Abstract
Although studies of picornavirus proteins and their biosynthesis in vivo have been conducted with approximately equal intensity for both poliovirus and encephalomyocarditis (EMC) virus (l), the majority of in vitro translation studies have been performed using the mouse virus RNA. Beginning in the early 1970’s, several different laboratories reported the synthesis of virus-specific proteins in preincubated cell-free systems derived from Krebs II, Ehrlich or mouse plasmacytoma ascites cells, or from cultured L cells (2–13). In general, a variety of different sized polypeptides, including those with molecular weights over 100,000 daltons, were produced, but, unlike the various viral proteins synthesized in vivo, the in vitro products appear to arise from premature termination of translation rather than from proteolytic cleavage of a complete translate. Complete translation of the entire viral RNA molecule may occur in rare instances (5), but most of the polypeptides synthesized in vitro contain amino acid sequences derived from a common amino terminus, and the majority of the products contain the tryptic (or cyanogen bromide) peptide fragments which are present in the virus capsid proteins. These proteins are known to be encoded by nucleotide sequences within the 5′ half of the viral RNA (l). A diagram of the overlap in amino sequences of the major translation products (I-VI) of EMC virus RNA by a Krebs or Ehrlich ascites cell-free system is shown below. The data are from Hunt (13).
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EUEGKEET, R.R. On the structxire and morphogenesis of picornaviruses. In Comprehensive Tirology. Fraenkel-Conrat, H. and Wagner, R.R. eds., (1976), Vol.6, pp.131–213, Plenum Press, New York.
KEEE, I.M. and MAETIN, E.M. Virus protein synthesis in animal cell-free systems: Nature of the products synthesized in response to MA of EMC virus. J. Virol. (1971), 7, 458–447
DOBOS, P., KERR, I.M. and MARTIN, E.M. Synthesis of capsid and non-capsid viral proteins in response to EMG virus MA in animal cell-free systems. J. Virol. (1971), 8, 491–499.
KERR, I.M., BEOWN, R.M. and TOYELL, D.R. Gharacterization of the polypeptides formed in response to EMC virus MA in a cell-free system from mouse ascites tumor cells. J. Virol. (1972), 10, 73–81.
ESTEBAN, M. and KERR, I.M. The synthesis of EMC virus polypeptides in infected L-cells and cell-free systems. Eur. J. Biochem. (1974), 45, 567–576.
MATHEWS, M.B. and KORNER, A. Mammalian cell-free protein synthesis directed by viral RNA. Eur. J. Biochem. (1970), 17, 328–338.
SMITH, A.E., MARGKER, K.A. and MATHEWS, M.B. Translation of RNA from EMC virus in a maimnalian cell-free system. Nature (London) (1970), 225, 184–187.
MATHEWS, M.B. and OSBORN, M. The rate of polypeptide chain elongation in a cell-free system from Krebs II ascites cells. Biochem. Biophys. Acta. (1974), 340 147–152.
BOIME, I., AVIV, H. and LEBER, P. Protein synthesis directed by EMC virus RNA. II. The in vitro synthesis of high molecular weight proteins and elements of the viral capsid. Biochem. Biophys. Res. Comm. (1971), 45, 788–795.
BOIME, I. and LEBER, P. Protein synthesis directed by EMC virus RNA. III.Biscrete polypeptides translated from a monocistronic messenger in vitro. Arch. Biochem. Biophys. (1972), 153, 706–713.
EGGEN, K.L. and SHATKIN, A.J. In vitro translation of cardiovirus ribonucleic acid by mammalian cell-free extracts. J. Virol. (1972), 9, 636–645.
LAWRENCE, C. and THACH, R.E. Encephalomyocarditis virus infection of mouse plasmacytoma cells. I. Inhibition of cellular protein synthesis. J. Virol. (1974), 14, 598–610.
HUNT, L.A. In vitro translation of EMC viral RNA: Synthesis of capsid precursor-like polypeptides. Virology (1976), 70, 484–492.
HUNT, L.A. Mechanism of premature polypeptide termination in a mouse ascites cell-free system programmed by EMC RNA. J. Virol. (1976), 18, 788–792.
