Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1993 Jan 11;21(1):155–162. doi: 10.1093/nar/21.1.155

An RNasin-resistant ribonuclease selective for interleukin 2 mRNA.

J Hua 1, R Garner 1, V Paetkau 1
PMCID: PMC309078  PMID: 8441610

Abstract

Interleukin 2 (IL2) mRNA has a short half-life in the cytoplasm of T lymphocytes, relative to most mRNA. We have discovered a candidate ribonuclease to account for the rapid turnover of IL2 mRNA in the cytosol of the human T lymphocyte cell line Jurkat. In partially purified form, this RNase is about 7 times as active on IL2 as on beta-globin mRNA. Pancreatic RNase, by contrast, does not show a significant preference for IL2 mRNA. Neither 5' capping, nor polyadenylation of the substrate mRNAs affects their degradation by the IL2-selective mRNase, whose activity is optimal in 0.5 mM Mg++ and 100 mM potassium acetate. The mRNase behaves like a protein of molecular weight 60-70,000 on gel chromatography, and is unusual in that it is insensitive to placental RNase inhibitor (RNasin). The mRNase cleaves IL2 mRNA at a small number of sites in the coding region, and IL2 mRNA containing only the coding region and 36 nucleotides of the 3'-noncoding region competes efficiently with full-length IL2 mRNA for the mRNase, whereas beta-globin mRNA does not.

