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. 1985 Sep;4(9):2249–2256. doi: 10.1002/j.1460-2075.1985.tb03922.x

Structure of two forms of the interferon-induced (2'-5') oligo A synthetase of human cells based on cDNAs and gene sequences.

P Benech, Y Mory, M Revel, J Chebath
PMCID: PMC554493  PMID: 2416561

Abstract

The (2'-5') oligo A synthetase E, one of the translational inhibitory enzymes whose synthesis is strongly induced by all interferons (IFNs), is shown to be encoded in human cells by a 13.5-kb gene. By a cell-specific differential splicing, between the seventh and an additional eighth exon of this gene, two active E mRNAs of 1.6 and 1.8 kb are produced, along with several longer transcripts. cDNA clones for the two mRNAs were obtained and their sequences indicate that the human (2'-5') oligo A synthetase gene codes for two forms of the enzyme of mol. wt. 41 000 and 46 000, which differ only by their C-terminal ends. The product of the 1.6-kb RNA (E16) has a very hydrophobic C terminus, which is replaced by a longer acidic C-terminal sequence in the 1.8-kb RNA product (E18). The transcriptional start site of the gene was identified and 200 bp of the 5' flanking region were sequenced. A strong homology was found between this region of the IFN-activated (2'-5') oligo A synthetase gene and the corresponding region of the human fibroblast IFN-beta 1 gene, whose transcription is also stimulated by IFN priming. The gene has two polyadenylation sites which share a common undecanucleotide, but are used in a cell-specific manner to give rise to the 1.6- and 1.8-kb mRNAs.

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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., Nilsen T. W. Mechanisms of antiviral action of interferon. Interferon. 1983;5:23–42. [PubMed] [Google Scholar]
  2. Ball L. A. Induction, purification, and properties of 2'5' oligoadenylate synthetase. Ann N Y Acad Sci. 1980;350:486–496. doi: 10.1111/j.1749-6632.1980.tb20651.x. [DOI] [PubMed] [Google Scholar]
  3. Benech P., Merlin G., Revel M., Chebath J. 3' end structure of the human (2'-5') oligo A synthetase gene: prediction of two distinct proteins with cell type-specific expression. Nucleic Acids Res. 1985 Feb 25;13(4):1267–1281. doi: 10.1093/nar/13.4.1267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  5. Chebath J., Merlin G., Metz R., Benech P., Revel M. Interferon-induced 56,000 Mr protein and its mRNA in human cells: molecular cloning and partial sequence of the cDNA. Nucleic Acids Res. 1983 Mar 11;11(5):1213–1226. doi: 10.1093/nar/11.5.1213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Creasey A. A., Eppstein D. A., Marsh Y. V., Khan Z., Merigan T. C. Growth regulation of melanoma cells by interferon and (2'-5')oligoadenylate synthetase. Mol Cell Biol. 1983 May;3(5):780–786. doi: 10.1128/mcb.3.5.780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Degrave W., Derynck R., Tavernier J., Haegeman G., Fiers W. Nucleotide sequence of the chromosomal gene for human fibroblast (beta 1) interferon and of the flanking regions. Gene. 1981 Aug;14(3):137–143. doi: 10.1016/0378-1119(81)90109-8. [DOI] [PubMed] [Google Scholar]
  8. Dougherty J. P., Samanta H., Farrell P. J., Lengyel P. Interferon, double-stranded RNA, and RNA degradation. Isolation of homogeneous pppA(2'p5'A)n-1 synthetase from Ehrlich ascites tumor cells. J Biol Chem. 1980 May 10;255(9):3813–3816. [PubMed] [Google Scholar]
  9. Epstein D. A., Czarniecki C. W., Jacobsen H., Friedman R. M., Panet A. A mouse cell line, which is unprotected by interferon against lytic virus infection, lacks ribonuclease F activity. Eur J Biochem. 1981 Aug;118(1):9–15. doi: 10.1111/j.1432-1033.1981.tb05479.x. [DOI] [PubMed] [Google Scholar]
