An astrovirus frameshift signal induces ribosomal frameshifting in vitro


Expression of the astrovirus RNA-dependent RNA polymerase has been hypothesized to be regulated by (− 1) ribosomal frameshifting. Sequence analysis of the 70 nucleotide region between open reading frames 1a and 1b indicates the presence of a shifty heptamer consensus sequence and downstream sequences that may be needed for ribosomal frameshifting. We constructed four astrovirus cassettes that spanned this region and inserted each into the rhesus rotavirus VP4 gene. The constructs were expressed in an in vitro system, and products were immunoprecipitated by rotavirus amino and carboxy terminal-specific monoclonal antibodies. Ribosomal frameshifting, at an efficiency of 6–7%, was demonstrated in all constructs containing the shifty heptamer and stem-loop. Deletion of the downstream sequence potentially involved in pseudoknot formation did not affect frameshifting efficiency. However, deletion of the shifty heptamer resulted in no detectable frameshift activity.


  1. 1.

    Belcourt MF, Farabaugh PJ (1990) Ribosomal frameshifting in the yeast retrotransposon Ty: tRNAs induce slippage on a 7 nucleotide minimal site. Cell 62: 339–352

  2. 2.

    Bredenbeek PJ, Pachuk CJ, Noten AFH, Charite J, Luytjes W, Weiss SR, Spaan WJM (1990) The primary structure and expression of the second open reading frame of the polymerase gene of the coronavirus MHV-A59; a highly conserved polymerase is expressed by an efficient ribosomal frameshifting mechanism. Nucleic Acids Res 18: 1825–1832

  3. 3.

    Brierley I (1993) Probing the mechanism of ribosomal frameshifting on viral RNAs. Biochem Soc Trans 21: 822–826

  4. 4.

    Brierley I, Bournsnell MEG, Binns MM, Bilimoria B, Blok VC, Brown TDK, Inglis SC, (1987) An efficient ribosomal frame-shifting signal in the polymerase-encoding region of the coronavirus avian infectious bronchitis virus. EMBO J. 6: 3779–3785

  5. 5.

    Cox GJ, Matsui SM, Lo RS, Hinds M, Bowden RA, Hackman RC, Meyer WG, Mori M, Tarr PI, Oshiro LS, Ludert JE, Meyers JD, McDonald GB (1994) Etiology and outcome of diarrhoea after marrow transplantation: a prospective study. Gastroenterology 107: 1398–1407

  6. 6.

    Cruz JR, Bartlett JE, Herrmann JE, Caceres P, Blacklow NR, Cano F, (1992) Astrovirus-associated diarrhoea among Guatemalan ambulatory rural children. J Clin Microbiol 30: 1140–1144

  7. 7.

    Dinman JD, Icho T, Wickner RB (1991) A -1 ribosomal frameshift in a double-stranded RNA virus of yeast forms a gag-pol fusion protein. Proc Natl Acad Sci USA 88: 174–178

  8. 8.

    Falk H, Mador N, Udi R, Panet A, Honigman A (1993) Two cis- acting signals control ribosomal frameshift between human T-cell leukemia virus type II gag and pro genes. J Virol 67: 6273–6277

  9. 9.

    Grohmann GS, Glass RI, Pereira HG, Monroe SS, Hightower AW, Weber R, Bryan RT (1993) Enteric viruses and diarrhoea in HIV-infected patients. N Engl J Med 329: 14–20

  10. 10.

    Hatfield DL, Levin JG, Rein A, Oroszlan S (1992) Translational suppression in retroviral gene expression. Adv Virus Res 41: 193–239

  11. 11.

    Hatfield DL, Smith DWE, Lee BJ, Worland PJ, Oroszlan S (1990) Structure and function of suppressor tRNAs in higher eukaryotes. Crit Rev Biochem Mol Biol 25: 71–96

  12. 12.

    Herrmann JE, Taylor DN, Echeverria P, Blacklow NR (1991) Astroviruses as a cause of gastroenteritis in children. N Engl J Med 324: 1157–1760

  13. 13.

    Jiang B, Monroe SS, Koonin EV, Stine SE, Glass RI (1993) RNA sequence of astrovirus: distinctive genomic organization and a putative retrovirus-like ribosomal frameshifting signal that directs the viral replicase synthesis. Proc Natl Acad Sci USA 90: 10539–10543

  14. 14.

