Abstract
The interferon induced enzyme 2'5' oligoadenylate synthetase produces 2'5' pppA(pA)n the first discovered natural nucleotide with a 2'5' linkage. We describe a direct assay of this enzyme based on separation by thin layer chromatography (TLC) of the substrate ATP and the products 2'5' pppA(pA)n (n larger than or equal to 1). This technique presents obvious advantages compared to the currently used methods. Moreover the enzyme uses other nucleotides as substrates forming co-oligonucleotides 2'5 pppA(pA)n pN (N = U,G,C,dA,dG,dT and dC). Additional procedures are described using different developing solvent systems for the separation of the core-2'5' oligonucleotides (2'5' A(pA)npN) containing AMP-residues entirely and those with another nucleotide at the 2' end.
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- BORSOOK H., DEASY C. L., HAAGENSMIT A. J., KEIGHLEY G., LOWY P. H. Incorporation in vitro of labeled amino acids into proteins of rabbit reticulocytes. J Biol Chem. 1952 May;196(2):669–694. [PubMed] [Google Scholar]
- Baglioni C. Interferon-induced enzymatic activities and their role in the antriviral state. Cell. 1979 Jun;17(2):255–264. doi: 10.1016/0092-8674(79)90151-x. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Gupta R. C., Randerath K. Use of specific endonuclease cleavage in RNA sequencing. Nucleic Acids Res. 1977 Jun;4(6):1957–1978. doi: 10.1093/nar/4.6.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Kerr I. M., Brown R. E., Hovanessian A. G. Nature of inhibitor of cell-free protein synthesis formed in response to interferon and double-stranded RNA. Nature. 1977 Aug 11;268(5620):540–542. doi: 10.1038/268540a0. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Randerath E., Randerath K. Ion-exchange thin-layer chromatography. XVI. Techniques for preparation and analysis of oligonucleotides. J Chromatogr. 1967 Dec;31(2):485–499. doi: 10.1016/s0021-9673(01)86099-4. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]