  10. Fellous M., Nir U., Wallach D., Merlin G., Rubinstein M., Revel M. Interferon-dependent induction of mRNA for the major histocompatibility antigens in human fibroblasts and lymphoblastoid cells. Proc Natl Acad Sci U S A. 1982 May;79(10):3082–3086. doi: 10.1073/pnas.79.10.3082. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ferbus D., Justesen J., Besançon F., Thang M. N. The 2'5' oligoadenylate synthetase has a multifunctional 2'5' nucleotidyl-transferase activity. Biochem Biophys Res Commun. 1981 May 29;100(2):847–856. doi: 10.1016/s0006-291x(81)80251-3. [DOI] [PubMed] [Google Scholar]
  12. Friedman R. L., Manly S. P., McMahon M., Kerr I. M., Stark G. R. Transcriptional and posttranscriptional regulation of interferon-induced gene expression in human cells. Cell. 1984 Oct;38(3):745–755. doi: 10.1016/0092-8674(84)90270-8. [DOI] [PubMed] [Google Scholar]
  13. Friedman R. L., Stark G. R. alpha-Interferon-induced transcription of HLA and metallothionein genes containing homologous upstream sequences. Nature. 1985 Apr 18;314(6012):637–639. doi: 10.1038/314637a0. [DOI] [PubMed] [Google Scholar]
  14. Green M. R., Maniatis T., Melton D. A. Human beta-globin pre-mRNA synthesized in vitro is accurately spliced in Xenopus oocyte nuclei. Cell. 1983 Mar;32(3):681–694. doi: 10.1016/0092-8674(83)90054-5. [DOI] [PubMed] [Google Scholar]
  15. Johnston M. I., Friedman R. M., Torrence P. F. Interferon-induced (2'-5')oligoadenylate synthetase: adsorpton to and assay on adenosine 2',5'-diphosphate-sepharose. Biochemistry. 1980 Nov 25;19(24):5580–5585. doi: 10.1021/bi00565a019. [DOI] [PubMed] [Google Scholar]
  16. Karin M., Richards R. I. Human metallothionein genes--primary structure of the metallothionein-II gene and a related processed gene. Nature. 1982 Oct 28;299(5886):797–802. doi: 10.1038/299797a0. [DOI] [PubMed] [Google Scholar]
  17. Keller W. The RNA lariat: a new ring to the splicing of mRNA precursors. Cell. 1984 Dec;39(3 Pt 2):423–425. doi: 10.1016/0092-8674(84)90449-5. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. Kimchi A., Shure H., Lapidot Y., Rapoport S., Panet A., Revel M. Antimitogenic effects of interferon and (2'-5')-oligoadenylate in synchronized 3T3 fibroblasts. FEBS Lett. 1981 Nov 16;134(2):212–216. doi: 10.1016/0014-5793(81)80604-7. [DOI] [PubMed] [Google Scholar]
  21. Kitamura N., Semler B. L., Rothberg P. G., Larsen G. R., Adler C. J., Dorner A. J., Emini E. A., Hanecak R., Lee J. J., van der Werf S. Primary structure, gene organization and polypeptide expression of poliovirus RNA. Nature. 1981 Jun 18;291(5816):547–553. doi: 10.1038/291547a0. [DOI] [PubMed] [Google Scholar]
  22. Kozak M. Point mutations close to the AUG initiator codon affect the efficiency of translation of rat preproinsulin in vivo. Nature. 1984 Mar 15;308(5956):241–246. doi: 10.1038/308241a0. [DOI] [PubMed] [Google Scholar]
  23. Krishnan I., Baglioni C. Increased levels of (2'-5')oligo(A) polymerase activity in human lymphoblastoid cells treated with glucocorticoids. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6506–6510. doi: 10.1073/pnas.77.11.6506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  25. Laurence L., Marti J., Roux D., Cailla H. Immunological evidence for the in vivo occurrence of (2'-5')adenylyladenosine oligonucleotides in eukaryotes and prokaryotes. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2322–2326. doi: 10.1073/pnas.81.8.2322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lengyel P. Biochemistry of interferons and their actions. Annu Rev Biochem. 1982;51:251–282. doi: 10.1146/annurev.bi.51.070182.001343. [DOI] [PubMed] [Google Scholar]
  27. Malissen M., Malissen B., Jordan B. R. Exon/intron organization and complete nucleotide sequence of an HLA gene. Proc Natl Acad Sci U S A. 1982 Feb;79(3):893–897. doi: 10.1073/pnas.79.3.893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Maniatis T., Hardison R. C., Lacy E., Lauer J., O'Connell C., Quon D., Sim G. K., Efstratiadis A. The isolation of structural genes from libraries of eucaryotic DNA. Cell. 1978 Oct;15(2):687–701. doi: 10.1016/0092-8674(78)90036-3. [DOI] [PubMed] [Google Scholar]