    Jacks T (1990) Translational suppression in gene expression in retroviruses and retrotransposons. Curr Top Microbiol Immunol 157: 93–124

  15. 15.

    Jacks T, Madhani HD, Masiarz FR, Varmus HE (1988) Signals for ribosomal frameshifting in the rous sarcoma virus. Cell 55: 447–458

  16. 16.

    Jacks T, Varmus HE (1985) Expression of the rous sarcoma virus pol gene by ribosomal frameshifting. Science 230: 1237–1242

  17. 17.

    Kozak M (1987) An analysis of 5′-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res 15: 8125–8148

  18. 18.

    Kurtz JB, Lee TW (1984) Human astrovirus serotypes. Lancet ii: 1405

  19. 19.

    Lee TW, Kurtz JB (1981) Serial propagation of astrovirus in tissue culture with the aid of trypsin. J Gen Virol 57: 421–424

  20. 20.

    Lee TW, Kurtz JB (1994) Prevalence of human astrovirus serotypes in the Oxford region 1976–92, with evidence for two new serotypes. Epidemiol Infect 112: 187–193

  21. 21.

    Lewis TL, Greenberg HB, Herrmann JE, Smith LS, Matsui SM (1994) Analysis of astrovirus serotype 1 RNA, identification of the viral RNA-dependent RNA polymerase motif, and expression of a viral structural protein. J Virol 68: 77–83

  22. 22.

    Mackow ER, Shaw RD, Matsui SM, Vo PT, Dang MN, Greenberg HB (1988) The rhesus rotavirus gene encoding protein VP3: location of amino acids involved in homologous and heterologus rotavirus neutralization and identification of a putative fusion region. Proc Natl Acad Sci USA 85: 645–649

  23. 23.

    Mackow ER, Yamanaka MY, Dang MN, Greenberg HB (1990) DNA amplification-restricted transcription-translation: rapid analysis of rhesus rotavirus neutralization sites. Proc Natl Acad Sci USA 87: 518–522

  24. 24.

    Madeley CR, Cosgrove BP (1975) Viruses in infantile gastroenteritis. Lancet ii: 124

  25. 25.

    Marczinke B, Bloys AJ, Brown TDK, Willcocks MM, Carter MJ, Brierley I (1994) The human astrovirus RNA-dependent RNA polymerase coding region is expressed by ribosomal frameshifting. J Virol 68: 5588–5595

  26. 26.

    Matsui SM, Kim JP, Greenberg HB, Young LM, Smith LS, Lewis TL, Herrmann JE, Blacklow NR, Dupuis K, Reyes GR (1993) Cloning and characterization of human astrovirus immunoreactive epitopes. J Virol 67: 1712–1715

  27. 27.

    Monroe SS, Jiang B, Stine SE, Koopmans M, Glass RI (1993) Subgenomic RNA sequence of human astrovirus supports classification ofAstroviridae as a new family of RNA viruses. J Virol 67: 3611–3614

  28. 28.

    Monroe SS, Stine SE, Gorelkin L, Herrmann JE, Blacklow NR, Glass RI (1991) Temporal synthesis of proteins and RNAs during human astrovirus infection of cultured cells. J Virol 65: 641–648

  29. 29.

    Reil H, Kollmus H, Weidle UH, Hauser HJ (1993) A heptanucleotide sequence mediates ribosomal frameshifting in mammalian cells. J Virol 67: 5579–5584

  30. 30.

    Shaw RD, Vo PT, Offit PA, Coulson BS, Greenberg HB (1986) Antigenic mapping of the surface proteins rhesus rotavirus. Virology 155: 434–451

  31. 31.

    Willcocks MM, Brown TDK, Madeley CR, Carter MJ (1994) The complete sequence of a human astrovirus. J Gen Virol 75: 1785–1788

  32. 32.

    Willcocks MM, Carter MJ (1993) Identification and sequence determination of the capsid protein of human astrovirus serotype 1. FEMS Microbiol Lett 114: 1–8

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Lewis, T.L., Matsui, S.M. An astrovirus frameshift signal induces ribosomal frameshifting in vitro. Archives of Virology 140, 1127–1135 (1995).

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  • Nucleotide
  • Monoclonal Antibody
  • Sequence Analysis
  • Consensus Sequence
  • Carboxy