Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1996 Dec 1;320(Pt 2):519–530. doi: 10.1042/bj3200519

A novel trans-spliced mRNA from Onchocerca volvulus encodes a functional S-adenosylmethionine decarboxylase.

A A Da'Dara 1, K Henkle-Dührsen 1, R D Walter 1
PMCID: PMC1217960  PMID: 8973561

Abstract

Complete cDNA and genomic sequences encoding the Onchocerca volvulus S-adenosylmethionine decarboxylase (SAMDC), a key enzyme in polyamine biosynthesis, have been isolated and characterized. The deduced amino acid sequence encodes a 42 kDa proenzyme with a moderate level of sequence homology to eukaryotic SAMDCs. Enzymically active O. volvulus SAMDC was expressed at a high level in an Escherichia coli mutant strain lacking endogenous SAMDC. The recombinant enzyme was purified to homogeneity using DEAE-cellulose, methylglyoxal bis(guanylhydrazone)-Sepharose and Superdex S-200 chromatography. It was determined that the recombinant proenzyme is cleaved to produce 32 and 10 kDa subunits. The sequence of the N-terminal portion of the large subunit was determined and comparison with the sequence of the proenzyme revealed that the precise cleavage site lies between Glu86 and Ser87. Gel-filtration experiments demonstrated that these two subunits combine to form an active heterotetramer. Comparison of the cDNA and genomic sequences revealed that the SAMDC mRNA undergoes both cis- and trans-splicing in its 5'-untranslated region (UTR). Anchored PCR on O. volvulus mRNA confirmed the cDNA sequence and identified two distinct trans-spliced products, a 22-nucleotide spliced-leader sequence and a 138 bp sequence containing the 22 nucleotide spliced-leader sequence. Genomic Southern-blot analysis suggests that the O. volvulus SAMDC is encoded by a single-copy gene. This gene spans 5.3 kb and is comprised of nine exons and eight introns. The first intron is located in the 5'-UTR and processing of this intron has a potential regulatory function. The 5'-flanking region of the gene contains potential transcriptional regulatory elements such as a TATA box, two CAAT boxes and AP-1-, C/EBP-, ELP-, H-APF-1-, HNF-5- and PEA3-binding sites.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. Anton D. L., Kutny R. Escherichia coli S-adenosylmethionine decarboxylase. Subunit structure, reductive amination, and NH2-terminal sequences. J Biol Chem. 1987 Feb 25;262(6):2817–2822. [PubMed] [Google Scholar]
  2. Auvinen M., Paasinen A., Andersson L. C., Hölttä E. Ornithine decarboxylase activity is critical for cell transformation. Nature. 1992 Nov 26;360(6402):355–358. doi: 10.1038/360355a0. [DOI] [PubMed] [Google Scholar]
  3. Balasundaram D., Tabor C. W., Tabor H. Oxygen toxicity in a polyamine-depleted spe2 delta mutant of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4693–4697. doi: 10.1073/pnas.90.10.4693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bitonti A. J., Byers T. L., Bush T. L., Casara P. J., Bacchi C. J., Clarkson A. B., Jr, McCann P. P., Sjoerdsma A. Cure of Trypanosoma brucei brucei and Trypanosoma brucei rhodesiense infections in mice with an irreversible inhibitor of S-adenosylmethionine decarboxylase. Antimicrob Agents Chemother. 1990 Aug;34(8):1485–1490. doi: 10.1128/aac.34.8.1485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bitonti A. J., Dumont J. A., McCann P. P. Characterization of Trypanosoma brucei brucei S-adenosyl-L-methionine decarboxylase and its inhibition by Berenil, pentamidine and methylglyoxal bis(guanylhydrazone). Biochem J. 1986 Aug 1;237(3):685–689. doi: 10.1042/bj2370685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  7. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  8. Dorhout B., Te Velde R. J., Ferwerda H., Kingma A. W., De Hoog E., Muskiet F. A. In vivo growth inhibition of L1210 leukemia by 4-amidinoindan-1-one 2'-amidinohydrazone (CGP 48664A), a new inhibitor of S-adenosylmethionine decarboxylase. Int J Cancer. 1995 Apr 10;61(2):214–217. doi: 10.1002/ijc.2910610212. [DOI] [PubMed] [Google Scholar]