  29. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  30. Merlin G., Chebath J., Benech P., Metz R., Revel M. Molecular cloning and sequence of partial cDNA for interferon-induced (2'-5')oligo(A) synthetase mRNA from human cells. Proc Natl Acad Sci U S A. 1983 Aug;80(16):4904–4908. doi: 10.1073/pnas.80.16.4904. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Minks M. A., West D. K., Benvin S., Baglioni C. Structural requirements of double-stranded RNA for the activation of 2',5'-oligo(A) polymerase and protein kinase of interferon-treated HeLa cells. J Biol Chem. 1979 Oct 25;254(20):10180–10183. [PubMed] [Google Scholar]
  32. Mory Y., Chernajovsky Y., Feinstein S. I., Chen L., Nir U., Weissenbach J., Malpiece Y., Tiollais P., Marks D., Ladner M. Synthesis of human interferon beta 1 in Escherichia coli infected by a lambda phage recombinant containing a human genomic fragment. Eur J Biochem. 1981 Nov;120(1):197–202. doi: 10.1111/j.1432-1033.1981.tb05689.x. [DOI] [PubMed] [Google Scholar]
  33. Nilsen T. W., Maroney P. A., Robertson H. D., Baglioni C. Heterogeneous nuclear RNA promotes synthesis of (2',5')oligoadenylate and is cleaved by the (2',5')oligoadenylate-activated endoribonuclease. Mol Cell Biol. 1982 Feb;2(2):154–160. doi: 10.1128/mcb.2.2.154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Nilsen T. W., Wood D. L., Baglioni C. Presence of 2',5'-oligo(A) and of enzymes that synthesize, bind, and degrade 2',5'-oligo(A) in HeLa cell nuclei. J Biol Chem. 1982 Feb 25;257(4):1602–1605. [PubMed] [Google Scholar]
  35. Nir U., Cohen B., Chen L., Revel M. A human IFN-beta 1 gene deleted of promoter sequences upstream from the TATA box is controlled post-transcriptionally by dsRNA. Nucleic Acids Res. 1984 Sep 25;12(18):6979–6993. doi: 10.1093/nar/12.18.6979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pustell J., Kafatos F. C. A convenient and adaptable package of DNA sequence analysis programs for microcomputers. Nucleic Acids Res. 1982 Jan 11;10(1):51–59. doi: 10.1093/nar/10.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Pustell J., Kafatos F. C. A high speed, high capacity homology matrix: zooming through SV40 and polyoma. Nucleic Acids Res. 1982 Aug 11;10(15):4765–4782. doi: 10.1093/nar/10.15.4765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Raj N. B., Pitha P. M. Analysis of interferon mRNA in human fibroblast cells induced to produce interferon. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7426–7430. doi: 10.1073/pnas.78.12.7426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Revel M., Kimchi A., Shulman L., Fradin A., Shuster R., Yakobson E., Chernajovsky Y., Schmidt A., Shure A., Bendori R. Role of interferon-induced enzymes in the antiviral and antimitogenic effects of interferon. Ann N Y Acad Sci. 1980;350:459–472. doi: 10.1111/j.1749-6632.1980.tb20649.x. [DOI] [PubMed] [Google Scholar]
  40. Revel M., Wallach D., Merlin G., Schattner A., Schmidt A., Wolf D., Shulman L., Kimchi A. Interferon-induced enzymes: microassays and their applications: purification and assay of (2'-5')-oligoadenylate synthetase and assay of 2'-phosphodiesterase. Methods Enzymol. 1981;79(Pt B):149–161. doi: 10.1016/s0076-6879(81)79024-4. [DOI] [PubMed] [Google Scholar]
  41. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  42. Rosa F., Berissi H., Weissenbach J., Maroteaux L., Fellous M., Revel M. The beta2-microglobulin mRNA in human Daudi cells has a mutated initiation codon but is still inducible by interferon. EMBO J. 1983;2(2):239–243. doi: 10.1002/j.1460-2075.1983.tb01412.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Rosa F., Hatat D., Abadie A., Wallach D., Revel M., Fellous M. Differential regulation of HLA-DR mRNAs and cell surface antigens by interferon. EMBO J. 1983;2(9):1585–1589. doi: 10.1002/j.1460-2075.1983.tb01628.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Salzberg S., Wreschner D. H., Oberman F., Panet A., Bakhanashvili M. Isolation and characterization of an interferon-resistant cell line deficient in the induction of (2'-5')oligoadenylate synthetase activity. Mol Cell Biol. 1983 Oct;3(10):1759–1765. doi: 10.1128/mcb.3.10.1759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Samanta H., Dougherty J. P., Lengyel P. Synthesis of (2'-5')(A)n from ATP. Characteristics of the reaction catalyzed by (2'-5')(A)n synthetase purified from mouse Ehrlich ascites tumor cells treated with interferon. J Biol Chem. 1980 Oct 25;255(20):9807–9813. [PubMed] [Google Scholar]
