Tumor necrosis factor-alpha-induced IKK phosphorylation of NF-kappaB p65 on serine 536 is mediated through the TRAF2, TRAF5, and TAK1 signaling pathway - PubMed (original) (raw)

. 2003 Sep 19;278(38):36916-23.

doi: 10.1074/jbc.M301598200. Epub 2003 Jul 3.

Affiliations

• PMID: 12842894
• DOI: 10.1074/jbc.M301598200

Free article

Tumor necrosis factor-alpha-induced IKK phosphorylation of NF-kappaB p65 on serine 536 is mediated through the TRAF2, TRAF5, and TAK1 signaling pathway

Hiroaki Sakurai et al. J Biol Chem. 2003.

Free article

Abstract

The activation of NF-kappaB has been shown to be regulated by multiple phosphorylations of IkappaBs and the NF-kappaB p65 subunit. Here, we characterized the intracellular signaling pathway leading to phosphorylation of p65 on Ser-536 using a novel anti-phospho-p65 (Ser-536) antibody. The Ser-536 of endogenous p65 was rapidly phosphorylated in response to a wide variety of NF-kappaB stimulants including TNF-alpha in the cytoplasm and rapidly dephosphorylated in the nucleus. The TNF-alpha-but not IL-1beta-induced Ser-536 phosphorylation was severely impaired in murine embryonic fibroblasts derived from traf2-/-traf5-/- mice. Bay 11-7082, an inhibitor of IkappaB phosphorylation, inhibited the TNF-alpha-induced phosphorylation in vivo. In addition, overexpression of TGF-beta-activated kinase 1 (TAK1), IKKalpha and IKKbeta stimulated the phosphorylation, and their dominant negative mutants blocked the TNF-alpha-induced phosphorylation. Moreover, small interfering RNAs (siRNAs) against TAK1, IKKalpha and IKKbeta blocked the phosphorylation of endogenous p65. On the other hand, calyculin-A, a protein phosphatase inhibitor, blocked the dephosphorylation in the nucleus in vivo. These results indicate that similar signaling pathways were utilized for the phosphorylations of IkappaBalpha and p65, which further support the idea that both IkappaB and NF-kappaB are substrates for the IKK complex in the activation of NF-kappaB.

PubMed Disclaimer

Similar articles

• Constitutive and interleukin-1-inducible phosphorylation of p65 NF-{kappa}B at serine 536 is mediated by multiple protein kinases including I{kappa}B kinase (IKK)-{alpha}, IKK{beta}, IKK{epsilon}, TRAF family member-associated (TANK)-binding kinase 1 (TBK1), and an unknown kinase and couples p65 to TATA-binding protein-associated factor II31-mediated interleukin-8 transcription.
  Buss H, Dörrie A, Schmitz ML, Hoffmann E, Resch K, Kracht M. Buss H, et al. J Biol Chem. 2004 Dec 31;279(53):55633-43. doi: 10.1074/jbc.M409825200. Epub 2004 Oct 15. J Biol Chem. 2004. PMID: 15489227
• IkappaB kinases phosphorylate NF-kappaB p65 subunit on serine 536 in the transactivation domain.
  Sakurai H, Chiba H, Miyoshi H, Sugita T, Toriumi W. Sakurai H, et al. J Biol Chem. 1999 Oct 22;274(43):30353-6. doi: 10.1074/jbc.274.43.30353. J Biol Chem. 1999. PMID: 10521409
• TAK1 is critical for IkappaB kinase-mediated activation of the NF-kappaB pathway.
  Takaesu G, Surabhi RM, Park KJ, Ninomiya-Tsuji J, Matsumoto K, Gaynor RB. Takaesu G, et al. J Mol Biol. 2003 Feb 7;326(1):105-15. doi: 10.1016/s0022-2836(02)01404-3. J Mol Biol. 2003. PMID: 12547194
• Regulation and function of IKK and IKK-related kinases.
  Häcker H, Karin M. Häcker H, et al. Sci STKE. 2006 Oct 17;2006(357):re13. doi: 10.1126/stke.3572006re13. Sci STKE. 2006. PMID: 17047224 Review.
• Beyond NF-κB activation: nuclear functions of IκB kinase α.
  Huang WC, Hung MC. Huang WC, et al. J Biomed Sci. 2013 Jan 23;20(1):3. doi: 10.1186/1423-0127-20-3. J Biomed Sci. 2013. PMID: 23343355 Free PMC article. Review.

Cited by

• Sex-specific differences in neonatal hyperoxic lung injury.
  Lingappan K, Jiang W, Wang L, Moorthy B. Lingappan K, et al. Am J Physiol Lung Cell Mol Physiol. 2016 Aug 1;311(2):L481-93. doi: 10.1152/ajplung.00047.2016. Epub 2016 Jun 24. Am J Physiol Lung Cell Mol Physiol. 2016. PMID: 27343189 Free PMC article.
• Oxidative Stress and Its Consequences in the Blood of Rats Irradiated with UV: Protective Effect of Cannabidiol.
  Biernacki M, Brzóska MM, Markowska A, Gałażyn-Sidorczuk M, Cylwik B, Gęgotek A, Skrzydlewska E. Biernacki M, et al. Antioxidants (Basel). 2021 May 21;10(6):821. doi: 10.3390/antiox10060821. Antioxidants (Basel). 2021. PMID: 34063802 Free PMC article.
• Nod1 acts as an intracellular receptor to stimulate chemokine production and neutrophil recruitment in vivo.
  Masumoto J, Yang K, Varambally S, Hasegawa M, Tomlins SA, Qiu S, Fujimoto Y, Kawasaki A, Foster SJ, Horie Y, Mak TW, Núñez G, Chinnaiyan AM, Fukase K, Inohara N. Masumoto J, et al. J Exp Med. 2006 Jan 23;203(1):203-13. doi: 10.1084/jem.20051229. Epub 2006 Jan 17. J Exp Med. 2006. PMID: 16418393 Free PMC article.
• A Three Stage Integrative Pathway Search (TIPS) framework to identify toxicity relevant genes and pathways.
  Li Z, Srivastava S, Mittal S, Yang X, Sheng L, Chan C. Li Z, et al. BMC Bioinformatics. 2007 Jun 14;8:202. doi: 10.1186/1471-2105-8-202. BMC Bioinformatics. 2007. PMID: 17570844 Free PMC article.
• The Roles of TNF Signaling Pathways in Metabolism of Bone Tumors.
  Zhou H, Dong Y, Alhaskawi A, Lai J, Wang Z, Ezzi SHA, Kota VG, Abdulla MHAH, Sun Z, Lu H. Zhou H, et al. Front Pharmacol. 2022 Jun 29;13:907629. doi: 10.3389/fphar.2022.907629. eCollection 2022. Front Pharmacol. 2022. PMID: 35847045 Free PMC article.

Publication types

MeSH terms

Substances

LinkOut - more resources

• Full Text Sources

  • Elsevier Science

• Other Literature Sources

  • The Lens - Patent Citations Database

• Molecular Biology Databases

  • BioCyc
  • Gene Ontology
  • GlyGen glycoinformatics resource
  • IMEx Consortium
  • PhosphoSitePlus

• Miscellaneous

  • NCI CPTAC Assay Portal