  9. Hafner E. W., Tabor C. W., Tabor H. Mutants of Escherichia coli that do not contain 1,4-diaminobutane (putrescine) or spermidine. J Biol Chem. 1979 Dec 25;254(24):12419–12426. [PubMed] [Google Scholar]
  10. Heby O., Persson L. Molecular genetics of polyamine synthesis in eukaryotic cells. Trends Biochem Sci. 1990 Apr;15(4):153–158. doi: 10.1016/0968-0004(90)90216-x. [DOI] [PubMed] [Google Scholar]
  11. Heby O. Role of polyamines in the control of cell proliferation and differentiation. Differentiation. 1981;19(1):1–20. doi: 10.1111/j.1432-0436.1981.tb01123.x. [DOI] [PubMed] [Google Scholar]
  12. Henkle K. J., Liebau E., Müller S., Bergmann B., Walter R. D. Characterization and molecular cloning of a Cu/Zn superoxide dismutase from the human parasite Onchocerca volvulus. Infect Immun. 1991 Jun;59(6):2063–2069. doi: 10.1128/iai.59.6.2063-2069.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hill J. R., Morris D. R. Cell-specific translation of S-adenosylmethionine decarboxylase mRNA. Regulation by the 5' transcript leader. J Biol Chem. 1992 Oct 25;267(30):21886–21893. [PubMed] [Google Scholar]
  14. Hill J. R., Morris D. R. Cell-specific translational regulation of S-adenosylmethionine decarboxylase mRNA. Dependence on translation and coding capacity of the cis-acting upstream open reading frame. J Biol Chem. 1993 Jan 5;268(1):726–731. [PubMed] [Google Scholar]
  15. Huber M., Poulin R. Antiproliferative effect of spermine depletion by N-cyclohexyl-1,3-diaminopropane in human breast cancer cells. Cancer Res. 1995 Feb 15;55(4):934–943. [PubMed] [Google Scholar]
  16. Jänne J., Alhonen L., Leinonen P. Polyamines: from molecular biology to clinical applications. Ann Med. 1991 Aug;23(3):241–259. doi: 10.3109/07853899109148056. [DOI] [PubMed] [Google Scholar]
  17. Kameji T., Pegg A. E. Inhibition of translation of mRNAs for ornithine decarboxylase and S-adenosylmethionine decarboxylase by polyamines. J Biol Chem. 1987 Feb 25;262(6):2427–2430. [PubMed] [Google Scholar]
  18. Kashiwagi K., Taneja S. K., Liu T. Y., Tabor C. W., Tabor H. Spermidine biosynthesis in Saccharomyces cerevisiae. Biosynthesis and processing of a proenzyme form of S-adenosylmethionine decarboxylase. J Biol Chem. 1990 Dec 25;265(36):22321–22328. [PubMed] [Google Scholar]
  19. Krause M., Hirsh D. A trans-spliced leader sequence on actin mRNA in C. elegans. Cell. 1987 Jun 19;49(6):753–761. doi: 10.1016/0092-8674(87)90613-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mad Arif S. A., Taylor M. A., George L. A., Butler A. R., Burch L. R., Davies H. V., Stark M. J., Kumar A. Characterisation of the S-adenosylmethionine decarboxylase (SAMDC) gene of potato. Plant Mol Biol. 1994 Oct;26(1):327–338. doi: 10.1007/BF00039543. [DOI] [PubMed] [Google Scholar]
  21. Maric S. C., Crozat A., Louhimo J., Knuutila S., Jänne O. A. The human S-adenosylmethionine decarboxylase gene: nucleotide sequence of a pseudogene and chromosomal localization of the active gene (AMD1) and the pseudogene (AMD2). Cytogenet Cell Genet. 1995;70(3-4):195–199. doi: 10.1159/000134032. [DOI] [PubMed] [Google Scholar]
  22. Marić S. C., Crozat A., Jänne O. A. Structure and organization of the human S-adenosylmethionine decarboxylase gene. J Biol Chem. 1992 Sep 15;267(26):18915–18923. [PubMed] [Google Scholar]
  23. Markham G. D., Tabor C. W., Tabor H. S-adenosylmethionine decarboxylase of Escherichia coli. Studies on the covalently linked pyruvate required for activity. J Biol Chem. 1982 Oct 25;257(20):12063–12068. [PubMed] [Google Scholar]