  46. Samanta H., Pravtcheva D. D., Ruddle F. H., Lengyel P. Chromosomal location of mouse gene 202 which is induced by interferons and specifies a 56.5 kD protein. J Interferon Res. 1984 Spring;4(2):295–300. doi: 10.1089/jir.1984.4.295. [DOI] [PubMed] [Google Scholar]
  47. Schamboeck A., Korman A. J., Kamb A., Strominger J. L. Organization of the transcriptional unit of a human class II histocompatibility antigen: HLA-DR heavy chain. Nucleic Acids Res. 1983 Dec 20;11(24):8663–8675. doi: 10.1093/nar/11.24.8663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. 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]
  49. Smekens-Etienne M., Goldstein J., Ooms H. A., Dumont J. E. Variation of (2'-5')oligo(adenylate) synthetase activity during rat-liver regeneration. Eur J Biochem. 1983 Feb 1;130(2):269–273. doi: 10.1111/j.1432-1033.1983.tb07146.x. [DOI] [PubMed] [Google Scholar]
  50. St Laurent G., Yoshie O., Floyd-Smith G., Samanta H., Sehgal P. B., Lengyel P. Interferon action: two (2'-5')(A)n synthetases specified by distinct mRNAs in Ehrlich ascites tumor cells treated with interferon. Cell. 1983 May;33(1):95–102. doi: 10.1016/0092-8674(83)90338-0. [DOI] [PubMed] [Google Scholar]
  51. 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]
  52. Strachan T., Sodoyer R., Damotte M., Jordan B. R. Complete nucleotide sequence of a functional class I HLA gene, HLA-A3: implications for the evolution of HLA genes. EMBO J. 1984 Apr;3(4):887–894. doi: 10.1002/j.1460-2075.1984.tb01901.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wallach D., Revel M. An interferon-induced cellular enzyme is incorporated into virions. Nature. 1980 Sep 4;287(5777):68–70. doi: 10.1038/287068a0. [DOI] [PubMed] [Google Scholar]
  54. Watling D., Serafinowska H. T., Reese C. B., Kerr I. M. Analogue inhibitor of 2-5A action: effect on the interferon-mediated inhibition of encephalomyocarditis virus replication. EMBO J. 1985 Feb;4(2):431–436. doi: 10.1002/j.1460-2075.1985.tb03647.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Weil J., Epstein C. J., Epstein L. B., Sedmak J. J., Sabran J. L., Grossberg S. E. A unique set of polypeptides is induced by gamma interferon in addition to those induced in common with alpha and beta interferons. Nature. 1983 Feb 3;301(5899):437–439. doi: 10.1038/301437a0. [DOI] [PubMed] [Google Scholar]
  56. Wigler M., Sweet R., Sim G. K., Wold B., Pellicer A., Lacy E., Maniatis T., Silverstein S., Axel R. Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell. 1979 Apr;16(4):777–785. doi: 10.1016/0092-8674(79)90093-x. [DOI] [PubMed] [Google Scholar]
  57. Wolf D., Rotter V. Major deletions in the gene encoding the p53 tumor antigen cause lack of p53 expression in HL-60 cells. Proc Natl Acad Sci U S A. 1985 Feb;82(3):790–794. doi: 10.1073/pnas.82.3.790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Yang K., Samanta H., Dougherty J., Jayaram B., Broeze R., Lengyel P. Interferons, double-stranded RNA, and RNA degradation. Isolation and characterization of homogeneous human (2'-5')(a)n synthetase. J Biol Chem. 1981 Sep 10;256(17):9324–9328. [PubMed] [Google Scholar]
  59. Yarden A., Shure-Gottlieb H., Chebath J., Revel M., Kimchi A. Autogenous production of interferon-beta switches on HLA genes during differentiation of histiocytic lymphoma U937 cells. EMBO J. 1984 May;3(5):969–973. doi: 10.1002/j.1460-2075.1984.tb01915.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. 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]
  61. Zinn K., DiMaio D., Maniatis T. Identification of two distinct regulatory regions adjacent to the human beta-interferon gene. Cell. 1983 Oct;34(3):865–879. doi: 10.1016/0092-8674(83)90544-5. [DOI] [PubMed] [Google Scholar]

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