1887

Abstract

SUMMARY

Isolation of the Mengo virus stable non-capsid virus polypeptides E, F, G and I from infected L cells has been achieved. Unstable precursors were eliminated by incubation in the presence of pactamycin and capsid polypeptides were removed by ultracentrifugation and affinity chromatography. Subsequent sodium dodecyl sulphate (SDS)-hydroxylapatite chromatography resolved the non-capsid proteins into two major peaks which comprised F plus G and E plus I, respectively. The individual polypeptide species were separated by gel filtration on Sephadex G-100 in the presence of SDS.

Polypeptide E was isolated in an undenatured form by gel filtration of infected cell extracts (from which precursor and capsid polypeptides had been removed) on Bio-Gel A-5m agarose beads. Purified polypeptide E was found to co-sediment with Mengo virion RNA during centrifugation in a sucrose density gradient and it was also capable of binding to poly(A)-Sepharose. Assay mixtures containing polypeptide E exhibited an RNA polymerase activity which was dependent upon exogenous virus RNA template and oligo(U) primer and which was not affected by the addition of virus capsid polypeptides or extracts from uninfected cells.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-44-2-391
1979-08-01
2022-01-26
Loading full text...

Full text loading...

/deliver/fulltext/jgv/44/2/JV0440020391.html?itemId=/content/journal/jgv/10.1099/0022-1317-44-2-391&mimeType=html&fmt=ahah

References

  1. Baltimore D., Franklin R. M. 1963; A new ribonucleic acid polymerase appearing after Mengovirus infection of L-cells. Journal of Biological Chemistry 238:3395–3400
    [Google Scholar]
  2. Baltimore D., Eggers H. J., Franklin R. M., Tamm I. 1963; Poliovirus-induced RNA polymerase and the effects of virus-specific inhibitors on its production. Proceedings of the National Academy of Sciences of the United States of America 49:843–849
    [Google Scholar]
  3. Bowles S. A., Tershak D. R. 1978; Proteolysis of noncapsid protein 2 of type 3 poliovirus at the restrictive temperature: breakdown of non-capsid protein 2 correlates with loss of RNA synthesis. Journal of Virology 27:443–448
    [Google Scholar]
  4. Butterworth B. E., Hall L., Stoltzfus C. M., Rueckert R. R. 1971; Virus-specific proteins synthesized in encephalomyelitis virus-infected HeLa cells. Proceedings of the National Academy of Sciences of the United States of America 68:3083–3087
    [Google Scholar]
  5. Flanegan J. B., Baltimore D. 1977; Poliovirus-specific primer-dependent RNA polymerase able to copy poly(A). Proceedings of the National Academy of Sciences of the United States of America 74:3677–3680
    [Google Scholar]
  6. Flanegan J. B., Baltimore D. 1979; Poliovirus polyuridylic acid polymerase and RNA replicase have the same viral polypeptide. Journal of Virology 29:352–360
    [Google Scholar]
  7. Ghendon Y., Babushkina L., Blagoveshienskaya O. 1973; Inhibition of poliovirus RNA synthesis in an in vitro system by antiserum against 14S virus-specific structures. Archiv für die gesamte Virusforschung 40:47–51
    [Google Scholar]
  8. Gorbalenya A. E., Chumakov K. M., Agol V. I. 1978; RNA-binding properties of nonstructural polypeptide G of encephalomyocarditis virus. Virology 88:183–185
    [Google Scholar]
  9. Horton E., Liu S.-L., Martin E. M., Work T. S. 1966; Properties of a virus-induced RNA polymerase in ascites cells infected with encephalomyocarditis virus. Journal of Molecular Biology 15:62–76
    [Google Scholar]
  10. Korant B. D. 1972; Cleavage of viral precursor proteins in vivo and in vitro. Journal of Virology 10:751–759
    [Google Scholar]
  11. Korant B. D. 1973; Cleavage of poliovirus-specific polypeptide aggregates. Journal of Virology 12:556–563
    [Google Scholar]
  12. Korant B. D. 1975; Regulation of animal virus replication by protein cleavage. In Proteases and Biological Control pp 621–644 Edited by Reich E., Rifkin D. B., Shaw E. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  13. Korant B. D. 1977; Protein cleavage in virus-infected cells. Acta Biologica et Medica Germanica 36:1565–1573
    [Google Scholar]
  14. Loesch W. T., Arlinghaus R. B. 1975; Stable polypeptides associated the 250S Mengovirus-induced RNA polymerase structure. Archives of Virology 47:201–215
    [Google Scholar]
  15. Lundquist R. E., Ehrenfeld E., Maizel J. V. 1974; Isolation of a viral polypeptide associated with poliovirus RNA polymerase. Proceedings of the National Academy of Sciences of the United States of America 71:773–4777
    [Google Scholar]
  16. Mak T. W., O’Callaghan D. J., Kay C. M., Colter J. S. 1971; Studies of the protein subunit of pH-inactivated Mengo virus variants. II. Physico-chemical studies. Virology 43:579–587
    [Google Scholar]
  17. Paucha E., Seehafer J., Colter J. S. 1974; Synthesis of viral-specific polypeptides in Mengo virus-infected L cells: evidence for asymmetric translation of the viral genome. Virology 61:315–326
    [Google Scholar]
  18. Scraba D. G., Kay C. M., Colter J. S. 1967; Physico-chemical studies of three variants of Mengo virus and their constituent ribonucleates. Journal of Molecular Biology 26:67–79
    [Google Scholar]
  19. Summers D. G., Maizel J. V. 1968; Evidence for large precursor proteins in poliovirus synthesis. Proceedings of the National Academy of Sciences of the United States of America 59:966–971
    [Google Scholar]
  20. Taber R., Rekosh D., Baltimore D. 1971; Effect of pactamycin on synthesis of poliovirus proteins: a method for genetic mapping. Journal of Virology 8:395–401
    [Google Scholar]
  21. Traub A., Diskin B., Rosenbergh H., Kalmar E. 1976; Isolation and properties of the replicase of encephalomyocarditis virus. Journal of Virology 18:375–382
    [Google Scholar]
  22. Yin F. H. 1977; Involvement of viral procapsid in the 1NA synthesis and maturation of poliovirus. Virology 82:299–307
    [Google Scholar]
  23. Ziola B. R., Scraba D. G. 1974; Structure of the Mengo virion. I. Polypeptide and ribonucleate components of the virus particle. Virology 57:531–542
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-44-2-391
Loading
/content/journal/jgv/10.1099/0022-1317-44-2-391
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error