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. 1992 Mar 11;20(5):1123–1127. doi: 10.1093/nar/20.5.1123

Removal of a 3' non-coding sequence is an initial step in degradation of gro alpha mRNA and is regulated by interleukin-1.

M Y Stoeckle 1
PMCID: PMC312101  PMID: 1549476

Abstract

I have previously shown that IL-1 regulates the stability of gro alpha mRNA in fibroblasts and that decay is associated with appearance of a smaller species of gro RNA that lacks poly(A). In this study, the relationship between the two species of gro RNA, which migrate at 1.3 and 0.9 kilobases, was characterized. Following withdrawal of IL-1 or addition of IL-1 receptor antagonist, 1.3 kilobase gro alpha mRNA was rapidly degraded and this was associated with increased expression of the 0.9 kilobase RNA. This increase occurred in the presence of actinomycin D, indicating that the 0.9 kilobase gro RNA was a product of a pre-existing transcript. In cells treated with 1 pg/ml IL-1, both species were induced but the 0.9 kilobase RNA appeared later, consistent with a precursor-product relationship. In cells treated with higher doses of IL-1, the 0.9 kilobase RNA was not expressed. Using an RNAase protection assay, the 0.9 kilobase poly(A)-minus gro RNA was found to be derived from gro alpha mRNA by removal of a 130-nucleotide sequence from the 3' non-coding region. This is one of few examples of formation of an mRNA decay intermediate in vivo; it indicates that degradation of the body of gro alpha mRNA is initiated by site-specific nuclease attack. Characterization of the mechanism of gro alpha mRNA degradation is a first step towards identification of the ribonuclease that controls gro alpha mRNA stability.

