1887

Abstract

The picornavirus membrane-associated polypeptide 2C is believed to be required for viral RNA synthesis. Hepatitis A virus (HAV)- and human rhinovirus (HRV)-encoded recombinant 2C proteins have been expressed, purified and examined for their ability to interact with the terminal sequences of viral positive- and negative-strand RNAs. The results demonstrate that both the HAV- and the HRV-encoded 2C polypeptide specifically interact with the 3′-terminal sequences of the negative-strand RNA, but not with the complementary sequences at the 5′ terminus of the positive-strand RNA. This interaction was detected by both mobility gel shift and UV cross-linking assays. Furthermore, complex formation exhibited dose-dependency and competition assays confirmed specificity. These results are consistent with our previous observation using the poliovirus 2C protein. The implication of the picornavirus 2C protein binding to the 3′-terminal sequence of the negative-strand untranslated region in viral RNA synthesis is discussed.

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2001-11-01
2024-04-27
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References

  1. Aldabe R., Carrasco L. 1995; Induction of membrane proliferation by poliovirus proteins 2C and 2BC. Biochemical and Biophysical Research Communications 206:64–76
    [Google Scholar]
  2. Andino R., Rieckhof G. E., Baltimore D. 1990; A functional ribonucleoprotein complex forms around the 5′ end of poliovirus RNA. Cell 63:369–380
    [Google Scholar]
  3. Andino R., Rieckhof G. E., Achacoso P. L., Baltimore D. 1993; Poliovirus RNA synthesis utilizes an RNP complex formed around the 5′-end of viral RNA. EMBO Journal 12:3587–3598
    [Google Scholar]
  4. Banerjee R., Dasgupta A. 2001; Specific interaction of hepatitis C virus protease/helicase NS3 with the 3′-terminal sequences of the viral positive- and negative-strand RNA. Journal of Virology 75:1708–1721
    [Google Scholar]
  5. Banerjee R., Echeverri A., Dasgupta A. 1997; Poliovirus-encoded 2C polypeptide specifically binds to the 3′-terminal sequences of viral negative-strand RNA. Journal of Virology 71:9570–9578
    [Google Scholar]
  6. Banerjee R., Igo M., Izumi R., Datta U., Dasgupta A. 2000; In vitro replication of RNA viruses. In RNA Viruses: A Practical Approach pp 158–164 Edited by Cann A. J. New York: Oxford University Press;
    [Google Scholar]
  7. Banerjee R., Tsai W., Kim W., Dasgupta A. 2001; Interaction of poliovirus-encoded 2C/2BC polypeptide with the 3′ terminus negative-strand cloverleaf requires an intact stem–loop b. Virology 280:41–51
    [Google Scholar]
  8. Bienz K., Egger D., Pasamontes L. 1987; Association of polioviral proteins of the P2 genomic region with the viral replication complex and virus-induced membrane synthesis as visualized by electron microscopic immunocytochemistry and autoradiography. Virology 160:220–226
    [Google Scholar]
  9. Bienz K., Egger D., Troxler M., Pasamontes L. 1990; Structural organization of poliovirus RNA replication is mediated by viral proteins of the P2 genomic region. Journal of Virology 64:1156–1163
    [Google Scholar]
  10. Bienz K., Egger D., Pfister T., Troxler M. 1992; Structural and functional characterization of the poliovirus replication complex. Journal of Virology 66:2740–2747
    [Google Scholar]
  11. Brown E. A., Day S. P., Jansen R. W., Lemon S. M. 1991; The 5′ nontranslated region of hepatitis A virus RNA: secondary structure and elements required for translation in vitro. Journal of Virology 65:5828–5838
    [Google Scholar]
  12. Cho M. W., Teterina N. L., Egger D., Bienz K., Ehrenfeld E. 1994; Membrane rearrangement and vesicle induction by recombinant poliovirus 2C and 2BC in human cells. Virology 202:129–145
    [Google Scholar]
  13. Cohen J. I., Ticehurst J. R., Purcell R. H., Buckler-White A., Baroudy B. M. 1987; Complete nucleotide sequence of wild-type hepatitis A virus: comparison with different strains of hepatitis A virus and other picornaviruses. Journal of Virology 61:50–59
    [Google Scholar]
  14. Echeverri A. C., Dasgupta A. 1995; Amino-terminal region of poliovirus 2C protein mediates membrane binding. Virology 208:540–553
    [Google Scholar]
  15. Echeverri A. C., Banerjee R., Dasgupta A. 1998; Amino terminal region of poliovirus 2C is sufficient for membrane binding. Virus Research 54:217–223
    [Google Scholar]
  16. Gorbalenya A. F., Blinov V. M., Donchenko A. P., Koonin E. V. 1989; An NTP-binding motif is the most conserved sequence in a highly diverged monophyletic group of proteins involved in positive strand RNA viral replication. Journal of Molecular Evolution 28:256–268