  24. Martin M. E., Piette J., Yaniv M., Tang W. J., Folk W. R. Activation of the polyomavirus enhancer by a murine activator protein 1 (AP1) homolog and two contiguous proteins. Proc Natl Acad Sci U S A. 1988 Aug;85(16):5839–5843. doi: 10.1073/pnas.85.16.5839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Marton L. J., Pegg A. E. Polyamines as targets for therapeutic intervention. Annu Rev Pharmacol Toxicol. 1995;35:55–91. doi: 10.1146/annurev.pa.35.040195.000415. [DOI] [PubMed] [Google Scholar]
  26. Moshier J. A., Dosescu J., Skunca M., Luk G. D. Transformation of NIH/3T3 cells by ornithine decarboxylase overexpression. Cancer Res. 1993 Jun 1;53(11):2618–2622. [PubMed] [Google Scholar]
  27. Pajunen A., Crozat A., Jänne O. A., Ihalainen R., Laitinen P. H., Stanley B., Madhubala R., Pegg A. E. Structure and regulation of mammalian S-adenosylmethionine decarboxylase. J Biol Chem. 1988 Nov 15;263(32):17040–17049. [PubMed] [Google Scholar]
  28. Pegg A. E., Jones D. B., Secrist J. A., 3rd Effect of inhibitors of S-adenosylmethionine decarboxylase on polyamine content and growth of L1210 cells. Biochemistry. 1988 Mar 8;27(5):1408–1415. doi: 10.1021/bi00405a003. [DOI] [PubMed] [Google Scholar]
  29. Pegg A. E., McCann P. P. S-adenosylmethionine decarboxylase as an enzyme target for therapy. Pharmacol Ther. 1992 Dec;56(3):359–377. doi: 10.1016/0163-7258(92)90025-u. [DOI] [PubMed] [Google Scholar]
  30. Pegg A. E. Polyamine metabolism and its importance in neoplastic growth and a target for chemotherapy. Cancer Res. 1988 Feb 15;48(4):759–774. [PubMed] [Google Scholar]
  31. Pegg A. E. Recent advances in the biochemistry of polyamines in eukaryotes. Biochem J. 1986 Mar 1;234(2):249–262. doi: 10.1042/bj2340249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Persson L., Stjernborg L., Holm I., Heby O. Polyamine-mediated control of mammalian S-adenosyl-L-methionine decarboxylase expression: effects on the content and translational efficiency of the mRNA. Biochem Biophys Res Commun. 1989 May 15;160(3):1196–1202. doi: 10.1016/s0006-291x(89)80130-5. [DOI] [PubMed] [Google Scholar]
  33. Pulkka A., Ihalainen R., Suorsa A., Riviere M., Szpirer J., Pajunen A. Structures and chromosomal localizations of two rat genes encoding S-adenosylmethionine decarboxylase. Genomics. 1993 May;16(2):342–349. doi: 10.1006/geno.1993.1195. [DOI] [PubMed] [Google Scholar]
  34. Pulkka A., Keränen M. R., Salmela A., Salmikangas P., Ihalainen R., Pajunen A. Nucleotide sequence of rat S-adenosylmethionine decarboxylase cDNA. Comparison with an intronless rat pseudogene. Gene. 1990 Feb 14;86(2):193–199. doi: 10.1016/0378-1119(90)90279-z. [DOI] [PubMed] [Google Scholar]
  35. Rathaur S., Wittich R. M., Walter R. D. Ascaris suum and Onchocerca volvulus: S-adenosylmethionine decarboxylase. Exp Parasitol. 1988 Apr;65(2):277–281. doi: 10.1016/0014-4894(88)90132-4. [DOI] [PubMed] [Google Scholar]
  36. Regenass U., Mett H., Stanek J., Mueller M., Kramer D., Porter C. W. CGP 48664, a new S-adenosylmethionine decarboxylase inhibitor with broad spectrum antiproliferative and antitumor activity. Cancer Res. 1994 Jun 15;54(12):3210–3217. [PubMed] [Google Scholar]
  37. Rogers S., Wells R., Rechsteiner M. Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. Science. 1986 Oct 17;234(4774):364–368. doi: 10.1126/science.2876518. [DOI] [PubMed] [Google Scholar]
  38. Ruan H., Hill J. R., Fatemie-Nainie S., Morris D. R. Cell-specific translational regulation of S-adenosylmethionine decarboxylase mRNA. Influence of the structure of the 5' transcript leader on regulation by the upstream open reading frame. J Biol Chem. 1994 Jul 8;269(27):17905–17910. [PubMed] [Google Scholar]