Full text

PDF
160

Images in this article

157Image
on p.157
159Image
on p.159
160Image
on p.160
161Image
on p.161

Selected References

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

  1. Akahane K., Cohen R. B., Bickel M., Pluznik D. H. IL-1 alpha induces granulocyte-macrophage colony-stimulating factor gene expression in murine B lymphocyte cell lines via mRNA stabilization. J Immunol. 1991 Jun 15;146(12):4190–4196. [PubMed] [Google Scholar]
  2. Bernstein P., Peltz S. W., Ross J. The poly(A)-poly(A)-binding protein complex is a major determinant of mRNA stability in vitro. Mol Cell Biol. 1989 Feb;9(2):659–670. doi: 10.1128/mcb.9.2.659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beutler B., Thompson P., Keyes J., Hagerty K., Crawford D. Assay of a ribonuclease that preferentially hydrolyses mRNAs containing cytokine-derived UA-rich instability sequences. Biochem Biophys Res Commun. 1988 May 16;152(3):973–980. doi: 10.1016/s0006-291x(88)80379-6. [DOI] [PubMed] [Google Scholar]
  4. Bickel M., Cohen R. B., Pluznik D. H. Post-transcriptional regulation of granulocyte-macrophage colony-stimulating factor synthesis in murine T cells. J Immunol. 1990 Aug 1;145(3):840–845. [PubMed] [Google Scholar]
  5. Blobel G., Potter V. R. Relation of ribonuclease and ribonuclease inhibitor to the isolation of polysomes from rat liver. Proc Natl Acad Sci U S A. 1966 May;55(5):1283–1288. doi: 10.1073/pnas.55.5.1283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bohjanen P. R., Petryniak B., June C. H., Thompson C. B., Lindsten T. An inducible cytoplasmic factor (AU-B) binds selectively to AUUUA multimers in the 3' untranslated region of lymphokine mRNA. Mol Cell Biol. 1991 Jun;11(6):3288–3295. doi: 10.1128/mcb.11.6.3288. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brewer G. An A + U-rich element RNA-binding factor regulates c-myc mRNA stability in vitro. Mol Cell Biol. 1991 May;11(5):2460–2466. doi: 10.1128/mcb.11.5.2460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brewer G., Ross J. Poly(A) shortening and degradation of the 3' A+U-rich sequences of human c-myc mRNA in a cell-free system. Mol Cell Biol. 1988 Apr;8(4):1697–1708. doi: 10.1128/mcb.8.4.1697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Brewer G., Ross J. Regulation of c-myc mRNA stability in vitro by a labile destabilizer with an essential nucleic acid component. Mol Cell Biol. 1989 May;9(5):1996–2006. doi: 10.1128/mcb.9.5.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Caput D., Beutler B., Hartog K., Thayer R., Brown-Shimer S., Cerami A. Identification of a common nucleotide sequence in the 3'-untranslated region of mRNA molecules specifying inflammatory mediators. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1670–1674. doi: 10.1073/pnas.83.6.1670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dani C., Mechti N., Piechaczyk M., Lebleu B., Jeanteur P., Blanchard J. M. Increased rate of degradation of c-myc mRNA in interferon-treated Daudi cells. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4896–4899. doi: 10.1073/pnas.82.15.4896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Greenberg J. R. High stability of messenger RNA in growing cultured cells. Nature. 1972 Nov 10;240(5376):102–104. doi: 10.1038/240102a0. [DOI] [PubMed] [Google Scholar]
  13. Hargrove J. L., Schmidt F. H. The role of mRNA and protein stability in gene expression. FASEB J. 1989 Oct;3(12):2360–2370. doi: 10.1096/fasebj.3.12.2676679. [DOI] [PubMed] [Google Scholar]
  14. Harpold M. M., Wilson M. C., Darnell J. E., Jr Chinese hamster polyadenylated messenger ribonucleic acid: relationship to non-polyadenylated sequences and relative conservation during messenger ribonucleic acid processing. Mol Cell Biol. 1981 Feb;1(2):188–198. doi: 10.1128/mcb.1.2.188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. June C. H., Ledbetter J. A., Lindsten T., Thompson C. B. Evidence for the involvement of three distinct signals in the induction of IL-2 gene expression in human T lymphocytes. J Immunol. 1989 Jul 1;143(1):153–161. [PubMed] [Google Scholar]
  16. June C. H., Ledbetter J. A., Linsley P. S., Thompson C. B. Role of the CD28 receptor in T-cell activation. Immunol Today. 1990 Jun;11(6):211–216. doi: 10.1016/0167-5699(90)90085-n. [DOI] [PubMed] [Google Scholar]
  17. Lang K. M., Spritz R. A. In vitro splicing pathways of pre-mRNAs containing multiple intervening sequences? Mol Cell Biol. 1987 Oct;7(10):3428–3437. doi: 10.1128/mcb.7.10.3428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ledbetter J. A., Imboden J. B., Schieven G. L., Grosmaire L. S., Rabinovitch P. S., Lindsten T., Thompson C. B., June C. H. CD28 ligation in T-cell activation: evidence for two signal transduction pathways. Blood. 1990 Apr 1;75(7):1531–1539. [PubMed] [Google Scholar]
  19. Lindstein T., June C. H., Ledbetter J. A., Stella G., Thompson C. B. Regulation of lymphokine messenger RNA stability by a surface-mediated T cell activation pathway. Science. 1989 Apr 21;244(4902):339–343. doi: 10.1126/science.2540528. [DOI] [PubMed] [Google Scholar]
  20. Pei R., Calame K. Differential stability of c-myc mRNAS in a cell-free system. Mol Cell Biol. 1988 Jul;8(7):2860–2868. doi: 10.1128/mcb.8.7.2860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rodgers J. R., Johnson M. L., Rosen J. M. Measurement of mRNA concentration and mRNA half-life as a function of hormonal treatment. Methods Enzymol. 1985;109:572–592. doi: 10.1016/0076-6879(85)09116-9. [DOI] [PubMed] [Google Scholar]
  22. Ross J., Kobs G., Brewer G., Peltz S. W. Properties of the exonuclease activity that degrades H4 histone mRNA. J Biol Chem. 1987 Jul 5;262(19):9374–9381. [PubMed] [Google Scholar]
  23. Shaw G., Kamen R. A conserved AU sequence from the 3' untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell. 1986 Aug 29;46(5):659–667. doi: 10.1016/0092-8674(86)90341-7. [DOI] [PubMed] [Google Scholar]
  24. Shaw J., Meerovitch K., Bleackley R. C., Paetkau V. Mechanisms regulating the level of IL-2 mRNA in T lymphocytes. J Immunol. 1988 Apr 1;140(7):2243–2248. [PubMed] [Google Scholar]
  25. Shaw J., Meerovitch K., Elliott J. F., Bleackley R. C., Paetkau V. Induction, suppression and superinduction of lymphokine mRNA in T lymphocytes. Mol Immunol. 1987 May;24(5):409–419. doi: 10.1016/0161-5890(87)90014-9. [DOI] [PubMed] [Google Scholar]
  26. Volloch V., Housman D. Stability of globin mRNA in terminally differentiating murine erythroleukemia cells. Cell. 1981 Feb;23(2):509–514. doi: 10.1016/0092-8674(81)90146-x. [DOI] [PubMed] [Google Scholar]
  27. Wager R. E., Assoian R. K. A phorbol ester-regulated ribonuclease system controlling transforming growth factor beta 1 gene expression in hematopoietic cells. Mol Cell Biol. 1990 Nov;10(11):5983–5990. doi: 10.1128/mcb.10.11.5983. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES