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PMID: 16954375 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Destabilization of interleukin-6 mRNA requires a putative RNA stem-loop structure, an AU-rich element, and the RNA-binding protein AUF1.

Molecular and cellular biology ·Vol. 26 ·No. 22 ·2006-11-00 ·Pages 8228-41

Paschoud S, Dogar AM, Kuntz C, Grisoni-Neupert B, Richman L, Kühn LC

Abstract

Interleukin-6 mRNA is unstable and degraded with a half-life of 30 min. Instability determinants can entirely be attributed to the 3' untranslated region. By grafting segments of this region to stable green fluorescent protein mRNA and subsequent scanning mutagenesis, we have identified two conserved elements, which together account for most of the instability. The first corresponds to a short noncanonical AU-rich element. The other, 80 nucleotides further 5', comprises a sequence predicted to form a stem-loop structure. Neither element alone was sufficient to confer full instability, suggesting that they might cooperate. Overexpression of myc-tagged AUF1 p37 and p42 isoforms as well as suppression of endogenous AUF1 by RNA interference stabilized interleukin-6 mRNA. Both effects required the AU-rich instability element. Similarly, the proteasome inhibitor MG132 stabilized interleukin-6 mRNA probably through an increase of AUF1 levels. The mRNA coimmunoprecipitated specifically with myc-tagged AUF1 p37 and p42 in cell extracts but only when the AU-rich instability element was present. These results indicate that AUF1 binds to the AU-rich element in vivo and promotes IL-6 mRNA degradation.

MeSH Terms
3' Untranslated Regions Animals Base Sequence COS Cells Chlorocebus aethiops Green Fluorescent Proteins/genetics Heterogeneous Nuclear Ribonucleoprotein D0 Heterogeneous-Nuclear Ribonucleoprotein D/genetics,metabolism Interleukin-6/genetics,metabolism Molecular Sequence Data Mutagenesis Point Mutation Proteasome Endopeptidase Complex/genetics RNA/metabolism RNA Processing, Post-Transcriptional RNA Stability Sequence Homology, Nucleic Acid
Chemicals
3' Untranslated Regions Heterogeneous Nuclear Ribonucleoprotein D0 Heterogeneous-Nuclear Ribonucleoprotein D Interleukin-6 Green Fluorescent Proteins RNA Proteasome Endopeptidase Complex
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Paschoud Serge
Swiss Institute for Experimental Cancer Research, Genetics Unit, Chemin des Boveresses 155, CH-1066 Epalinges, Switzerland. [email protected].
Dogar Afzal M
Kuntz Catherine
Grisoni-Neupert Barbara
Richman Larry
Kühn Lukas C
References (72)
72 references, click to expand
  1. SV40-transformed simian cells support the replication of early SV40 mutants.
    Cell. 1981 Jan;23(1):175-82 PMID: 6260373
  2. Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene.
    Nature. 1984 Oct 4-10;311(5985):433-8 PMID: 6090941
  3. Removal of a 67-base-pair sequence in the noncoding region of protooncogene fos converts it to a transforming gene.
    Proc Natl Acad Sci U S A. 1985 Aug;82(15):4987-91 PMID: 2991903
  4. Transient accumulation of c-fos RNA following serum stimulation requires a conserved 5' element and c-fos 3' sequences.
    Cell. 1985 Oct;42(3):889-902 PMID: 2414012
  5. A conserved AU sequence from the 3' untranslated region of GM-CSF mRNA mediates selective mRNA degradation.
    Cell. 1986 Aug 29;46(5):659-67 PMID: 3488815
  6. Noncoding 3' sequences of the transferrin receptor gene are required for mRNA regulation by iron.
    EMBO J. 1987 May;6(5):1287-93 PMID: 3608980
  7. Removal of poly(A) and consequent degradation of c-fos mRNA facilitated by 3' AU-rich sequences.
    Nature. 1988 Nov 24;336(6197):396-9 PMID: 3194021
  8. The mouse homologue of the human acidic ribosomal phosphoprotein PO: a highly conserved polypeptide that is under translational control.
