., Gitnick, G.L., Redeker, A.G., Purcell, R.H., Miyamura, T., Dienstag, J.L., Alter, M.J., Stevens, C.E., Tegtmeier, G.E., Bonino, F., Colombo, M., Lee, W.S., Kuo, C., Berger, K., Shuster, J.R., Overby, L.R., Bradley, D.W., and Houghton, M. 1989. An assay for circulating antibodies to a major etiologic virus of human non-A, non-B hepatitis. Science 244: 362–364.ViewCASPubMedWeb of Science®Google Scholar
  • 20Kwun, H.J., Jung, E.Y., Ahn, J.Y., Lee, M.N., and Jang, K.L. 2001. p53-dependent transcriptional repression of p21(waf1) by hepatitis C virus NS3. J. Gen. Virol. 82: 2235–2241.ViewCASPubMedWeb of Science®Google Scholar
  • 21Liautard, J.P., Sri Widada, J., Brunel, C., and Jeanteur, P. 1982. Structural organization of ribonucleoproteins containing small nuclear RNAs from HeLa cells. Proteins interact closely with a similar structural domain of U1, U2, U4 and U5 small nuclear RNAs. J. Mol. Biol. 162: 623–643.ViewCASPubMedWeb of Science®Google Scholar
  • 22Lin, W.J., Gary, J.D., Yang, M.C., Clarke, S., and Herschman, H.R. 1996. The mammalian immediate-early TIS21 protein and the leukemia-associated BTG1 protein interact with a protein-arginine N-methyltransferase. J. Biol. Chem. 271: 15034–15044.ViewCASPubMedWeb of Science®Google Scholar
  • 23Meerovitch, K., Pelletier, J., and Sonenberg, N. 1989. A cellular protein that binds to the 5′-noncoding region of poliovirus RNA: implications for internal translation initiation. Genes Dev. 3: 1026–1034.ViewCASPubMedWeb of Science®Google Scholar
  • 24Mitsuda, T., Eisenberg, R.A., and Cohen, P.L. 1992. The murine Sm-D autoantigen: multiple genes, genetic polymorphism, evolutionary conservation and lack of intervening sequences in the coding region. J. Autoimmun. 5: 277–287.ViewCASPubMedWeb of Science®Google Scholar
  • 25Muramatsu, S., Ishido, S., Fujita, T., Itoh, M., and Hotta, H. 1997. Nuclear localization of the NS3 protein of hepatitis C virus and factors affecting the localization. J. Virol. 71: 4954–4961.ViewCASPubMedWeb of Science®Google Scholar
  • 26Ou, Y., Sun, D., Sharp, G.C., and Hoch, S.O. 1997. Screening of SLE sera using purified recombinant Sm-D1 protein from a baculovirus expression system. Clin. Immunol. Immunopathol. 83: 310–317.ViewCASPubMedWeb of Science®Google Scholar
  • 27Reed, K.E., and Rice, C.M. 2000. Overview of hepatitis C virus genome structure, polyprotein processing, and protein properties. Curr. Top. Microbiol. Immunol. 242: 55–84.ViewCASPubMedWeb of Science®Google Scholar
  • 28Rokeach, L.A., Haselby, J.A., and Hoch, S.O. 1992. Overproduction of a human snRNP-associated Sm-D autoantigen in Escherichia coli and Saccharomyces cerevisiae. Gene 118: 247–253.ViewCASPubMedWeb of Science®Google Scholar
  • 29Rokeach, L.A., Haselby, J.A., and Hoch, S.O. 1988. Molecular cloning of a cDNA encoding the human Sm-D autoantigen. Proc. Natl. Acad. Sci. U.S.A. 85: 4832–4836.ViewCASPubMedWeb of Science®Google Scholar
  • 30Rokeach, L.A., Jannatipour, M., Haselby, J.A., and Hoch, S.O. 1992. Mapping of the immunoreactive domains of a small nuclear ribonucleoprotein-associated Sm-D1 autoantigen. Clin. Immunol. Immunopathol. 65: 315–324.ViewCASPubMedWeb of Science®Google Scholar
  • 31Sakamuro, D., Furukawa, T., and Takegami, T. 1995. Hepatitis C virus nonstructural protein NS3 transforms NIH 3T3 cells. J. Virol. 69: 3893–3896.ViewCASPubMedWeb of Science®Google Scholar
  • 32 J. Sambrook, E.F. Fritsch, and T. Maniatis (eds), 1989. Molecular cloning: a laboratory manual 2nd ed, p. 17.48.Google Scholar
  • 33Sauterer, R.A., Goyal, A., and Zieve, G.W. 1990. Cytoplasmic assembly of small nuclear ribonucleoprotein particles from 6 S and 20 S RNA-free intermediates in L929 mouse fibroblasts. J. Biol. Chem. 265: 1048–1058.CASPubMedWeb of Science®Google Scholar
  • 34Tan, E.M. 1989. Antinuclear antibodies: diagnostic markers for autoimmune diseases and probes for cell biology. Adv. Immunol. 44: 93–151.ViewCASPubMedWeb of Science®Google Scholar
  • 35Tazi, J., Alibert, C., Temsamani, J., Reveillaud, I., Cathala, G., Brunel, C., and Jeanteur, P. 1986. A protein that specifically recognizes the 3′ splice site of mammalian pre-mRNA introns is associated with a small nuclear ribonucleoprotein. Cell 47: 755–766.ViewCASPubMedWeb of Science®Google Scholar
  • 36Van Mühlen, C.A., and Tan, E.M. 1995. Autoantibodies in the diagnosis of systemic rheumatic diseases. Semin. Arthritis. Rheum. 24: 323–358.ViewPubMedWeb of Science®Google Scholar
  • 37Wagatsuma, M., Asami, N., Miyachi, J., Uchida, S., Watanabe, H., and Amann, E. 1993. Antibody recognition of the recombinant human nuclear antigens RNP 70 kD, SS-A, SS-B, Sm-B, and Sm-D by autoimmune sera. Mol. Immunol. 30: 1491–1498.ViewCASPubMedWeb of Science®Google Scholar
  • 38Wolk, B., Sansonno, D., Krausslich, H.G., Dammacco, F., Rice, C.M., Blum, H.E., and Moradpour, D. 2000. Subcellular localization, stability, and trans-cleavage competence of the hepatitis C virus NS3-NS4A complex expressed in tetracycline-regulated cell lines. J. Virol. 74: 2293–2304.ViewCASPubMedWeb of Science®Google Scholar
  • 39Wright Minogue, J., Yao, N., Zhang, R., Butkiewicz, N.J., Baroudy, B.M., Lau, J.Y., and Hong, Z. 2000. Cross-genotypic interaction between hepatitis C virus NS3 protease domains and NS4A cofactors. J. Hepatol. 32: 497–504.ViewCASPubMedWeb of Science®Google Scholar
  • 40Zhang, D., Abovich, N., and Rosbash, M. 2001. A biochemical function for the Sm complex. Mol. Cell. 7: 319–329.ViewCASPubMedWeb of Science®Google Scholar

References

https://doi.org/10.1111/j.1348-0421.2003.tb03423.x