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  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. Akashi M., Loussararian A. H., Adelman D. C., Saito M., Koeffler H. P. Role of lymphotoxin in expression of interleukin 6 in human fibroblasts. Stimulation and regulation. J Clin Invest. 1990 Jan;85(1):121–129. doi: 10.1172/JCI114401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anisowicz A., Bardwell L., Sager R. Constitutive overexpression of a growth-regulated gene in transformed Chinese hamster and human cells. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7188–7192. doi: 10.1073/pnas.84.20.7188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Anisowicz A., Zajchowski D., Stenman G., Sager R. Functional diversity of gro gene expression in human fibroblasts and mammary epithelial cells. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9645–9649. doi: 10.1073/pnas.85.24.9645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bandyopadhyay R., Coutts M., Krowczynska A., Brawerman G. Nuclease activity associated with mammalian mRNA in its native state: possible basis for selectivity in mRNA decay. Mol Cell Biol. 1990 May;10(5):2060–2069. doi: 10.1128/mcb.10.5.2060. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. 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]
  9. 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]
  10. Carter D. B., Deibel M. R., Jr, Dunn C. J., Tomich C. S., Laborde A. L., Slightom J. L., Berger A. E., Bienkowski M. J., Sun F. F., McEwan R. N. Purification, cloning, expression and biological characterization of an interleukin-1 receptor antagonist protein. Nature. 1990 Apr 12;344(6267):633–638. doi: 10.1038/344633a0. [DOI] [PubMed] [Google Scholar]
  11. Case J. P., Lafyatis R., Kumkumian G. K., Remmers E. F., Wilder R. L. IL-1 regulation of transin/stromelysin transcription in rheumatoid synovial fibroblasts appears to involve two antagonistic transduction pathways, an inhibitory, prostaglandin-dependent pathway mediated by cAMP, and a stimulatory, protein kinase C-dependent pathway. J Immunol. 1990 Dec 1;145(11):3755–3761. [PubMed] [Google Scholar]
  12. Dinarello C. A. Biology of interleukin 1. FASEB J. 1988 Feb;2(2):108–115. [PubMed] [Google Scholar]
  13. Eisenberg S. P., Evans R. J., Arend W. P., Verderber E., Brewer M. T., Hannum C. H., Thompson R. C. Primary structure and functional expression from complementary DNA of a human interleukin-1 receptor antagonist. Nature. 1990 Jan 25;343(6256):341–346. doi: 10.1038/343341a0. [DOI] [PubMed] [Google Scholar]
  14. Elias J. A., Lentz V. IL-1 and tumor necrosis factor synergistically stimulate fibroblast IL-6 production and stabilize IL-6 messenger RNA. J Immunol. 1990 Jul 1;145(1):161–166. [PubMed] [Google Scholar]
  15. Elias J. A., Reynolds M. M., Kotloff R. M., Kern J. A. Fibroblast interleukin 1 beta: synergistic stimulation by recombinant interleukin 1 and tumor necrosis factor and posttranscriptional regulation. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6171–6175. doi: 10.1073/pnas.86.16.6171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Golds E. E., Mason P., Nyirkos P. Inflammatory cytokines induce synthesis and secretion of gro protein and a neutrophil chemotactic factor but not beta 2-microglobulin in human synovial cells and fibroblasts. Biochem J. 1989 Apr 15;259(2):585–588. doi: 10.1042/bj2590585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hall D. J., Brownlee C., Stiles C. D. Interleukin-1 is a potent regulator of JE and KC gene expression in quiescent BALB/c fibroblasts. J Cell Physiol. 1989 Oct;141(1):154–159. doi: 10.1002/jcp.1041410123. [DOI] [PubMed] [Google Scholar]
  18. Haskill S., Peace A., Morris J., Sporn S. A., Anisowicz A., Lee S. W., Smith T., Martin G., Ralph P., Sager R. Identification of three related human GRO genes encoding cytokine functions. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7732–7736. doi: 10.1073/pnas.87.19.7732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jones T. R., Cole M. D. Rapid cytoplasmic turnover of c-myc mRNA: requirement of the 3' untranslated sequences. Mol Cell Biol. 1987 Dec;7(12):4513–4521. doi: 10.1128/mcb.7.12.4513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kowalski J., Denhardt D. T. Regulation of the mRNA for monocyte-derived neutrophil-activating peptide in differentiating HL60 promyelocytes. Mol Cell Biol. 1989 May;9(5):1946–1957. doi: 10.1128/mcb.9.5.1946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Laird-Offringa I. A., de Wit C. L., Elfferich P., van der Eb A. J. Poly(A) tail shortening is the translation-dependent step in c-myc mRNA degradation. Mol Cell Biol. 1990 Dec;10(12):6132–6140. doi: 10.1128/mcb.10.12.6132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lieberman A. P., Pitha P. M., Shin M. L. Protein kinase regulates tumor necrosis factor mRNA stability in virus-stimulated astrocytes. J Exp Med. 1990 Sep 1;172(3):989–992. doi: 10.1084/jem.172.3.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lindholm D., Heumann R., Hengerer B., Thoenen H. Interleukin 1 increases stability and transcription of mRNA encoding nerve growth factor in cultured rat fibroblasts. J Biol Chem. 1988 Nov 5;263(31):16348–16351. [PubMed] [Google Scholar]
  24. 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]