    [Google Scholar]
  17. Harris K. S., Xiang W., Alexander L., Lane W. S., Paul A. V., Wimmer E. 1994; Interaction of poliovirus polypeptide 3CDpro with the 5′ and 3′ termini of the poliovirus genome. Identification of viral and cellular cofactors needed for efficient binding. Journal of Biological Chemistry 269:27004–27014
    [Google Scholar]
  18. Kitamura N., Semler B. L., Rothberg P. G., Larsen G. R., Adler C. J., Dorner J. A., Emini E. A., Hanecak R., Lee J. J., van der Werf S., Anderson C. W., Wimmer E. 1981; Primary structure, gene organization and polypeptide expression of poliovirus RNA. Nature 291:547–553
    [Google Scholar]
  19. Klein M., Eggers H. J., Nelsen-Salz B. 2000; Echovirus-9 protein 2C binds single-stranded RNA unspecifically. Journal of General Virology 81:2481–2484
    [Google Scholar]
  20. Kusov Y. Y., Gauss-Muller V. 1997; In vitro RNA binding of the hepatitis A virus proteinase 3C (HAV 3Cpro) to secondary structure elements within the 5′ terminus of the HAV genome. RNA 3:291–302
    [Google Scholar]
  21. Kusov Y. Y., Morace G., Probst C., Gauss-Muller V. 1997; Interaction of hepatitis A virus (HAV) precursor proteins 3AB and 3ABC with the 5′ and 3′ termini of the HAV RNA. Virus Research 51:151–157
    [Google Scholar]
  22. Kusov Y. Y., Probst C., Jecht M., Jost P. D., Gauss-Muller V. 1998; Membrane association and RNA binding of recombinant hepatitis A virus protein 2C. Archives of Virology 143:931–944
    [Google Scholar]
  23. Lemon S. M., Robertson B. H. 1993; Current perspectives in the virology and molecular biology of hepatitis A virus. Seminars in Virology 4:285–295
    [Google Scholar]
  24. Mellits K. H., Meredith J. M., Rohll J. B., Evans D. J., Almond J. W. 1998; Binding of a cellular factor to the 3′ untranslated region of the RNA genomes of entero- and rhinoviruses plays a role in virus replication. Journal of General Virology 79:1715–1723
    [Google Scholar]
  25. Mirzayan C., Wimmer E. 1994; Biochemical studies on poliovirus polypeptide 2C: evidence for ATPase activity. Virology 189:176–187
    [Google Scholar]
  26. Monika J., Probst C., Gauss-Muller V. 1988; Membrane permeability induced by HAV proteins 2B and 2BC and proteolytic processing of HAV 2BC. Virology 252:218–227
    [Google Scholar]
  27. Parsley T. B., Towner J. S., Blyn L. B., Ehrenfeld E., Semler B. L. 1997; Poly(rC) binding protein 2 forms a ternary complex with the 5′-terminal sequences of poliovirus RNA and the viral 3CD proteinase. RNA 3:1124–1134
    [Google Scholar]
  28. Pfister T., Wimmer E. 1999; Characterization of the nucleoside triphosphatase activity of poliovirus protein 2C reveals a mechanism by which guanidine inhibits poliovirus replication. Journal of Biological Chemistry 274:6992–7001
    [Google Scholar]
  29. Rodriguez P. L., Carrasco L. 1993; Poliovirus protein 2C has ATPase and GTPase activities. Journal of Biological Chemistry 268:8105–8110
    [Google Scholar]
  30. Rodriguez P. L., Carrasco L. 1995; Poliovirus protein 2C contains two regions involved in RNA binding activity. Journal of Biological Chemistry 270:10105–10112
    [Google Scholar]
  31. Rohll J. B., Percy N., Ley R., Evans D. J., Almond J. W., Barclay W. S. 1994; The 5′-untranslated region of picornavirus RNAs contain independent functional domains essential for RNA replication and translation. Journal of Virology 68:4384–4391
    [Google Scholar]
  32. Stanway G., Hughes P. J., Mountford C. R., Minor P. D., Almond J. W. 1984; The complete nucleotide sequence of a common cold virus: human rhinovirus 14. Nucleic Acid Research 12:7859–7875
    [Google Scholar]
  33. Teterina N. L., Kean K. M., Gorbalenya A. E., Agol V. I., Girard M. 1992; Analysis of the functional significance of amino acid residues in the putative NTP-binding pattern of the poliovirus 2C protein. Journal of General Virology 73:1977–1980
    [Google Scholar]
  34. Teterina N. L., Bienz K., Egger D., Gorbalenya A. G., Ehrenfeld E. 1997; Induction of intracellular membrane rearrangements by HAV proteins 2C and 2BC. Virology 237:66–77
    [Google Scholar]
  35. Todd S., Nguyen J. H., Semler B. L. 1995; RNA–protein interactions directed by the 3′ end of the human rhinovirus genomic RNA. Journal of Virology 69:3605–3614
    [Google Scholar]
  36. Xiang W., Harris K. S., Alexander L., Wimmer E. 1995; Interaction between the 5′-terminal cloverleaf and 3AB/3CDpro of poliovirus is essential for RNA replication. Journal of Virology 69:3658–3667
    [Google Scholar]
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