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Van Dyke TA, Flanegan JB. Identification of poliovirus polypeptide P63 as a soluble RNA-dependent RNA polymerase. J Virol. 1980;35(3):732-40doi: 10.1128/JVI.35.3.732-740.1980.
Van Dyke, T. A., & Flanegan, J. B. (1980). Identification of poliovirus polypeptide P63 as a soluble RNA-dependent RNA polymerase. Journal of virology, 35(3), 732-40. https://doi.org/10.1128/JVI.35.3.732-740.1980
Van Dyke, T A, and Flanegan, J B. "Identification of poliovirus polypeptide P63 as a soluble RNA-dependent RNA polymerase." Journal of virology vol. 35,3 (1980): 732-40. doi: https://doi.org/10.1128/JVI.35.3.732-740.1980
Van Dyke TA, Flanegan JB. Identification of poliovirus polypeptide P63 as a soluble RNA-dependent RNA polymerase. J Virol. 1980 Sep;35(3):732-40. doi: 10.1128/JVI.35.3.732-740.1980. PMID: 6252335; PMCID: PMC288867.
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Atypon Free PMC Article

J Virol. 1980 Sep;35(3):732-40. doi: 10.1128/JVI.35.3.732-740.1980.

Identification of poliovirus polypeptide P63 as a soluble RNA-dependent RNA polymerase.

Journal of virology

T A Van Dyke, J B Flanegan

PMID: 6252335 PMCID: PMC288867 DOI: 10.1128/JVI.35.3.732-740.1980
Free PMC Article

Abstract

A poliovirus-specific RNA-dependent RNA polymerase was isolated from a cytoplasmic extract of infected HeLa cells and was shown to copurify with a single virus-specific protein. The polymerase was isolated from cells labeled with [35S]-methionine and was fractionated from other soluble cytoplasmic proteins by ammonium sulfate precipitation, phosphocellulose chromatography, gel filtration on Sephacryl S-200, and chromatography on hydroxylapatite. The activity of the enzyme was measured by using either polyadenylic acid or poliovirion RNA as a template in the presence of an oligouridylic acid primer. A single virus-specific protein that had an apparent molecular weight of 63,000 (p63) was found to copurify with this activity. Host-coded proteins were present in reduced molar amounts relative to p63. Noncapsid viral protein 2 (NCVP2) and other viral proteins were clearly separated from p63 by gel filtration on Sephacryl S-200. Polymerase activity coeluted from the column precisely with p63. NCVP2 was totally inactive as an RNA polymerase and did not stimulate the polymerase activity of p63. The purified enzyme sedimented at about 4S on a glycerol gradient and thus appeared to be a monomer of p63. Two-dimensional gel electrophoresis of the polymerase protein indicated that it had an isoelectric point of about 7.5. Thus, the viral polypeptide, p63, as defined by the above physical parameters, is an RNA-dependent RNA polymerase that can copy poliovirion RNA when oligouridylic acid is used as a primer.

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References

  1. Nature. 1977 Jul 21;268(5617):208-13 - PubMed
  2. Virology. 1977 Jul 15;80(2):297-308 - PubMed
  3. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3677-80 - PubMed
  4. Virology. 1978 Apr;85(2):434-44 - PubMed
  5. Virology. 1978 Apr;85(2):445-55 - PubMed
  6. J Virol. 1978 Aug;27(2):443-8 - PubMed
  7. Virology. 1978 Sep;89(2):484-93 - PubMed
  8. J Virol. 1979 Jan;29(1):352-60 - PubMed
  9. J Biol Chem. 1979 Jun 25;254(12):5000-7 - PubMed
  10. Proc Natl Acad Sci U S A. 1966 Aug;56(2):741-8 - PubMed
  11. J Virol. 1969 Apr;3(4):376-84 - PubMed
  12. J Mol Biol. 1973 May 25;76(3):325-43 - PubMed
  13. Biochem J. 1973 Jul;133(3):529-39 - PubMed
  14. Eur J Biochem. 1974 Jul 1;46(1):83-8 - PubMed
  15. Methods Enzymol. 1974;30:709-23 - PubMed
  16. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4773-7 - PubMed
  17. J Biol Chem. 1975 May 25;250(10):4007-21 - PubMed
  18. Proc Natl Acad Sci U S A. 1975 Oct;72(10):4157-61 - PubMed
  19. Anal Biochem. 1976 May 7;72:248-54 - PubMed
  20. J Virol. 1976 Aug;19(2):457-66 - PubMed
  21. Proc Natl Acad Sci U S A. 1977 Mar;74(3):961-5 - PubMed
  22. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2679-83 - PubMed
  23. Cell. 1980 Feb;19(2):423-9 - PubMed
  24. J Virol. 1979 Oct;32(1):155-61 - PubMed
  25. Proc Natl Acad Sci U S A. 1963 Jun;49:843-9 - PubMed

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