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. 1979 Oct;76(10):4778–4782. doi: 10.1073/pnas.76.10.4778

Interferon, double-stranded RNA, and RNA degradation: activation of an endonuclease by (2'-5')An.

E Slattery, N Ghosh, H Samanta, P Lengyel
PMCID: PMC413020  PMID: 291897

Abstract

Among the mediators of interferon action are one enzyme that is activated by double-stranded RNA to convert ATP to (2'-5')An and a second enzyme, an endonuclease, that is activated by (2'-5')An to cleave single-stranded RNA. The binding of (2'-5')An to the endonuclease (partially purified from mouse Ehrlich ascites tumor cells) is revealed by its retention on nitrocellulose filters. This can serve as the basis for an assay of the enzyme. Activation of the enzyme is reversible and is lost upon removal of (2'-5')An:gel filtration of activated endonuclease on Sephacryl S-200 results in an inactive enzyme. The enzyme can be activated again, however, by addition of (2'-5')An. The elution volume of the nonactivated endonuclease from Sephadex G-200 indicates that its molecular weight is 185,000, unusually large for a nuclease. The elution volume of the maximally activated endonuclease from Sephadex G-200 equilibrated with (2'-5')An is not detectably different from that of enzyme that had not been previously activated that was passed through Sephadex G-200 not equilibrated with (2'-5')An. This indicates that the activation does not result in a large change in the size or conformation of the enzyme.

