Abstract
Oligodeoxynucleotides 18 nucleotides in length having sequences complementary to regions spanning the initiation codon regions of ornithine decarboyxlase or S-adenosylmethionine decarboxylase mRNAs were tested for their ability to inhibit translation of these mRNAs. In reticulocyte lysates, a strong and dose dependent reduction of ornithine decarboyxlase synthesis in response to mRNA from D-R L1210 cells was brought about by 5′-AAAGCT GCTCATGGTTCT-3′ which is complementary to the sequence from - 6 to + 12 of the mRNA sequence but there was no inhibition by 5′-TGCAGCTTCCATCACCGT-3′. Conversely, the latter oligodeoxynucleotide which is complementary to the sequence from − 6 to + 12 of the mRNA of S-adenosyl methionine decarboxylase was a strong inhibitor of the synthesis of this enzyme in response to rat prostate mRNA and the antisense sequence from ornithine decarboxylase had no effect. The translation of ornithine decarboxylase mRNA in a wheat germ system was inhibited by the antisense oligodeoxynucleotide at much lower concentration than those needed in the reticulocyte lysate suggesting that degradation of the hybrid by ribonuclease H may be an important factor in this inhibition. These results indicate that such oligonucleotides may be useful to regulate cellular polyamine levels and as probes to study control of mRNA translation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- ODC:
-
ornithine decarboxylase
- AdoMetDC:
-
S-adenosylmethionine decarboxylase
- DFMO:
-
difluoromethylornithine
References
Tabor CW, Tabor H: Polyamines. Annu Rev Biochem 53: 749–790, 1984
Pegg AE: Revent advances in biochemistry of polyamines in eukaryotes. Biochem J 234: 249–262, 1986
McCann PP, Pegg AE, Sjoerdsma A: In: PP McCann, AE Pegg, A Sjoerdsma (eds.) Inhibition of polyamine metabolism: biological significance and basis for new therapies. Academic Press, New York, 1987, pp 1–371
Pegg AE: Polyamine metabolism and its importance in neoplastic growth and a target for chemotherapy. Cancer Res 48: 525–542, 1986
Porter CW, Sufrin JR: Interference with polyamine biosynthesis and/or function by analog of polyamines of methionine as a potential anticancer chemotherapeutic strategy. Anticancer Res 6: 525–542, 1986
Bergeron RJ, Neims AH, McManis JS, Hawthorne TR, Vinson JRT, Bortell R, Ingeno MJ: Synthetic polyamine analogues as antineoplastics. J Med Chem 31: 1183–1190, 1988
Bergeron RJ, Hawthorne TR, Vinson JRT, Bechk DEI: Role of the methylene backbone in the antiproliferative activity of polyamine analogues on L1210 cells. Cancer Res 49: 2859–2964, 1989
Stein CA, Cohen JS: Oligodeoxynucleotides as inhibitors of gene expression: a review. Cancer Res 48: 2659–2668, 1988
Loose-Mitchell DS: Antisense nucleic acids as a potential class of pharmaceutical agents. TIPS 9: 45–47, 1988
Marcus-Sekura CJ: Techniques for using antisense oligodeoxyribonucleotides to study gene expression. Anal Biochem 172: 289–295, 1988
Pegg AE, Secrist JA, Madhubala R: Properties of L1210 cells resistant to α-diluoromethylornithine (DFMO). Cancer Res 48: 2678–2682, 1988
Shirahata A, Pegg AE: Regulation of s-adenosylmethionine decarboxylase activity in rat liver and prostate. J Biol Chem 260: 9583–9588, 1985
Kameji T, Pegg AE: Effect of putrescine on the synthesis of s-adenosyl methionine decarboxylase. Biochem J 243: 285–288, 1987
Kameji T, Pegg AE: Inhibition of translation of mRNA for ornithine decarboxylase and s-adenosyl methionine decarboxylase by polyamines. J Biol Chem 262: 2427–2430, 1987
Kahana C, Nathans D: Isolation of cloned cDNA encoding mammalian ornithine decarboxylase. Proc Natl Acad Sci USA 81: 3645–3649, 1984
Pajunen A, Crozat A, Janne OA, Ihalainen R, Laitinen PH, Stanley B, Madhubala R, Pegg AE: Structure and regulation of mammalian s-adenosylmethionine decarboxylase. J Biol Chem 263: 17040–17049, 1988
Blake KR, Murakami A, Miller PS: Inhibition of rabbit globin mRNA translation by sequence specific oligodeoxy ribonucleoside. Biochem 24: 6132–6138, 1985
Goodchild J, Carrol E, Greenberg JR: Inhibition of rabbit β-globin synthesis by complementary oligonucleotides: identification of mRNA sites sensitive to inhibition. Arch Biochem Biophys 263: 401–409, 1988
Walder RY, Walder JA: Role of RNase H in hybrid-arrested translation by antisense oligonucleotides. Proc Natl Acad Sci USA 85: 5011–5015, 1988
Loke SL, Stein CA, Zhang XH, Mori K, Nakanishi M, Subasinghe C, Cohen JS, Neckers LM: Characterization of oligonucleotide transport into living cells. Proc Natl Acad Sci USA 86: 3474–3478, 1989
Maher LJ, Dolnick BJ: Comparative hybrid arrest by tandem antisense oligodeoxyribonucleotides or oligodeoxy-ribonucleoside methylphosphonates in a cell-free system. Nucle Acids Res 16: 3341–3358, 1988
Eckstein F, Gish G: Phosphorothioates in molecular biology. TIBS 14: 97–100, 1989
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Madhubala, R., Pegg, A.E. Inhibition of ornithine decarboxylase and S-adenosylmethionine decarboxylase synthesis by antisense oligodeoxynucleotides. Mol Cell Biochem 116, 191–195 (1992). https://doi.org/10.1007/BF00299398
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00299398