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Radioimmune, radiobinding and HPLC analysis of 2-5A and related oligonucleotides from intact cells

Nature volume 288, pages 189–192 (1980)Cite this article

Abstract

The enzyme (2-5 A synthetase) which synthesizes ppp(A2′p)n A where n = 2 to 4 (collectively referred to as 2-5A)1 is widely distributed in a variety of cells and tissues in amounts which increase in response to interferon2–5 and vary with growth and hormone status6. 2-5A activates a nuclease which inhibits protein synthesis7–10. The non-phosphorylated ‘core’ of 2-5A ((A2′p)nA, n = 2 to 4) can inhibit DNA synthesis and cell growth11,12. Here we describe convenient and sensitive radioimmune (RI) and radiobinding (RB) assays for core and 2-5A. In combination with more satisfactory high performance liquid chromatography (HPLC) methods using reverse-phase C18 columns, these assays have been used to detect core and 2-5A in crude extracts from interferon-treated cells. The novel 2-5A synthetase products NAD2′p5′A2′p5′A and A5′p45′A2′p5′A2′p5′A (ref. 13), which can also be detected using the RB assay, were not found in significant amounts. The natural occurrence of core has not been described previously.

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Author notes

  1. M. Knight, P. J. Cayley, R. H. Silverman, C. S. Gilbert, R. E. Brown & I. M. Kerr

    Present address: ICRF Laboratories, Lincoln's Inn Fields, London, WC2A 3PX, UK

  2. D. H. Wreschner

    Present address: Department of Microbiology, Tel Aviv University, Ramat Aviv, Israel

Authors and Affiliations

  1. National Institute for Medical Research, Mill Hill, London, NW7 1AA, UK

    M. Knight, P. J. Cayley, R. H. Silverman, D. H. Wreschner, C. S. Gilbert, R. E. Brown & I. M. Kerr

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Knight, M., Cayley, P., Silverman, R. et al. Radioimmune, radiobinding and HPLC analysis of 2-5A and related oligonucleotides from intact cells. Nature 288, 189–192 (1980). https://doi.org/10.1038/288189a0

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