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Selected References

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

  1. Baglioni C., Maroney P. A., West D. K. 2'5'Oligo(A) polymerase activity and inhibition of viral RNA synthesis in interferon-treated HeLa cells. Biochemistry. 1979 May 1;18(9):1765–1770. doi: 10.1021/bi00576a020. [DOI] [PubMed] [Google Scholar]
  2. Baglioni C., Minks M. A., Maroney P. A. Interferon action may be mediated by activation of a nuclease by pppA2'p5'A2'p5'A. Nature. 1978 Jun 22;273(5664):684–687. doi: 10.1038/273684a0. [DOI] [PubMed] [Google Scholar]
  3. Ball L. A. Induction of 2'5'-oligoadenylate synthetase activity and a new protein by chick interferon. Virology. 1979 Apr 30;94(2):282–296. doi: 10.1016/0042-6822(79)90462-8. [DOI] [PubMed] [Google Scholar]
  4. Ball L. A., White C. N. Oligonucleotide inhibitor of protein synthesis made in extracts of interferon-treated chick embryo cells: comparison with the mouse low molecular weight inhibitor. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1167–1171. doi: 10.1073/pnas.75.3.1167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brown G. E., Lebleu B., Kawakita M., Shaila S., Sen G. C., Lengyel P. Increased endonuclease activity in an extract from mouse Ehrlich ascites tumor cells which had been treated with a partially purified interferon preparation: dependence of double-stranded RNA;. Biochem Biophys Res Commun. 1976 Mar 8;69(1):114–122. doi: 10.1016/s0006-291x(76)80280-x. [DOI] [PubMed] [Google Scholar]
  6. Clemens M. J., Williams B. R. Inhibition of cell-free protein synthesis by pppA2'p5'A2'p5'A: a novel oligonucleotide synthesized by interferon-treated L cell extracts. Cell. 1978 Mar;13(3):565–572. doi: 10.1016/0092-8674(78)90329-x. [DOI] [PubMed] [Google Scholar]
  7. Eppstein D. A., Samuel C. E. Mechanism of interferon action. Properties of an interferon-mediated ribonucleolytic activity from mouse L929 cells. Virology. 1978 Aug;89(1):240–251. doi: 10.1016/0042-6822(78)90056-9. [DOI] [PubMed] [Google Scholar]
  8. Farrell P. J., Sen G. C., Dubois M. F., Ratner L., Slattery E., Lengyel P. Interferon action: two distinct pathways for inhibition of protein synthesis by double-stranded RNA. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5893–5897. doi: 10.1073/pnas.75.12.5893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hovanessian A. G., Kerr I. M. Synthesis of an oligonucleotide inhibitor of protein synthesis in rabbit reticulocyte lysates analogous to that formed in extracts from interferon-treated cells. Eur J Biochem. 1978 Mar;84(1):149–159. doi: 10.1111/j.1432-1033.1978.tb12151.x. [DOI] [PubMed] [Google Scholar]
  10. Kerr I. M., Brown R. E. pppA2'p5'A2'p5'A: an inhibitor of protein synthesis synthesized with an enzyme fraction from interferon-treated cells. Proc Natl Acad Sci U S A. 1978 Jan;75(1):256–260. doi: 10.1073/pnas.75.1.256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Martin E. M., Birdsall N. J., Brown R. E., Kerr I. M. Enzymic synthesis, characterisation and nuclear-magnetic-resonance spectra of pppA2'p5'A2'p5'A and related oligonucleotides: comparison with chemically synthesised material. Eur J Biochem. 1979 Apr 2;95(2):295–307. doi: 10.1111/j.1432-1033.1979.tb12965.x. [DOI] [PubMed] [Google Scholar]
  12. Minks M. A., Benvin S., Maroney P. A., Baglioni C. Metabolic stability of 2' 5'oligo (A) and activity of 2' 5'oligo (A)-dependent endonuclease in extracts of interferon-treated and control HeLa cells. Nucleic Acids Res. 1979 Feb;6(2):767–780. doi: 10.1093/nar/6.2.767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Minks M. A., Benvin S., Maroney P. A., Baglioni C. Synthesis of 2'5'-oligo(A) in extracts of interferon-treated HeLa cells. J Biol Chem. 1979 Jun 25;254(12):5058–5064. [PubMed] [Google Scholar]
  14. Nilsen T. W., Baglioni C. Mechanism for discrimination between viral and host mRNA in interferon-treated cells. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2600–2604. doi: 10.1073/pnas.76.6.2600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ratner L., Sen G. C., Brown G. E., Lebleu B., Kawakita M., Cabrer B., Slattery E., Lengyel P. Interferon, double-stranded RNA and RNA degradation. Characteristics of an endonuclease activity. Eur J Biochem. 1977 Oct 3;79(2):565–577. doi: 10.1111/j.1432-1033.1977.tb11841.x. [DOI] [PubMed] [Google Scholar]
  16. Ratner L., Wiegand R. C., Farrell P. J., Sen G. C., Cabrer B., Lengyel P. Interferon, double-stranded RNA and RNA degradation. Fractionation of the endonucleaseINT system into two macromolecular components; role of a small molecule in nuclease activation. Biochem Biophys Res Commun. 1978 Apr 14;81(3):947–954. doi: 10.1016/0006-291x(78)91443-2. [DOI] [PubMed] [Google Scholar]
  17. Roberts W. K., Hovanessian A., Brown R. E., Clemens M. J., Kerr I. M. Interferon-mediated protein kinase and low-molecular-weight inhibitor of protein synthesis. Nature. 1976 Dec 2;264(5585):477–480. doi: 10.1038/264477a0. [DOI] [PubMed] [Google Scholar]
  18. Schmidt A., Zilberstein A., Shulman L., Federman P., Berissi H., Revel M. Interferon action: isolation of nuclease F, a translation inhibitor activated by interferon-induced (2'-5') oligo-isoadenylate. FEBS Lett. 1978 Nov 15;95(2):257–264. doi: 10.1016/0014-5793(78)81006-0. [DOI] [PubMed] [Google Scholar]
  19. Sen G. C., Desrosiers R., Ratner L., Shaila S., Brown G. E., Lebleu B., Slattery E., Kawakita M., Cabrer B., Taira H. Messenger RNA methylation, translation and degradation in extracts of interferon-treated cells. Tex Rep Biol Med. 1977;35:221–229. [PubMed] [Google Scholar]
  20. Sen G. C., Lebleu B., Brown G. E., Kawakita M., Slattery E., Lengyel P. Interferon, double-stranded RNA and mRNA degradation. Nature. 1976 Nov 25;264(5584):370–373. doi: 10.1038/264370a0. [DOI] [PubMed] [Google Scholar]
  21. Sen G. C., Shaila S., Lebleu B., Brown G. E., Desrosiers R. C., Lengyel P. Impairment of reovirus mRNA methylation in extracts of interferon-treated Ehrilich ascites tumor cells: further characteristics of the phenomenon. J Virol. 1977 Jan;21(1):69–83. doi: 10.1128/jvi.21.1.69-83.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Williams B. R., Kerr I. M., Gilbert C. S., White C. N., Ball L. A. Synthesis and breakdown of pppA2'p5'A2'p5'A and transient inhibiton of protein synthesis in extracts from interferon-treated and control cells. Eur J Biochem. 1978 Dec;92(2):455–462. doi: 10.1111/j.1432-1033.1978.tb12767.x. [DOI] [PubMed] [Google Scholar]
  23. Zilberstein A., Kimchi A., Schmidt A., Revel M. Isolation of two interferon-induced translational inhibitors: a protein kinase and an oligo-isoadenylate synthetase. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4734–4738. doi: 10.1073/pnas.75.10.4734. [DOI] [PMC free article] [PubMed] [Google Scholar]

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