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PMID: 9214631 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

IkappaB alpha is a target for the mitogen-activated 90 kDa ribosomal S6 kinase.

The EMBO journal ·Vol. 16 ·No. 11 ·1997-06-02 ·Pages 3133-44

Schouten GJ, Vertegaal AC, Whiteside ST, Israël A, Toebes M, Dorsman JC, van der Eb AJ, Zantema A

Abstract

The activity of transcription factor NFkappaB is regulated by its subcellular localization. In most cell types, NFkappaB is sequestered in the cytoplasm due to binding of the inhibitory protein IkappaB alpha. Stimulation of cells with a wide variety of agents results in degradation of IkappaB alpha which allows translocation of NFkappaB to the nucleus. Degradation of IkappaB alpha is triggered by phosphorylation of two serine residues, i.e. Ser32 and Ser36, by as yet unknown kinases. Here we report that the mitogen-activated 90 kDa ribosomal S6 kinase (p90rsk1) is an IkappaB alpha kinase. p90rsk1 phosphorylates IkappaB alpha at Ser32 and it physically associates with IkappaB alpha in vivo. Moreover, when the function of p90rsk1 is impaired by expression of a dominant-negative mutant, IkappaB alpha degradation in response to mitogenic stimuli, e.g. 12-O-tetradecanoylphorbol 13-acetate (TPA), is inhibited. Finally, NFkappaB cannot be activated by TPA in cell lines that have low levels of p90rsk1. We conclude that p90rsk1 is an essential kinase required for phosphorylation and subsequent degradation of IkappaB alpha in response to mitogens.