  25. Mizel S. B. How does interleukin 1 activate cells? Cyclic AMP and interleukin 1 signal transduction. Immunol Today. 1990 Nov;11(11):390–391. doi: 10.1016/0167-5699(90)90154-2. [DOI] [PubMed] [Google Scholar]
  26. Moser B., Clark-Lewis I., Zwahlen R., Baggiolini M. Neutrophil-activating properties of the melanoma growth-stimulatory activity. J Exp Med. 1990 May 1;171(5):1797–1802. doi: 10.1084/jem.171.5.1797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. O'Neill L. A., Bird T. A., Saklatvala J. How does interleukin 1 activate cells? Interleukin 1 signal transduction. Immunol Today. 1990 Nov;11(11):392–394. doi: 10.1016/0167-5699(90)90155-3. [DOI] [PubMed] [Google Scholar]
  28. Richmond A., Balentien E., Thomas H. G., Flaggs G., Barton D. E., Spiess J., Bordoni R., Francke U., Derynck R. Molecular characterization and chromosomal mapping of melanoma growth stimulatory activity, a growth factor structurally related to beta-thromboglobulin. EMBO J. 1988 Jul;7(7):2025–2033. doi: 10.1002/j.1460-2075.1988.tb03042.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Richmond A., Thomas H. G. Melanoma growth stimulatory activity: isolation from human melanoma tumors and characterization of tissue distribution. J Cell Biochem. 1988 Feb;36(2):185–198. doi: 10.1002/jcb.240360209. [DOI] [PubMed] [Google Scholar]
  30. Schröder J. M., Persoon N. L., Christophers E. Lipopolysaccharide-stimulated human monocytes secrete, apart from neutrophil-activating peptide 1/interleukin 8, a second neutrophil-activating protein. NH2-terminal amino acid sequence identity with melanoma growth stimulatory activity. J Exp Med. 1990 Apr 1;171(4):1091–1100. doi: 10.1084/jem.171.4.1091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Shyu A. B., Belasco J. G., Greenberg M. E. Two distinct destabilizing elements in the c-fos message trigger deadenylation as a first step in rapid mRNA decay. Genes Dev. 1991 Feb;5(2):221–231. doi: 10.1101/gad.5.2.221. [DOI] [PubMed] [Google Scholar]
  33. Shyu A. B., Greenberg M. E., Belasco J. G. The c-fos transcript is targeted for rapid decay by two distinct mRNA degradation pathways. Genes Dev. 1989 Jan;3(1):60–72. doi: 10.1101/gad.3.1.60. [DOI] [PubMed] [Google Scholar]
  34. Sims J. E., Acres R. B., Grubin C. E., McMahan C. J., Wignall J. M., March C. J., Dower S. K. Cloning the interleukin 1 receptor from human T cells. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8946–8950. doi: 10.1073/pnas.86.22.8946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Stoeckle M. Y., Barker K. A. Two burgeoning families of platelet factor 4-related proteins: mediators of the inflammatory response. New Biol. 1990 Apr;2(4):313–323. [PubMed] [Google Scholar]
  36. Stoeckle M. Y., Hanafusa H. Processing of 9E3 mRNA and regulation of its stability in normal and Rous sarcoma virus-transformed cells. Mol Cell Biol. 1989 Nov;9(11):4738–4745. doi: 10.1128/mcb.9.11.4738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Stoeckle M. Y. Post-transcriptional regulation of gro alpha, beta, gamma, and IL-8 mRNAs by IL-1 beta. Nucleic Acids Res. 1991 Feb 25;19(4):917–920. doi: 10.1093/nar/19.4.917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sussman M. Model for quantitative and qualitative control of mRNA translation in eukaryotes. Nature. 1970 Mar 28;225(5239):1245–1246. doi: 10.1038/2251245a0. [DOI] [PubMed] [Google Scholar]
  39. Swartwout S. G., Kinniburgh A. J. c-myc RNA degradation in growing and differentiating cells: possible alternate pathways. Mol Cell Biol. 1989 Jan;9(1):288–295. doi: 10.1128/mcb.9.1.288. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tekamp-Olson P., Gallegos C., Bauer D., McClain J., Sherry B., Fabre M., van Deventer S., Cerami A. Cloning and characterization of cDNAs for murine macrophage inflammatory protein 2 and its human homologues. J Exp Med. 1990 Sep 1;172(3):911–919. doi: 10.1084/jem.172.3.911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Thorens B., Mermod J. J., Vassalli P. Phagocytosis and inflammatory stimuli induce GM-CSF mRNA in macrophages through posttranscriptional regulation. Cell. 1987 Feb 27;48(4):671–679. doi: 10.1016/0092-8674(87)90245-5. [DOI] [PubMed] [Google Scholar]
  42. Tso J. Y., Sun X. H., Kao T. H., Reece K. S., Wu R. Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. Nucleic Acids Res. 1985 Apr 11;13(7):2485–2502. doi: 10.1093/nar/13.7.2485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. 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]
  44. Wen D. Z., Rowland A., Derynck R. Expression and secretion of gro/MGSA by stimulated human endothelial cells. EMBO J. 1989 Jun;8(6):1761–1766. doi: 10.1002/j.1460-2075.1989.tb03569.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Whittemore L. A., Maniatis T. Postinduction turnoff of beta-interferon gene expression. Mol Cell Biol. 1990 Apr;10(4):1329–1337. doi: 10.1128/mcb.10.4.1329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wilson T., Treisman R. Removal of poly(A) and consequent degradation of c-fos mRNA facilitated by 3' AU-rich sequences. Nature. 1988 Nov 24;336(6197):396–399. doi: 10.1038/336396a0. [DOI] [PubMed] [Google Scholar]
  47. Wodnar-Filipowicz A., Moroni C. Regulation of interleukin 3 mRNA expression in mast cells occurs at the posttranscriptional level and is mediated by calcium ions. Proc Natl Acad Sci U S A. 1990 Jan;87(2):777–781. doi: 10.1073/pnas.87.2.777. [DOI] [PMC free article] [PubMed] [Google Scholar]

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