  39. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Schägger H., von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. doi: 10.1016/0003-2697(87)90587-2. [DOI] [PubMed] [Google Scholar]
  41. Shantz L. M., Viswanath R., Pegg A. E. Role of the 5'-untranslated region of mRNA in the synthesis of S-adenosylmethionine decarboxylase and its regulation by spermine. Biochem J. 1994 Sep 15;302(Pt 3):765–772. doi: 10.1042/bj3020765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sharma V., Tekwani B. L., Saxena J. K., Gupta S., Katiyar J. C., Chatterjee R. K., Ghatak S., Shukla O. P. Polyamine metabolism in some helminth parasites. Exp Parasitol. 1991 Jan;72(1):15–23. doi: 10.1016/0014-4894(91)90116-e. [DOI] [PubMed] [Google Scholar]
  43. Stanley B. A., Pegg A. E. Amino acid residues necessary for putrescine stimulation of human S-adenosylmethionine decarboxylase proenzyme processing and catalytic activity. J Biol Chem. 1991 Oct 5;266(28):18502–18506. [PubMed] [Google Scholar]
  44. Stanley B. A., Pegg A. E., Holm I. Site of pyruvate formation and processing of mammalian S-adenosylmethionine decarboxylase proenzyme. J Biol Chem. 1989 Dec 15;264(35):21073–21079. [PubMed] [Google Scholar]
  45. Stanley B. A., Shantz L. M., Pegg A. E. Expression of mammalian S-adenosylmethionine decarboxylase in Escherichia coli. Determination of sites for putrescine activation of activity and processing. J Biol Chem. 1994 Mar 18;269(11):7901–7907. [PubMed] [Google Scholar]
  46. Stjernborg L., Heby O., Mamont P., Persson L. Polyamine-mediated regulation of S-adenosylmethionine decarboxylase expression in mammalian cells. Studies using 5'-([(Z)-4-amino-2-butenyl]methylamino)-5'-deoxyadenosine, a suicide inhibitor of the enzyme. Eur J Biochem. 1993 Jun 15;214(3):671–676. doi: 10.1111/j.1432-1033.1993.tb17967.x. [DOI] [PubMed] [Google Scholar]
  47. Suzuki T., Kashiwagi K., Igarashi K. Polyamine regulation of S-adenosylmethionine decarboxylase synthesis through the 5'-untranslated region of its mRNA. Biochem Biophys Res Commun. 1993 Apr 30;192(2):627–634. doi: 10.1006/bbrc.1993.1461. [DOI] [PubMed] [Google Scholar]
  48. Svensson F., Kockum I., Persson L. Diethylglyoxal bis(guanylhydrazone), a potent inhibitor of mammalian S-adenosylmethionine decarboxylase. Effects on cell proliferation and polyamine metabolism in L1210 leukemia cells. Mol Cell Biochem. 1993 Jul 21;124(2):141–147. doi: 10.1007/BF00929206. [DOI] [PubMed] [Google Scholar]
  49. Tabor C. W., Tabor H. Polyamines in microorganisms. Microbiol Rev. 1985 Mar;49(1):81–99. doi: 10.1128/mr.49.1.81-99.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Tabor C. W., Tabor H. Polyamines. Annu Rev Biochem. 1984;53:749–790. doi: 10.1146/annurev.bi.53.070184.003533. [DOI] [PubMed] [Google Scholar]
  51. Tabor C. W., Tabor H. The speEspeD operon of Escherichia coli. Formation and processing of a proenzyme form of S-adenosylmethionine decarboxylase. J Biol Chem. 1987 Nov 25;262(33):16037–16040. [PubMed] [Google Scholar]
  52. White M. W., Degnin C., Hill J., Morris D. R. Specific regulation by endogenous polyamines of translational initiation of S-adenosylmethionine decarboxylase mRNA in Swiss 3T3 fibroblasts. Biochem J. 1990 Jun 15;268(3):657–660. doi: 10.1042/bj2680657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wittich R. M., Kilian H. D., Walter R. D. Polyamine metabolism in filarial worms. Mol Biochem Parasitol. 1987 Jun;24(2):155–162. doi: 10.1016/0166-6851(87)90102-2. [DOI] [PubMed] [Google Scholar]
  54. Wright P. S., Byers T. L., Cross-Doersen D. E., McCann P. P., Bitonti A. J. Irreversible inhibition of S-adenosylmethionine decarboxylase in Plasmodium falciparum-infected erythrocytes: growth inhibition in vitro. Biochem Pharmacol. 1991 Jun 1;41(11):1713–1718. doi: 10.1016/0006-2952(91)90174-4. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES