Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1993 Apr 11;21(7):1613–1617. doi: 10.1093/nar/21.7.1613

High-resolution analysis of gro alpha mRNA poly(A) shortening: regulation by interleukin-1 beta.

M Y Stoeckle 1, L Guan 1
PMCID: PMC309371  PMID: 8097587

Abstract

We have previously shown that destabilization of gro alpha mRNA is associated with poly(A) shortening. In this study, we used high-resolution Northern blots to determine the rate and extent of gro alpha mRNA poly(A) shortening. gro alpha mRNA was found to undergo complete deadenylation within 2 h following withdrawal of IL-1. However, the process was not uniform: at 1 h following IL-1 withdrawal, gro alpha mRNA poly(A) lengths ranged from 0 to 180 nucleotides. There was an accumulation of deadenylated gro alpha mRNA which suggested that there may be another step before the mRNA is destroyed. Cycloheximide was found to block gro alpha mRNA degradation at the level of poly(A) shortening. Northern blots revealed a previously unrecognized periodic distribution of poly(A) lengths that was consistent with endonucleolytic cleavage between complexes of poly(A)-binding protein. The findings indicate that the degradation pathway of gro alpha mRNA is a slower version of the c-fos mRNA model, with the important additional feature that deadenylation and degradation are subject to physiologic regulation. This study provides a detailed picture of gro alpha mRNA poly(A) shortening and establishes a basis for further investigation of the mechanism by which IL-1 stabilizes specific mRNAs.

Full text

PDF
1613

Images in this article

1614Image
on p.1614
1614Image
on p.1614
1615Image
on p.1615
1615Image
on p.1615

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. 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]
  3. Anisowicz A., Messineo M., Lee S. W., Sager R. An NF-kappa B-like transcription factor mediates IL-1/TNF-alpha induction of gro in human fibroblasts. J Immunol. 1991 Jul 15;147(2):520–527. [PubMed] [Google Scholar]
  4. Baer B. W., Kornberg R. D. Repeating structure of cytoplasmic poly(A)-ribonucleoprotein. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1890–1892. doi: 10.1073/pnas.77.4.1890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Baer B. W., Kornberg R. D. The protein responsible for the repeating structure of cytoplasmic poly(A)-ribonucleoprotein. J Cell Biol. 1983 Mar;96(3):717–721. doi: 10.1083/jcb.96.3.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bernstein P., Ross J. Poly(A), poly(A) binding protein and the regulation of mRNA stability. Trends Biochem Sci. 1989 Sep;14(9):373–377. doi: 10.1016/0968-0004(89)90011-x. [DOI] [PubMed] [Google Scholar]
  7. Brawerman G. The Role of the poly(A) sequence in mammalian messenger RNA. CRC Crit Rev Biochem. 1981;10(1):1–38. doi: 10.3109/10409238109114634. [DOI] [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. 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]
  10. Peppel K., Vinci J. M., Baglioni C. The AU-rich sequences in the 3' untranslated region mediate the increased turnover of interferon mRNA induced by glucocorticoids. J Exp Med. 1991 Feb 1;173(2):349–355. doi: 10.1084/jem.173.2.349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Sachs A. B., Davis R. W. The poly(A) binding protein is required for poly(A) shortening and 60S ribosomal subunit-dependent translation initiation. Cell. 1989 Sep 8;58(5):857–867. doi: 10.1016/0092-8674(89)90938-0. [DOI] [PubMed] [Google Scholar]
  13. Sachs A. B., Deardorff J. A. Translation initiation requires the PAB-dependent poly(A) ribonuclease in yeast. Cell. 1992 Sep 18;70(6):961–973. doi: 10.1016/0092-8674(92)90246-9. [DOI] [PubMed] [Google Scholar]
  14. Sehgal P. B., Gupta S. L. Regulation of the stability of poly(I)xpoly(C)-induced human fibroblast interferon mRNA: selective inactivation of interferon mRNA and lack of involvement of 2',5'-oligo(A) synthetase activation during the shutoff of interferon production. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3489–3493. doi: 10.1073/pnas.77.6.3489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. 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]
  18. 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]
  19. Stoeckle M. Y. Removal of a 3' non-coding sequence is an initial step in degradation of gro alpha mRNA and is regulated by interleukin-1. Nucleic Acids Res. 1992 Mar 11;20(5):1123–1127. doi: 10.1093/nar/20.5.1123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. 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]

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

RESOURCES