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. 1974 Jun;13(6):1338–1345. doi: 10.1128/jvi.13.6.1338-1345.1974

Poly(A) Polymerase Activity in Reovirus

C M Stoltzfus a,1, M Morgan a, A K Banerjee a, A J Shatkin a
PMCID: PMC355454  PMID: 4833612

Abstract

An enzymatic activity which synthesized oligo(A) in vitro was found in highly purified reovirus. The poly(A) polymerase activity was dependent on Mn2+ and utilized only ATP, whereas the virion-associated RNA polymerase required all four ribonucleoside triphosphates and Mg2+. Oligo(A) synthesis was demonstrated with complete virions and infectious subviral particles derived from virus by limited chymotrypsin digestion but not with cores, a product of extensive chymotrypsin digestion of virus. The enzymatic product and the oligo(A) from purified virions were isolated by binding to oligo(dT)-cellulose columns. Most of the in vitro product was similar in size and structure to the oligo(A) from purified virions by the criteria of gel electrophoresis, DEAE-cellulose chromatography, end-group analysis, and sensitivity to RNase. The evidence suggests that oligo(A) synthesis is mediated by the poly(A) polymerase during a late step in viral morphogenesis and may result from an alternative activity of the virion-associated transcriptase.

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Selected References

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

  1. Arima T., Uchida T., Egami F. Studies on extracellular ribonucleases of Ustilago sphaerogena. Characterization of substrate specificity with special reference to purine-specific ribonucleases. Biochem J. 1968 Feb;106(3):609–613. doi: 10.1042/bj1060609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BEERS R. F., Jr Hydrolysis of polyadenylic acid by pancreatic ribonuclease. J Biol Chem. 1960 Aug;235:2393–2398. [PubMed] [Google Scholar]
  4. Banerjee A. K., Rensing U., August J. T. Replication of RNA viruses. X. Replication of a natural 6 s RNA by the Q-beta RNA polymerase. J Mol Biol. 1969 Oct 28;45(2):181–193. doi: 10.1016/0022-2836(69)90098-9. [DOI] [PubMed] [Google Scholar]
  5. Banerjee A. K., Rhodes D. P. In vitro synthesis of RNA that contains polyadenylate by virion-associated RNA polymerase of vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3566–3570. doi: 10.1073/pnas.70.12.3566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Banerjee A. K., Shatkin A. J. Guanosine-5'-diphosphate at the 5' termini of reovirus RNA: evidence for a segmented genome within the virion. J Mol Biol. 1971 Nov 14;61(3):643–653. doi: 10.1016/0022-2836(71)90069-6. [DOI] [PubMed] [Google Scholar]
  7. Banerjee A. K., Shatkin A. J. Transcription in vitro by reovirus-associated ribonucleic acid-dependent polymerase. J Virol. 1970 Jul;6(1):1–11. doi: 10.1128/jvi.6.1.1-11.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Banerjee A. K., Ward R., Shatkin A. J. Initiation of reovirus mRNA synthesis in vitro. Nat New Biol. 1971 Apr 7;230(14):169–172. doi: 10.1038/newbio230169a0. [DOI] [PubMed] [Google Scholar]
  9. Bellamy A. R., Joklik W. K. Studies on the A-rich RNA of reovirus. Proc Natl Acad Sci U S A. 1967 Oct;58(4):1389–1395. doi: 10.1073/pnas.58.4.1389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Borsa J., Graham A. F. Reovirus: RNA polymerase activity in purified virions. Biochem Biophys Res Commun. 1968 Dec 30;33(6):895–901. doi: 10.1016/0006-291x(68)90396-3. [DOI] [PubMed] [Google Scholar]
  11. Borsa J., Grover J., Chapman J. D. Presence of nucleoside triphosphate phosphohydrolase activity in purified virions of reovirus. J Virol. 1970 Sep;6(3):295–302. doi: 10.1128/jvi.6.3.295-302.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Brown M., Dorson J. W., Bollum F. J. Terminal riboadenylate transferase: a poly A polymerase in purified vaccinia virus. J Virol. 1973 Aug;12(2):203–208. doi: 10.1128/jvi.12.2.203-208.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. CHAMBERLIN M., BERG P. MECHANISM OF RNA POLYMERASE ACTION: CHARACTERIZATION OF THE DNA-DEPENDENT SYNTHESIS OF POLYADENYLIC ACID. J Mol Biol. 1964 May;8:708–726. doi: 10.1016/s0022-2836(64)80120-0. [DOI] [PubMed] [Google Scholar]
  14. Carter C., Stoltzfus C. M., Banerjee A. K., Shatkin A. J. Origin of reovirus oligo(A). J Virol. 1974 Jun;13(6):1331–1337. doi: 10.1128/jvi.13.6.1331-1337.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Darnell J. E., Jelinek W. R., Molloy G. R. Biogenesis of mRNA: genetic regulation in mammalian cells. Science. 1973 Sep 28;181(4106):1215–1221. doi: 10.1126/science.181.4106.1215. [DOI] [PubMed] [Google Scholar]
  16. Graziadei W. D., 3rd, Roy D., Konigsberg W., Lengyel P. Translation of reovirus messenger ribonucleic acids synthesized in vitro into reovirus proteins in a mouse L cell extract. Arch Biochem Biophys. 1973 Sep;158(1):266–275. doi: 10.1016/0003-9861(73)90621-8. [DOI] [PubMed] [Google Scholar]