MeSH Terms
Animals COS Cells Calcium-Calmodulin-Dependent Protein Kinases/metabolism Cell Transformation, Neoplastic Cell Transformation, Viral DNA-Binding Proteins/metabolism Humans I-kappa B Proteins Mitogens/pharmacology Mutation NF-KappaB Inhibitor alpha NF-kappa B/metabolism NF-kappa B p50 Subunit Phosphorylation Precipitin Tests Protein Binding Protein Precursors/metabolism Protein Serine-Threonine Kinases/metabolism Ribosomal Protein S6 Kinases Serine/metabolism Signal Transduction Substrate Specificity Tetradecanoylphorbol Acetate/pharmacology
Chemicals
DNA-Binding Proteins I-kappa B Proteins Mitogens NF-kappa B NF-kappa B p50 Subunit NFKBIA protein, human Protein Precursors NF-KappaB Inhibitor alpha Serine Protein Serine-Threonine Kinases Ribosomal Protein S6 Kinases Calcium-Calmodulin-Dependent Protein Kinases Tetradecanoylphorbol Acetate
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Schouten G J
Laboratory for Molecular Carcinogenesis, Sylvius Laboratories, Leiden University, The Netherlands.
Vertegaal A C
Whiteside S T
Israël A
Toebes M
Dorsman J C
van der Eb A J
Zantema A
References (54)
54 references, click to expand
  1. Assay of transforming activity of tumor virus DNA.
    Methods Enzymol. 1980;65(1):826-39 PMID: 6246378
  2. Casein kinase II phosphorylates I kappa B alpha at S-283, S-289, S-293, and T-291 and is required for its degradation.
    Mol Cell Biol. 1996 Mar;16(3):899-906 PMID: 8622692
  3. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product.
    Mol Cell Biol. 1985 Dec;5(12):3610-6 PMID: 3915782
  4. Insulin-stimulated MAP-2 kinase phosphorylates and activates ribosomal protein S6 kinase II.
    Nature. 1988 Aug 25;334(6184):715-8 PMID: 2842685
  5. Substrate specificity of ribosomal protein S6 kinase II from Xenopus eggs.
    Second Messengers Phosphoproteins. 1988;12(2-3):135-43 PMID: 3244115
  6. Sequence and expression of chicken and mouse rsk: homologs of Xenopus laevis ribosomal S6 kinase.
    Mol Cell Biol. 1989 Sep;9(9):3850-9 PMID: 2779569
  7. Casein kinase 2: an 'eminence grise' in cellular regulation?
    Biochim Biophys Acta. 1990 Sep 24;1054(3):267-84 PMID: 2207178
  8. The rel-associated pp40 protein prevents DNA binding of Rel and NF-kappa B: relationship with I kappa B beta and regulation by phosphorylation.
    Genes Dev. 1991 Aug;5(8):1464-76 PMID: 1907941
  9. Generation of p50 subunit of NF-kappa B by processing of p105 through an ATP-dependent pathway.
    Nature. 1991 Dec 5;354(6352):395-8 PMID: 1956402
  10. I kappa B gamma, a 70 kd protein identical to the C-terminal half of p110 NF-kappa B: a new member of the I kappa B family.
    Cell. 1992 Mar 20;68(6):1109-20 PMID: 1339305
  11. The ankyrin repeat domains of the NF-kappa B precursor p105 and the protooncogene bcl-3 act as specific inhibitors of NF-kappa B DNA binding.
    Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2489-93 PMID: 1532257
  12. Functional expression and growth factor activation of an epitope-tagged p44 mitogen-activated protein kinase, p44mapk.
    Mol Biol Cell. 1992 Jan;3(1):63-71 PMID: 1372523
  13. p105, the NF-kappa B p50 precursor protein, is one of the cellular proteins complexed with the v-Rel oncoprotein in transformed chicken spleen cells.
    J Virol. 1992 Jun;66(6):3758-67 PMID: 1533881
  14. Rapamycin-FKBP specifically blocks growth-dependent activation of and signaling by the 70 kd S6 protein kinases.
    Cell. 1992 Jun 26;69(7):1227-36 PMID: 1377606
  15. Candidate proto-oncogene bcl-3 encodes a subunit-specific inhibitor of transcription factor NF-kappa B.
    Nature. 1992 Aug 13;358(6387):597-9 PMID: 1501714
  16. I kappa B interacts with the nuclear localization sequences of the subunits of NF-kappa B: a mechanism for cytoplasmic retention.
    Genes Dev. 1992 Oct;6(10):1899-913 PMID: 1340770
  17. The candidate oncoprotein Bcl-3 is an antagonist of p50/NF-kappa B-mediated inhibition.
    Nature. 1992 Sep 24;359(6393):339-42 PMID: 1406939
  18. The precursor of NF-kappa B p50 has I kappa B-like functions.
    Cell. 1992 Oct 16;71(2):243-53 PMID: 1423592
  19. Mutational analysis of the p50 subunit of NF-kappa B and inhibition of NF-kappa B activity by trans-dominant p50 mutants.
    J Virol. 1993 Jan;67(1):288-93 PMID: 8416374
  20. The NF-kappa B precursor p105 and the proto-oncogene product Bcl-3 are I kappa B molecules and control nuclear translocation of NF-kappa B.
    EMBO J. 1993 Jan;12(1):213-22 PMID: 8428580
  21. p105 and p98 precursor proteins play an active role in NF-kappa B-mediated signal transduction.
    Genes Dev. 1993 Apr;7(4):705-18 PMID: 8458581
  22. Mutual regulation of the transcriptional activator NF-kappa B and its inhibitor, I kappa B-alpha.
    Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2532-6 PMID: 8460169
  23. Signal transduction via the MAP kinases: proceed at your own RSK.
    Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):5889-92 PMID: 8392180
  24. Common structural constituents confer I kappa B activity to NF-kappa B p105 and I kappa B/MAD-3.
    EMBO J. 1993 Jul;12(7):2781-8 PMID: 8334994
  25. Regulation of an epitope-tagged recombinant Rsk-1 S6 kinase by phorbol ester and erk/MAP kinase.
    Biochemistry. 1993 Aug 3;32(30):7727-38 PMID: 7688567