    Nucleic Acids Res. 1989 Aug 11;17(15):6408 PMID: 2771657
  9. The nonamer UUAUUUAUU is the key AU-rich sequence motif that mediates mRNA degradation.
    Mol Cell Biol. 1995 Apr;15(4):2219-30 PMID: 7891716
  10. Interleukin 6 is essential for in vivo development of B lineage neoplasms.
    J Exp Med. 1995 Jul 1;182(1):243-8 PMID: 7790819
  11. Liver failure and defective hepatocyte regeneration in interleukin-6-deficient mice.
    Science. 1996 Nov 22;274(5291):1379-83 PMID: 8910279
  12. A cytokine mRNA-destabilizing element that is structurally and functionally distinct from A+U-rich elements.
    Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13721-5 PMID: 8943001
  13. Modulation of the fate of cytoplasmic mRNA by AU-rich elements: key sequence features controlling mRNA deadenylation and decay.
    Mol Cell Biol. 1997 Aug;17(8):4611-21 PMID: 9234718
  14. Structure and dynamics of the iron responsive element RNA: implications for binding of the RNA by iron regulatory binding proteins.
    J Mol Biol. 1997 Nov 21;274(1):72-83 PMID: 9398517
  15. Structure and genomic organization of the human AUF1 gene: alternative pre-mRNA splicing generates four protein isoforms.
    Genomics. 1998 Mar 1;48(2):195-202 PMID: 9521873
  16. Overexpression of HuR, a nuclear-cytoplasmic shuttling protein, increases the in vivo stability of ARE-containing mRNAs.
    EMBO J. 1998 Jun 15;17(12):3448-60 PMID: 9628880
  17. Assembly of AUF1 oligomers on U-rich RNA targets by sequential dimer association.
    J Biol Chem. 1999 Nov 19;274(47):33374-81 PMID: 10559216
  18. Somatic mRNA turnover mutants implicate tristetraprolin in the interleukin-3 mRNA degradation pathway.
    Mol Cell Biol. 2000 Jun;20(11):3753-63 PMID: 10805719
  19. A regulatory network for the efficient control of transgene expression.
    J Gene Med. 2000 Mar-Apr;2(2):107-16 PMID: 10809144
  20. A mechanism for translationally coupled mRNA turnover: interaction between the poly(A) tail and a c-fos RNA coding determinant via a protein complex.
    Cell. 2000 Sep 29;103(1):29-40 PMID: 11051545
  21. Versatile role for hnRNP D isoforms in the differential regulation of cytoplasmic mRNA turnover.
    Mol Cell Biol. 2001 Oct;21(20):6960-71 PMID: 11564879
  22. Genomic-scale measurement of mRNA turnover and the mechanisms of action of the anti-cancer drug flavopiridol.
    Genome Biol. 2001;2(10):RESEARCH0041 PMID: 11597333
  23. AU binding proteins recruit the exosome to degrade ARE-containing mRNAs.
    Cell. 2001 Nov 16;107(4):451-64 PMID: 11719186
  24. Cellular mutants define a common mRNA degradation pathway targeting cytokine AU-rich elements.
    RNA. 2001 Nov;7(11):1578-88 PMID: 11720287
  25. MK2 targets AU-rich elements and regulates biosynthesis of tumor necrosis factor and interleukin-6 independently at different post-transcriptional levels.
    J Biol Chem. 2002 Feb 1;277(5):3065-8 PMID: 11741878
  26. Ubiquitin-dependent mechanism regulates rapid turnover of AU-rich cytokine mRNAs.
    Proc Natl Acad Sci U S A. 2002 Feb 19;99(4):1842-6 PMID: 11842200
  27. RNA destabilization by the granulocyte colony-stimulating factor stem-loop destabilizing element involves a single stem-loop that promotes deadenylation.
