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. 1974 Dec;71(12):4773–4777. doi: 10.1073/pnas.71.12.4773

Isolation of a Viral Polypeptide Associated with Poliovirus RNA Polymerase

R E Lundquist 1,*, E Ehrenfeld 1, J V Maizel Jr 1,*,†
PMCID: PMC433979  PMID: 4373729

Abstract

Poliovirus-infected HeLa cells were labeled with radioactive methionine or phenylalanine and subjected to a new purification procedure for the viral induced RNA polymerase activity. Detergent-solubilized polymerase activity was purified by precipitation with 2 M LiCl and sedimentation through sucrose gradients. Approximately 0.001% of the incorporated amino acid radio-activity sediments with the peak of polymerase activity. Gradient fractions comprising the polymerase activity peak were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and found to contain predominantly one virus-specific polypeptide. Polyacrylamide gel electrophoresis also reveals that this purified polypeptide migrates with a 58,000 molecular weight noncapsid polio-virus polypeptide.

Keywords: picornavirus, replication complex, replicase

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arlinghaus R. B., Polatnick J. The isolation of two enzyme-ribonucleic acid complexes involved in the synthesis of foot-and-mouth disease virus ribonucleic acid. Proc Natl Acad Sci U S A. 1969 Mar;62(3):821–828. doi: 10.1073/pnas.62.3.821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arlinghaus R. B., Syrewicz J. J., Loesch W. T., Jr RNA polymerase complexes from mengovirus infected cells. Arch Gesamte Virusforsch. 1972;38(1):17–28. doi: 10.1007/BF01241352. [DOI] [PubMed] [Google Scholar]
  3. BALTIMORE D., EGGERS H. J., FRANKLIN R. M., TAMM I. Poliovirus-induced RNA polymerase and the effects of virus-specific inhibitors on its production. Proc Natl Acad Sci U S A. 1963 Jun;49:843–849. doi: 10.1073/pnas.49.6.843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. BALTIMORE D. IN VITRO SYNTHESIS OF VIRAL RNA BY THE POLIOVIRUS RNA POLYMERASE. Proc Natl Acad Sci U S A. 1964 Mar;51:450–456. doi: 10.1073/pnas.51.3.450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. BARLOW J. J., MATHIAS A. P., WILLIAMSON R., GAMMACK D. B. A SIMPLE METHOD FOR THE QUANTITATIVE ISOLATION OF UNDEGRADED HIGH MOLECULAR WEIGHT RIBONUCLEIC ACID. Biochem Biophys Res Commun. 1963 Sep 10;13:61–66. doi: 10.1016/0006-291x(63)90163-3. [DOI] [PubMed] [Google Scholar]
  6. Baltimore D., Girard M., Darnell J. E. Aspects of the synthesis of poliovirus RNA and the formation of virus particles. Virology. 1966 Jun;29(2):179–189. doi: 10.1016/0042-6822(66)90024-9. [DOI] [PubMed] [Google Scholar]
  7. Butterworth B. E. A comparison of the virus-specific polypeptides of encephalomyocarditis virus, human rhinovirus-1A, and poliovirus. Virology. 1973 Dec;56(2):439–453. doi: 10.1016/0042-6822(73)90048-2. [DOI] [PubMed] [Google Scholar]
  8. Caliguiri L. A., Mosser A. G. Proteins associated with the poliovirus RNA replication complex. Virology. 1971 Nov;46(2):375–386. doi: 10.1016/0042-6822(71)90039-0. [DOI] [PubMed] [Google Scholar]
  9. Caliguiri L. A., Tamm I. Action of guanidine on the replication of poliovirus RNA. Virology. 1968 Jul;35(3):408–417. doi: 10.1016/0042-6822(68)90219-5. [DOI] [PubMed] [Google Scholar]
  10. Caliguiri L. A., Tamm I. Membranous structures associated with translation and transcription of poliovirus RNA. Science. 1969 Nov 14;166(3907):885–886. doi: 10.1126/science.166.3907.885. [DOI] [PubMed] [Google Scholar]
  11. DALGARNO L., MARTIN E. M. STUDIES ON EMC VIRAL RNA SYNTHESIS AND ITS LOCALIZATION IN INFECTED KREBS ASCITES CELLS. Virology. 1965 Jul;26:450–465. doi: 10.1016/0042-6822(65)90008-5. [DOI] [PubMed] [Google Scholar]
  12. Ehrenfeld E., Maizel J. V., Summers D. F. Soluble RNA polymerase complex from poliovirus-infected HeLa cells. Virology. 1970 Apr;40(4):840–846. doi: 10.1016/0042-6822(70)90129-7. [DOI] [PubMed] [Google Scholar]
  13. Holland J. J., Kiehn E. D. Specific cleavage of viral proteins as steps in the synthesis and maturation of enteroviruses. Proc Natl Acad Sci U S A. 1968 Jul;60(3):1015–1022. doi: 10.1073/pnas.60.3.1015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jacobson M. F., Baltimore D. Polypeptide cleavages in the formation of poliovirus proteins. Proc Natl Acad Sci U S A. 1968 Sep;61(1):77–84. doi: 10.1073/pnas.61.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kamen R. Characterization of the subunits of Q-beta replicase. Nature. 1970 Nov 7;228(5271):527–533. doi: 10.1038/228527a0. [DOI] [PubMed] [Google Scholar]
  16. Kondo M., Gallerani R., Weissmann C. Subunit structure of Q-beta replicase. Nature. 1970 Nov 7;228(5271):525–527. doi: 10.1038/228525a0. [DOI] [PubMed] [Google Scholar]
  17. Levintow L., Bishop J. M. Comparative aspects of Poliovirus replication. J Cell Physiol. 1970 Dec;76(3):265–271. doi: 10.1002/jcp.1040760306. [DOI] [PubMed] [Google Scholar]
  18. Maizel J. V., Jr, Summers D. F. Evidence for differences in size and composition of the poliovirus-specific polypeptides in infected HeLa cells. Virology. 1968 Sep;36(1):48–54. doi: 10.1016/0042-6822(68)90115-3. [DOI] [PubMed] [Google Scholar]
  19. Penman S., Summers D. Effects on host cell metabolism following synchronous infection with poliovirus. Virology. 1965 Dec;27(4):614–620. doi: 10.1016/0042-6822(65)90187-x. [DOI] [PubMed] [Google Scholar]
  20. Plagemann P. G., Swim H. E. Symposium on replication of viral nucleic acids. 3. Replication of mengovirus ribonucleic acid. Bacteriol Rev. 1966 Jun;30(2):288–308. doi: 10.1128/br.30.2.288-308.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Staples D. H., Hindley J., Billeter M. A., Weissmann C. Localization of Q-beta maturation cistron ribosome binding site. Nat New Biol. 1971 Sep 15;234(50):202–204. doi: 10.1038/newbio234202a0. [DOI] [PubMed] [Google Scholar]
  22. Summers D. F., Maizel J. V., Jr Determination of the gene sequence of poliovirus with pactamycin. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2852–2856. doi: 10.1073/pnas.68.11.2852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Summers D. F., Maizel J. V., Jr Evidence for large precursor proteins in poliovirus synthesis. Proc Natl Acad Sci U S A. 1968 Mar;59(3):966–971. doi: 10.1073/pnas.59.3.966. [DOI] [PMC free article] [PubMed] [Google Scholar]

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