HACKETT, P.B., EGBERTS, E. and TRAUB, P. Translation of ascites and mengovirus RNA in fractionated cell-free systems from iminfected and mengovirus-infected Ehrlich ascites tumor cells. Eur. J. Biochem. (1978), 83, 341–352.
LAWRENCE, C. and THACH, R.E. Identification of a viral protein involved in post-translational maturation of the EMC virus capsid precursor. J. Virol. (1975), 15, 918–928.
PELHAM, H.R.B. Translation of EMC virus RNA in vitro yields an active proteolytic processing enzyme. Eur. J. Biochem. (1978), 85, 457–462.
PELHAM, H.R.B. and JACKSON, R.J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur. J. Biochem. (1978), 85, 457–462.
SYITKIN, Y.Y., GINEVSKAYA, V.A., UGAROVA, T.Y. and AGOL, V.I. A cell-free model of the EMC virus-induced inhibition of host cell protein synthesis. Virology (1978), 87, 199–205.
LEYINTOW, L. The reproduction of picornaviruses. In Comprehensive Virology. Praenkel-Conrat, H. and Wagner, R.R. eds. (1974), Vol.2, pp.109–169, Plenum Press, New York.
ROBERTS, B.E. and PATERSON, B.M. Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S in a cell-free system from commercial wheat germ. Proc. Nat. Acad. Sci. U.S.A. (1973), 70. 2330–2334.
EHRENFELD, E. and LUND, H. Untranslated vesicular stomatitis virus messenger RNA after poliovirus infection. Virology (1977), 80, 279–308.
LODISH, H.F. and ROSE, J.K. Relative importance of 7-methyl guanosine in ribosome binding and translation of VSV mRNA in wheat germ and reticulocyte cell-free systems. J. Biol. Ghem., (1977), 252, 1181–1188.
LEE, Y.F., NOMOTO, A., DETJEN, B.M. and WIMMER, E. A protein covalently linked to poliovirus genome RNA. Proc. Nat. Acad. Sci. U.S.A. (1977), 74, 59–65.
FLANEGAN, J.B., PETTERSSON, R.F., AMBROS, V., HEWLETT, M. and BALTIMORE, D. Covalent linkage of a protein to a defined nucleotide sequence at the 5’ terminus of virion and replicative intermediate RNA of poliovirus. Proc. Nat. Acad. Sci. U.S.A., 74, 961–965.
VILLA-KOMAROPP, L., GUTTMAN, N., BALTIMORE, D. and LODISH, H.P. Complete translation of poliovirus RNA in a exikaryotic cell-free system. Proc. Nat. Acad. Sci. U.S.A. (1975), 72, 4157–4161.
ROSE, J.K., TRASCHEL, H., LEONG, K. and BALTIMORE, D. Inhibition of translation by poliovirus: Inactivation of a specific initiation factor. Proc. Nat. Acad. Sci. U.S.A. (1978), 75, 2732–2736.
SHAPRITZ, D.A., PADILLA, M., GANAANI, D., GRONER, Y., WEINSTEIN, J.A., BAR-JOSEPH, M. and MERRICK, W.C. Initiation factor eIP-4B-dependent recognition and translation of capped versus uncapped eukaryotic mRNA. J. Biol. Chem. (1978) in press.
PERNANDE-ZMDNOZ, R. and DARNELL, J.E. Structural difference between the 5’ terminus of viral and cellular mRNA in poliovirus-infected cells: Possible basis for the inhibition of host protein synthesis. J. Virol. (1976), 18, 719–726.
HEWLETT, M.J., ROSE, J.K. and BALTIMORE, D. 5’ terminal structure of poliovirus polyribosomal RNA is pUp. Proc. Nat. Acad. Sci. U.S.A. (1976), 73, 327–330.
NOMOTO, A., LEE, Y.F. and WIMMER, E. The 5’ end of poliovirus mRNA is not capped with m7G(5’)ppp(5’)Np. Proc. Nat. Acad. Sci. U.S.A. (1976), 73, 375–380.
CELMA, M.L. and EHRENFELD, E. Effect of poliovirus double-stranded RNA on viral and host cell protein synthesis. Proc. Nat. Acad. Sci. U.S.A. (1974), 74, 2440–2444.