    Mol Cell Biol. 2002 Mar;22(6):1664-73 PMID: 11865046
  28. A new player in oncogenesis: AUF1/hnRNPD overexpression leads to tumorigenesis in transgenic mice.
    Cancer Res. 2002 Mar 1;62(5):1489-95 PMID: 11888925
  29. Alternate exon insertion controls selective ubiquitination and degradation of different AUF1 protein isoforms.
    Nucleic Acids Res. 2002 Jul 15;30(14):3052-8 PMID: 12136087
  30. Functional cloning of BRF1, a regulator of ARE-dependent mRNA turnover.
    EMBO J. 2002 Sep 2;21(17):4709-18 PMID: 12198173
  31. IL-6 in autoimmune disease and chronic inflammatory proliferative disease.
    Cytokine Growth Factor Rev. 2002 Aug-Oct;13(4-5):357-68 PMID: 12220549
  32. Stable suppression of tumorigenicity by virus-mediated RNA interference.
    Cancer Cell. 2002 Sep;2(3):243-7 PMID: 12242156
  33. Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes.
    Nucleic Acids Res. 2002 Dec 15;30(24):5529-38 PMID: 12490721
  34. A constitutive decay element promotes tumor necrosis factor alpha mRNA degradation via an AU-rich element-independent pathway.
    Mol Cell Biol. 2003 May;23(10):3506-15 PMID: 12724409
  35. Tristetraprolin and its family members can promote the cell-free deadenylation of AU-rich element-containing mRNAs by poly(A) ribonuclease.
    Mol Cell Biol. 2003 Jun;23(11):3798-812 PMID: 12748283
  36. Mfold web server for nucleic acid folding and hybridization prediction.
    Nucleic Acids Res. 2003 Jul 1;31(13):3406-15 PMID: 12824337
  37. Decay rates of human mRNAs: correlation with functional characteristics and sequence attributes.
    Genome Res. 2003 Aug;13(8):1863-72 PMID: 12902380
  38. Selective degradation of AU-rich mRNAs promoted by the p37 AUF1 protein isoform.
    Mol Cell Biol. 2003 Sep;23(18):6685-93 PMID: 12944492
  39. Interleukin-6, CD45 and the src-kinases in myeloma cell proliferation.
    Leuk Lymphoma. 2003 Sep;44(9):1477-81 PMID: 14565647
  40. Targeted anti-interleukin-6 monoclonal antibody therapy for cancer: a review of the rationale and clinical evidence.
    Clin Cancer Res. 2003 Oct 15;9(13):4653-65 PMID: 14581334
  41. Human PABP binds AU-rich RNA via RNA-binding domains 3 and 4.
    Eur J Biochem. 2004 Jan;271(2):450-7 PMID: 14717712
  42. Roles of AUF1 isoforms, HuR and BRF1 in ARE-dependent mRNA turnover studied by RNA interference.
    Nucleic Acids Res. 2004;32(4):1279-88 PMID: 14976220
  43. Role of the 3'-untranslated region of human endothelin-1 in vascular endothelial cells. Contribution to transcript lability and the cellular heat shock response.
    J Biol Chem. 2004 Mar 5;279(10):8655-67 PMID: 14660616
  44. Functional dissection of hnRNP D suggests that nuclear import is required before hnRNP D can modulate mRNA turnover in the cytoplasm.
    RNA. 2004 Apr;10(4):669-80 PMID: 15037776
  45. Distinct domains of AU-rich elements exert different functions in mRNA destabilization and stabilization by p38 mitogen-activated protein kinase or HuR.
    Mol Cell Biol. 2004 Jun;24(11):4835-47 PMID: 15143177
  46. A KH domain RNA binding protein, KSRP, promotes ARE-directed mRNA turnover by recruiting the degradation machinery.
    Mol Cell. 2004 Jun 4;14(5):571-83 PMID: 15175153
  47. 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):161-6 PMID: 2358670
  48. Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line.
    Nucleic Acids Res. 1990 Jun 25;18(12):3587-96 PMID: 2194165
  49. RNA stabilization by the AU-rich element binding protein, HuR, an ELAV protein.
    EMBO J. 1998 Jun 15;17(12):3461-70 PMID: 9628881
  50. Feedback inhibition of macrophage tumor necrosis factor-alpha production by tristetraprolin.
    Science. 1998 Aug 14;281(5379):1001-5 PMID: 9703499
  51. Regulation of interleukin-1beta-induced interleukin-6 gene expression in human fibroblast-like synoviocytes by p38 mitogen-activated protein kinase.
