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. 1981 Apr 10;9(7):1571–1581. doi: 10.1093/nar/9.7.1571

Ribosomal RNA cleavage, nuclease activation and 2-5A(ppp(A2'p)nA) in interferon-treated cells.

D H Wreschner, T C James, R H Silverman, I M Kerr
PMCID: PMC326782  PMID: 6164990

Abstract

Ribosomal RNA (rRNA) in intact ribosomes is cleaved into discrete products on incubation of reticulocyte lysates or L-cell extracts with ppp(A2'p)3A. Cleavage of rRNA may, therefore, provide a useful assay for 2-5A (ppp)A2'p)nA; n = 2 to 4) or for the presence of a 2-5A-dependent nuclease. The results with reticulocyte lysates differed from those obtained in the L-cell-free system in that (a) a different RNA cleavage pattern was produced (with added L-cell ribosomes) and (b) cleavage was fully activated by the analogue ppp(A2'p)3A3'pCp. As might be expected from the relatively high levels of 2-5A present in interferon-treated, encephalomyocarditis virus (EMC)-infected L-cells, rRNA extracted from these cells was also cleaved. The cleavage pattern observed overlapped with that obtained on incubation of an L-cell-free system with 2-5A. Thus, not only is 2-5A present, but the 2-5A-dependent nuclease also appears to be active, in interferon-treated, EMC-infected L-cells.

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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., 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]
  2. Bowman C. M., Sidikaro J., Nomura M. Specific inactivation of ribosomes by colicin E3 in vitro and mechanism of immunity in colicinogenic cells. Nat New Biol. 1971 Dec 1;234(48):133–137. doi: 10.1038/newbio234133a0. [DOI] [PubMed] [Google Scholar]
  3. Clemens M. J., Henshaw E. C., Rahamimoff H., London I. M. Met-tRNAfMet binding to 40S ribosomal subunits: a site for the regulation of initiation of protein synthesis by hemin. Proc Natl Acad Sci U S A. 1974 Aug;71(8):2946–2950. doi: 10.1073/pnas.71.8.2946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Hovanessian A. G., Wood J., Meurs E., Montagnier L. Increased nuclease activity in cells treated with pppA2'p5'A2'p5' A. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3261–3265. doi: 10.1073/pnas.76.7.3261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hunt T., Vanderhoff G., London I. M. Control of globin synthesis: the role of heme. J Mol Biol. 1972 May 28;66(3):471–481. doi: 10.1016/0022-2836(72)90427-5. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Knight M., Cayley P. J., Silverman R. H., Wreschner D. H., Gilbert C. S., Brown R. E., Kerr I. M. Radioimmune, radiobinding and HPLC analysis of 2-5A and related oligonucleotides from intact cells. Nature. 1980 Nov 13;288(5787):189–192. doi: 10.1038/288189a0. [DOI] [PubMed] [Google Scholar]
  9. Laskey R. A. The use of intensifying screens or organic scintillators for visualizing radioactive molecules resolved by gel electrophoresis. Methods Enzymol. 1980;65(1):363–371. doi: 10.1016/s0076-6879(80)65047-2. [DOI] [PubMed] [Google Scholar]
  10. Levy H. B., Riley F. L. Effect of interferon treatment on cellular messenger RNA. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3815–3819. doi: 10.1073/pnas.70.12.3815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Maroun L. E. Interferon effect on ribosomal ribonucleic acid related to chromosome 21 ploidy. Biochem J. 1979 Apr 1;179(1):221–225. doi: 10.1042/bj1790221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Maroun L. E. Interferon-mediated effect on ribosomal RNA metabolism. Biochim Biophys Acta. 1978 Jan 26;517(1):109–114. doi: 10.1016/0005-2787(78)90038-2. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Slattery E., Ghosh N., Samanta H., Lengyel P. Interferon, double-stranded RNA, and RNA degradation: activation of an endonuclease by (2'-5')An. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4778–4782. doi: 10.1073/pnas.76.10.4778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Stark G. R., Dower W. J., Schimke R. T., Brown R. E., Kerr I. M. 2-5A synthetase: assay, distribution and variation with growth or hormone status. Nature. 1979 Mar 29;278(5703):471–473. doi: 10.1038/278471a0. [DOI] [PubMed] [Google Scholar]
  18. Williams B. R., Golgher R. R., Brown R. E., Gilbert C. S., Kerr I. M. Natural occurrence of 2-5A in interferon-treated EMC virus-infected L cells. Nature. 1979 Dec 6;282(5739):582–586. doi: 10.1038/282582a0. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Wreschner D. H., McCauley J. W., Skehel J. J., Kerr I. M. Interferon action--sequence specificity of the ppp(A2'p)nA-dependent ribonuclease. Nature. 1981 Jan 29;289(5796):414–417. doi: 10.1038/289414a0. [DOI] [PubMed] [Google Scholar]
  21. Wreschner D., Melloul D., Herzberg M. Interaction between membrane functions and protein synthesis in reticulocytes. Isolation of RNase M, a membrane component inhibiting protein synthesis through specific endonucleolytic activity. Eur J Biochem. 1978 Sep 1;89(2):341–352. doi: 10.1111/j.1432-1033.1978.tb12535.x. [DOI] [PubMed] [Google Scholar]

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