  26. Rapid proteolysis of I kappa B-alpha is necessary for activation of transcription factor NF-kappa B.
    Nature. 1993 Sep 9;365(6442):182-5 PMID: 8371761
  27. A growth factor-induced kinase phosphorylates the serum response factor at a site that regulates its DNA-binding activity.
    Mol Cell Biol. 1993 Oct;13(10):6260-73 PMID: 8413226
  28. In vivo control of NF-kappa B activation by I kappa B alpha.
    EMBO J. 1993 Dec;12(12):4685-95 PMID: 8223478
  29. Proteolytic degradation of MAD3 (I kappa B alpha) and enhanced processing of the NF-kappa B precursor p105 are obligatory steps in the activation of NF-kappa B.
    Nucleic Acids Res. 1993 Nov 11;21(22):5059-66 PMID: 8255759
  30. Distinct roles for cyclin-dependent kinases in cell cycle control.
    Science. 1993 Dec 24;262(5142):2050-4 PMID: 8266103
  31. Function and activation of NF-kappa B in the immune system.
    Annu Rev Immunol. 1994;12:141-79 PMID: 8011280
  32. Double-stranded RNA-dependent protein kinase activates transcription factor NF-kappa B by phosphorylating I kappa B.
    Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6288-92 PMID: 7912826
  33. A proteasome inhibitor prevents activation of NF-kappa B and stabilizes a newly phosphorylated form of I kappa B-alpha that is still bound to NF-kappa B.
    EMBO J. 1994 Nov 15;13(22):5433-41 PMID: 7957109
  34. Phosphorylation of I kappa B alpha precedes but is not sufficient for its dissociation from NF-kappa B.
    Mol Cell Biol. 1995 Mar;15(3):1302-11 PMID: 7862124
  35. NF-kappa B: a lesson in family values.
    Cell. 1995 Feb 24;80(4):529-32 PMID: 7867060
  36. Downregulation of MHC class I expression due to interference with p105-NF kappa B1 processing by Ad12E1A.
    EMBO J. 1995 Apr 3;14(7):1498-507 PMID: 7729425
  37. Inducible nuclear expression of newly synthesized I kappa B alpha negatively regulates DNA-binding and transcriptional activities of NF-kappa B.
    Mol Cell Biol. 1995 May;15(5):2689-96 PMID: 7739549
  38. Phosphorylation of human I kappa B-alpha on serines 32 and 36 controls I kappa B-alpha proteolysis and NF-kappa B activation in response to diverse stimuli.
    EMBO J. 1995 Jun 15;14(12):2876-83 PMID: 7796813
  39. RSK3 encodes a novel pp90rsk isoform with a unique N-terminal sequence: growth factor-stimulated kinase function and nuclear translocation.
    Mol Cell Biol. 1995 Aug;15(8):4353-63 PMID: 7623830
  40. Signal-induced site-specific phosphorylation targets I kappa B alpha to the ubiquitin-proteasome pathway.
    Genes Dev. 1995 Jul 1;9(13):1586-97 PMID: 7628694
  41. A role for phosphorylation and degradation in the control of NF-kappa B activity.
    Trends Genet. 1995 Jun;11(6):203-5 PMID: 7638895
  42. Constitutive phosphorylation of I kappa B alpha by casein kinase II.
    Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7637-41 PMID: 7644469
  43. Ubiquitin-mediated processing of NF-kappa B transcriptional activator precursor p105. Reconstitution of a cell-free system and identification of the ubiquitin-carrier protein, E2, and a novel ubiquitin-protein ligase, E3, involved in conjugation.
    J Biol Chem. 1995 Sep 15;270(37):21707-14 PMID: 7665588
  44. N- and C-terminal sequences control degradation of MAD3/I kappa B alpha in response to inducers of NF-kappa B activity.
    Mol Cell Biol. 1995 Oct;15(10):5339-45 PMID: 7565683
  45. Insulin activates nuclear factor kappa B in mammalian cells through a Raf-1-mediated pathway.
    J Biol Chem. 1995 Oct 13;270(41):24435-41 PMID: 7592658
  46. Evidence for two catalytically active kinase domains in pp90rsk.
    Mol Cell Biol. 1996 Mar;16(3):1212-9 PMID: 8622665
  47. Site-specific phosphorylation of IkappaBalpha by a novel ubiquitination-dependent protein kinase activity.
    Cell. 1996 Mar 22;84(6):853-62 PMID: 8601309
  48. Human T-cell lymphotropic virus type 1 Tax1 activation of NF-kappa B: involvement of the protein kinase C pathway.
    J Virol. 1996 Apr;70(4):2525-32 PMID: 8642682
  49. Mapping of the inducible IkappaB phosphorylation sites that signal its ubiquitination and degradation.
    Mol Cell Biol. 1996 Apr;16(4):1295-304 PMID: 8657102
  50. Phosphorylation of IkappaBalpha in the C-terminal PEST domain by casein kinase II affects intrinsic protein stability.
    Mol Cell Biol. 1996 Apr;16(4):1401-9 PMID: 8657113
  51. Characterization of IkappaB kinases. IkappaB-alpha is not phosphorylated by Raf-1 or protein kinase C isozymes, but is a casein kinase II substrate.
    J Biol Chem. 1996 Jun 7;271(23):13868-74 PMID: 8662925
  52. Dissection of TNF receptor 1 effector functions: JNK activation is not linked to apoptosis while NF-kappaB activation prevents cell death.
    Cell. 1996 Nov 1;87(3):565-76 PMID: 8898208
  53. Rel/NF-kappa B/I kappa B family: intimate tales of association and dissociation.
    Genes Dev. 1995 Nov 15;9(22):2723-35 PMID: 7590248
  54. The structure of an antigenic determinant in a protein.
    Cell. 1984 Jul;37(3):767-78 PMID: 6204768
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/16.11.3133
Published
1997-06-02
Pages
3133-44
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1169932
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/16.11.3133 PMC Full Text View journal details
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