1887

Abstract

SUMMARY

Poliovirus type 1, vaccine strain (LSc, 2ab), which is a temperature- and actinomycin D-sensitive mutant derived from type 1 Mahoney strain, was grown in HeLa cells in the presence of P and a low concentration of actinomycin D. Seven and a half h p.i., genome P-RNA was recovered from the purified virion. Analysis of RNase T1 digests of the RNA by two-dimensional gel electrophoresis revealed that three possible point mutation sites exist in the large and unique oligonucleotides in the fingerprint. Neither a capping structure nor a nucleotide such as pppNp, ppNp or pNp, was detected by ion exchange column chromatography at pH 5.0 after digestion of virion RNA with RNase T2. Instead, a P-labelled compound, which could be digested with Pronase or proteinase K, was eluted at the void volume of the column. Proteinase K digests of the P-labelled compound contained pUp or pU as a single labelled compound, when genome RNA was digested with RNase T2 or nuclease P1, respectively, before digestion with the proteinase.

Our data locate possible point mutation sites on the genome of a mutant strain (LSc, 2ab) of type 1 poliovirus and show that a protein (VPg) is covalently bound to the 5′-terminus of the RNA. The protein (VPg) of LSc, 2ab strain migrates faster than capsid protein VP4 (mol. wt. 7000 to 8000) in a polyacrylamide gel and is thus similar to the VPg of the wild-type virus.

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1979-10-01
2021-10-21
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References