    J Biol Chem. 1998 Sep 18;273(38):24832-8 PMID: 9733787
  52. Adenovirus-mediated regulable target gene expression in vivo.
    Proc Natl Acad Sci U S A. 1999 Jan 19;96(2):355-60 PMID: 9892637
  53. Control of mRNA decay by heat shock-ubiquitin-proteasome pathway.
    Science. 1999 Apr 16;284(5413):499-502 PMID: 10205060
  54. Unraveling a cytoplasmic role for hnRNP D in the in vivo mRNA destabilization directed by the AU-rich element.
    Genes Dev. 1999 Jul 15;13(14):1884-97 PMID: 10421639
  55. The p38 MAP kinase pathway signals for cytokine-induced mRNA stabilization via MAP kinase-activated protein kinase 2 and an AU-rich region-targeted mechanism.
    EMBO J. 1999 Sep 15;18(18):4969-80 PMID: 10487749
  56. Involvement of microRNA in AU-rich element-mediated mRNA instability.
    Cell. 2005 Mar 11;120(5):623-34 PMID: 15766526
  57. Multiple processing body factors and the ARE binding protein TTP activate mRNA decapping.
    Mol Cell. 2005 Dec 22;20(6):905-15 PMID: 16364915
  58. ARE-mRNA degradation requires the 5'-3' decay pathway.
    EMBO Rep. 2006 Jan;7(1):72-7 PMID: 16299471
  59. Assembly of AUF1 with eIF4G-poly(A) binding protein complex suggests a translation function in AU-rich mRNA decay.
    RNA. 2006 May;12(5):883-93 PMID: 16556936
  60. The protein-coding region of c-myc mRNA contains a sequence that specifies rapid mRNA turnover and induction by protein synthesis inhibitors.
    Genes Dev. 1991 Feb;5(2):232-43 PMID: 1995415
  61. An A + U-rich element RNA-binding factor regulates c-myc mRNA stability in vitro.
    Mol Cell Biol. 1991 May;11(5):2460-6 PMID: 1901943
  62. Generation of plasmacytomas with the chromosomal translocation t(12;15) in interleukin 6 transgenic mice.
    Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):232-5 PMID: 1729694
  63. Deadenylylation: a mechanism controlling c-fos mRNA decay.
    Enzyme. 1990;44(1-4):181-92 PMID: 2133650
  64. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters.
    Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5547-51 PMID: 1319065
  65. Strong conservation of non-coding sequences during vertebrates evolution: potential involvement in post-transcriptional regulation of gene expression.
    Nucleic Acids Res. 1993 May 25;21(10):2315-22 PMID: 8506129
  66. Purification, characterization, and cDNA cloning of an AU-rich element RNA-binding protein, AUF1.
    Mol Cell Biol. 1993 Dec;13(12):7652-65 PMID: 8246982
  67. The half-life of c-myc mRNA in growing and serum-stimulated cells: influence of the coding and 3' untranslated regions and role of ribosome translocation.
    Mol Cell Biol. 1994 Mar;14(3):2119-28 PMID: 8114742
  68. Optimal sequence and structure of iron-responsive elements. Selection of RNA stem-loops with high affinity for iron regulatory factor.
    J Biol Chem. 1994 Jul 1;269(26):17481-9 PMID: 8021254
  69. AUUUA is not sufficient to promote poly(A) shortening and degradation of an mRNA: the functional sequence within AU-rich elements may be UUAUUUA(U/A)(U/A).
    Mol Cell Biol. 1994 Dec;14(12):7984-95 PMID: 7969138
  70. Concurrent versus individual binding of HuR and AUF1 to common labile target mRNAs.
    EMBO J. 2004 Aug 4;23(15):3092-102 PMID: 15257295
  71. Liver regeneration: from myth to mechanism.
    Nat Rev Mol Cell Biol. 2004 Oct;5(10):836-47 PMID: 15459664
  72. A new technique for the assay of infectivity of human adenovirus 5 DNA.
    Virology. 1973 Apr;52(2):456-67 PMID: 4705382
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.01155-06
Email
[email protected]
Published
2006-11-00
Epub
2006-00-05
Pages
8228-41
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC1636780
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.01155-06 PMC Full Text View journal details
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