KAUPMACT, Y., GOLDSTEIN, E. and PENMAN, S. Poliovirus-induced inhibition of polsrpeptide initiation to vitro on native polyribosomes. Proc. Nat. Acad. Sci. U.S.A. (1976), 73, 1834–1838.
HELENTJARIS, T. and EHRENPELD, E. Control of protein synthesis in extracts from poliovirus-infected cells. J. Virol. (1978), 26, 510–521.
HACKETT, P.B., EGBERTS, E. and TRAUB, P. Selective translation of mengovims RNA over host mRNA in homologous fractionated, cell-free translation systems from Ehrlich ascites tumor cells, Eur. J. of Biochem. (1978), 83, 555–561.
OBERG, B.R. and SHATKIN, A.J. Initiation of picornavirus protein synthesis in ascites cell extracts. Proc. Nat. Acad. Sci. U.S.A. (1972), 69, 3589–5595.
SMITH, A.E. The initiation of protein synthesis directed by the RNA from encephalomyocarditis virus. Eur. J. Biochem. (1973), 33, 301–313.
CELMA, M.L. and EHRENPELD, E. Translation of poliovirus RNA in vitro: Detection of two different initiation sites. J. Mol. Biol. (1975), 98, 761–780.
TERSHACZ, D.R, Effect of hypertonic medium on protein synthesis of Mahoney and LSc poliovirus. J. Virol. (1976), 20, 597–603.
NUSS, D.L., OPPEEMAN, H. and KOCH, G. Selective blockage of initiation of host protein synthesis in RNA virus-infected cells. Proc. Nat. Acad. Sci. U.S.A. (1975), 72, 1258–1262.
JENSE, H., KNAUERT, P. and EHRENFELD, E. Two initiation sites for translation of poliovirus RNA in vitro: Comparison of LSc and Mahoney
COLE, C.N., SMOLER, D., WIMMER, E. and BALTIMORE, D. Defective interfering particles of poliovirus. I. Isolation and physical properties. J. Virol. (1971), 7, 478–485.
COLE, C.N. and BALTIMORE, D. Defective interfering particles of poliovirus. II. Nature of the defect. J. Mol. Biol. (1973), 76, 323–343.
JACOBSON, M.F. and BALTIMORE, D. Morphogenesis of poliovirus. I. Association of viral RNA with coat protein. J. Mol. Biol. (1968), 33, 369–378.
BUTTERWORTH, B.E., HALL, L., STOLTZFUS, C.M. and RUECKERT, R.R. Virus-specific proteins synthesized in encephalomyocarditis virus-infected HeLa cells. Proc. Nat. Acad. Sci. U.S.A. (1971), 68, 3083–3087.
LUCAS-LENARD, J. Cleavage of mengovirus polyproteins in vivo. J. Virol. (1974), 14 261–269.
PAUCHA, E. and COLTER, J.S. Evidence for control of translation of the viral genome during replication of mengovirus and poliovirus. Virology (1975), 67, 300–305.
HELENTJARIS, T. and EHRENFELD, E. Inhibition of host cell protein synthesis by UV-inactivated poliovirus. J. Virol. (1977), 21, 259–267.
LUNDQUIST, R., EHRENFELD, E. and MAIZEL, J.V. Isolation of a viral polypeptide associated with poliovirus RNA polymerase. Proc. Nat. Acad. Sci. U.S.A. (1974), 71, 4773–4777.
LUNDQUIST, R. and MAIZEL, J.V. In vivo regulation of the poliovirus RNA polymerase. Virology (1978), 89, 484–493.
COLE, C.N. and BALTIMORE, D. Defective interfering particles of poliovirus. IV. Mechanisms of enrichment, J. Virol. (1973), 12, 1414–1426.
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Ehrenfeld, E. (1979). In Vitro Translation of Picornavirus RNA. In: Pérez-Bercoff, R. (eds) The Molecular Biology of Picornaviruses. NATO Advanced Study Institutes Series, vol 23. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-1000-6_11
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DOI: https://doi.org/10.1007/978-1-4684-1000-6_11
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