  1. Ambros V., Baltimore D. 1978; Protein is linked to the 5′-end of poliovirus RNA by a phosphodiester linkage to tyrosine. Journal of Biological Chemistry 253:5263–5266
    [Google Scholar]
  2. Ambros V., Pettersson R. F., Baltimore D. 1978; An enzyme activity in uninfected cells that cleaves the linkage between poliovirion RNA and the 5′-terminal protein. Cell 15:1439–1466
    [Google Scholar]
  3. Burroughs J. N., Brown F. 1978; Presence of a covalently linked protein on calicivirus RNA. Journal of General Virology 41:443–446
    [Google Scholar]
  4. Cooper P. D. 1966; The inhibition of poliovirus growth by actinomycin D and the prevention of the inhibition by pretreatment of the cells with serum or insulin. Virology 28:663–678
    [Google Scholar]
  5. Detjen B. M., Lucas J., Wimmer E. 1978; Poliovirus single-stranded RNA and double-stranded RNA: differential infectivity in enucleate cells. Journal of Virology 27:582–586
    [Google Scholar]
  6. Dewachter R., Fiers W. 1972; Preparative two-dimensional polyacrylamide gel electrophoresis of 32P-labelled RNA. Analytical Biochemistry 49:184–197
    [Google Scholar]
  7. Fiszman M., Bucchini D., Oirard M. 1971; Purification of the Sabin strain of poliovirus type i through treatment with sarkozyl. Journal of Virology 7:687–689
    [Google Scholar]
  8. Fiszman M., Reynier M., Bucchini D., Girard M. 1972; Thermosensitive block of the Sabin strain of poliovirus type i. Journal of Virology 10:1143–1151
    [Google Scholar]
  9. Flanegan J. B., Pettersson R. F., Ambros V., Hewlett M. J., Baltimore D. 1977; Covalent linkage of a protein to a defined nucleotide sequence at the 5′-terminus of virion and replicative intermediate RNAs of poliovirus. Proceedings of the National Academy of Sciences of the United States of America 74:961–965
    [Google Scholar]
  10. Frisby D., Eaton M., Fellner P. 1976; Absence of 5′-terminal capping in encephalomyocarditis virus RNA. Nucleic Acids Research 3:2771–2787
    [Google Scholar]
  11. Furuichi Y., Miura K. 1975; A blocked structure at the 5′ terminus of mRNA from cytoplasmic poly-hedrosis virus. Nature, London 253:374–375
    [Google Scholar]
  12. Golini F., Nomoto A., Wimmer E. 1978; Genome-linked protein of picornaviruses. IV. Difference in VPgs of encephalomyocarditis virus and poliovirus as evidence that genome-linked proteins are virus-coded. Virology 89:112–118
    [Google Scholar]
  13. Harris T. J. R., Dunn J. J., Wimmer E. 1978; Identification of specific fragment containing the 5′ end of poliovirus RNA after ribonuclease III digestion. Nucleic Acids Research 5:4037–4054
    [Google Scholar]
  14. Hruby D. E., Roberts W. K. 1978; Encephalomyocarditis virus RNA. III. Presence of a genome associated protein. Journal of Virology 25:413–415
    [Google Scholar]
  15. Klootwijk J., Klein I., Zabel P., Van Kammen A. 1977; Cowpea mosaic virus RNAs have neither m7GpppN …… nor mono-, di- or triphosphates at their 5′ ends. Cell 11:73–82
    [Google Scholar]
  16. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  17. Lee Y. F., Wimmer E. 1976; ‘Fingerprinting’ high molecular weight RNA by two-dimensional gel electrophoresis: application to poliovirus RNA. Nucleic Acids Research 3:1647–1658
    [Google Scholar]
  18. Lee Y. F., Nomoto A., Wimmer E. 1976; The genome of poliovirus is an exceptional eukaryotic mRNA. Progress in Nucleic Acid Research and Molecular Biology 19:89–96
    [Google Scholar]
  19. Lee Y. F., Nomoto A., Detjen B. M., Wimmer E. 1977; A protein covalently linked to poliovirus genome RNA. Proceedings of the National Academy of Sciences of the United States of America 74:59–63
    [Google Scholar]
  20. Lee Y. F., Kitamura N., Nomoto A., Wimmer E. 1979; Sequence study of poliovirus RNA. IV. Nucleotide sequence complexities of poliovirus type 1, 2 and two type 1 defective interfering particle RNAs, and finger print of poliovirus type 3 genome. Journal of General Virology 44:311–322
    [Google Scholar]
  21. Lwoff A. 1969; Death and transfiguration of a problem. Bacteriological Reviews 33:390–403
    [Google Scholar]
  22. Nishimura S. 1972; Minor components in transfer RNA: their characterization, location, and function. Progress in Nucleic Acid Research and Molecular Biology 12:49–81
    [Google Scholar]
  23. Nomoto A., Lee Y. F., Wimmer E. 1976; The 5′ end of poliovirus mRNA is not capped with m7G(5′)ppp-(5′)Np. Proceedings of the National Academy of Sciences of the United States of America 73:375–380
    [Google Scholar]
  24. Nomoto A., Detjen B. M., Pozzatti R., Wimmer E. 1977a; The location of the polio genome protein in viral RNAs and its implications for RNA synthesis. Nature, London 268:208–213
    [Google Scholar]
  25. Nomoto A., Kitamura N., Golini F., Wimmer E. 1977b; Poliovirion RNA and polio mRNA are identical molecules with the exception of the 5′-terminal genome-linked protein VPg. Proceedings of the National Academy of Sciences of the United States of America 74:5345–5349
    [Google Scholar]
  26. Pettersson R. F., Flanegan J. B., Rose J. K., Baltimore D. 1977; 5′-terminal nucleotide sequences of polio virus polyribosomal RNA and virion RNA are identical. Nature, London 268:270–272
    [Google Scholar]
  27. Pettersson R. F., Ambros V., Baltimore D. 1978; Identification of a protein linked to nascent poliovirus RNA and to the poly (U) of negative strand RNA. Journal of Virology 27:357–365
    [Google Scholar]
  28. Rothberg P. G., Harris T. J. R., Nomoto A., Wimmer E. 1978; O4-(5′-uridylyl) tyrosine is the bond between the genome-linked protein and the RNA of poliovirus. Proceedings of the National Academy of Sciences of the United States of America 75:4868–4872
    [Google Scholar]
  29. Sangar D. V., Rowlands D. J., Harris T. J. R., Brown F. 1977; Protein covalently linked to foot-and-mouth disease virus RNA. Nature, London 268:648–650
    [Google Scholar]
  30. Schaffer F. L., Gordon M. 1966; Differential inhibitory effects of actinomycin D among strains of poliovirus. Journal of Bacteriology 91:2309–2316
    [Google Scholar]
  31. Shatkin A. 1976; Capping of eucaryotic mRNAs. Cell 9:645–653
    [Google Scholar]
  32. Sulkowski E., Laskowski M. Sen 1971; Inactivation of 5′-nucleotidase in commercial preparations of venom exonuclease (phosphodiesterase). Biochimica et Biophysica Acta 240:443–447
    [Google Scholar]
  33. Wimmer E., Chang A. Y., Clark J. M., Reichmann M. E. 1968; Sequence studies of satellite tobacco necrosis virus RNA. Isolation and characterization of a 5′-terminal trinucleotide. Journal of Molecular Biology 38:59–73
    [Google Scholar]
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