  17. Joklik W. K. Studies on the effect of chymotrypsin on reovirions. Virology. 1972 Sep;49(3):700–715. doi: 10.1016/0042-6822(72)90527-2. [DOI] [PubMed] [Google Scholar]
  18. Kapuler A. M. An extraordinary temperature dependence of the reovirus transcriptase. Biochemistry. 1970 Oct 27;9(22):4453–4457. doi: 10.1021/bi00824a029. [DOI] [PubMed] [Google Scholar]
  19. Kapuler A. M., Mendelsohn N., Klett H., Acs G. Four base-specific nucleoside 5'-triphosphatases in the subviral core of reovirus. Nature. 1970 Mar 28;225(5239):1209–1213. doi: 10.1038/2251209a0. [DOI] [PubMed] [Google Scholar]
  20. Koide F., Suzuka I., Sekiguchi K. Some properties of an adenine-rich polynucleotide fragment from the avian reovirus. Biochem Biophys Res Commun. 1968 Jan 11;30(1):95–99. doi: 10.1016/0006-291x(68)90718-3. [DOI] [PubMed] [Google Scholar]
  21. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  22. Levin D. H., Mendelsohn N., Schonberg M., Klett H., Silverstein S., Kapuler A. M., Acs G. Properties of RNA transcriptase in reovirus subviral particles. Proc Natl Acad Sci U S A. 1970 Jul;66(3):890–897. doi: 10.1073/pnas.66.3.890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McDowell M. J., Joklik W. K., Villa-Komaroff L., Lodish H. F. Translation of reovirus messenger RNAs synthetesized in vitro into reovirus polypeptides by several mammalian cell-free extracts. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2649–2653. doi: 10.1073/pnas.69.9.2649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Molloy G. R., Thomas W. L., Darnell J. E. Occurrence of uridylate-rich oligonucleotide regions in heterogeneous nuclear RNA of HeLa cells. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3684–3688. doi: 10.1073/pnas.69.12.3684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nichols J. L., Bellamy A. R., Joklik W. K. Identification of the nucleotide sequences of the oligonucleotides present in reovirions. Virology. 1972 Aug;49(2):562–572. doi: 10.1016/0042-6822(72)90507-7. [DOI] [PubMed] [Google Scholar]
  26. Perry R. P., La Torre J., Kelley D. E., Greenberg J. R. On the lability of poly(A) sequences during extraction of messenger RNA from polyribosomes. Biochim Biophys Acta. 1972 Mar 14;262(2):220–226. doi: 10.1016/0005-2787(72)90236-5. [DOI] [PubMed] [Google Scholar]
  27. Roeder R. G., Rutter W. J. Multiple forms of DNA-dependent RNA polymerase in eukaryotic organisms. Nature. 1969 Oct 18;224(5216):234–237. doi: 10.1038/224234a0. [DOI] [PubMed] [Google Scholar]
  28. STEVENS A. STUDIES OF THE RIBONUCLEIC ACID POLYMERASE FROM ESCHERICHIA COLI. II. STUDIES OF HOMOPOLYMER FORMATION. J Biol Chem. 1964 Jan;239:204–209. [PubMed] [Google Scholar]
  29. Schonberg M., Silverstein S. C., Levin D. H., Acs G. Asynchronous synthesis of the complementary strands of the reovirus genome. Proc Natl Acad Sci U S A. 1971 Feb;68(2):505–508. doi: 10.1073/pnas.68.2.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Shatkin A. J. Animal RNA viruses: genome structure and function. Annu Rev Biochem. 1974;43(0):643–665. doi: 10.1146/annurev.bi.43.070174.003235. [DOI] [PubMed] [Google Scholar]
  31. Shatkin A. J., LaFiandra A. J. Transcription by infectious subviral particles of reovirus. J Virol. 1972 Oct;10(4):698–706. doi: 10.1128/jvi.10.4.698-706.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shatkin A. J., Sipe J. D. RNA polymerase activity in purified reoviruses. Proc Natl Acad Sci U S A. 1968 Dec;61(4):1462–1469. doi: 10.1073/pnas.61.4.1462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Shatkin A. J., Sipe J. D. Single-stranded, adenine-rich RNA from purified reoviruses. Proc Natl Acad Sci U S A. 1968 Jan;59(1):246–253. doi: 10.1073/pnas.59.1.246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Silverstein S. C., Astell C., Christman J., Klett H., Acs G. Shythesis of reovirus oligo adenylic acid in vivo and in vitro. J Virol. 1974 Mar;13(3):740–752. doi: 10.1128/jvi.13.3.740-752.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Skehel J. J., Joklik W. K. Studies on the in vitro transcription of reovirus RNA catalyzed by reovirus cores. Virology. 1969 Dec;39(4):822–831. doi: 10.1016/0042-6822(69)90019-1. [DOI] [PubMed] [Google Scholar]
  36. Stoltzfus C. M., Banerjee A. K. Two oligonucleotide classes of single-stranded ribopolymers in reovirus A-rich RNA. Arch Biochem Biophys. 1972 Oct;152(2):733–743. doi: 10.1016/0003-9861(72)90269-x. [DOI] [PubMed] [Google Scholar]
  37. Villarreal L. P., Holland J. J. Synthesis of poly(A) in vitro by purified virions of vesicular stomatitis virus. Nat New Biol. 1973 Nov 7;246(149):17–19. doi: 10.1038/newbio246017a0. [DOI] [PubMed] [Google Scholar]
  38. Winters M. A., Edmonds M. A poly(A) polymerase from calf thymus. Purification and properities of the enzyme. J Biol Chem. 1973 Jul 10;248(13):4756–4762. [PubMed] [Google Scholar]

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