Abstract
The immediate early transcription factor nuclear factor (IκBs) kappa B (NF-κB) is crucially involved in the regulation of numerous physiological or pathophysiological processes such as inflammation and tumourigenesis. Therefore, the control of NF-κB activity, which is mainly regulated by signal-induced degradation of cytoplasmic inhibitors of NF-κB (IκBs), is of high relevance. One known alternative pathway of NF-κB regulation is the stimulus-induced proteasomal degradation of RelB, a component of the NF-κB dimer. Here, we identified the serine/threonine protein kinase glycogen synthase kinase-3β (GSK-3β) as a critical signalling component leading to RelB degradation. In Jurkat leukaemic T cells as well as in primary human T cells, tetradecanoylphorbolacetate/ionomycin- and CD3/CD28-induced RelB degradation were impaired by a GSK-3β-specific pharmacological inhibitor, an ectopically expressed dominant-negative GSK-3β mutant and by small-interfering RNA-mediated silencing of GSK-3β expression. Furthermore, a physical interaction between RelB and GSK-3β was shown by co-immunoprecipitation, which was already notable in unstimulated cells. Most importantly, as demonstrated by in vitro kinase assays, human RelB is inducibly phosphorylated by GSK-3β, indicating a direct substrate–enzyme relationship. The serine residue 552 is a target of GSK-3β-mediated phosphorylation in vitro and in vivo. We conclude that GSK-3β is a crucial regulator of RelB degradation, stressing the relevant linkage between the NF-κB system and GSK-3β.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Avraham E, Szargel R, Eyal A, Rott R, Engelender S . (2005). Glycogen synthase kinase 3β modulates synphilin-1 ubiquitylation and cellular inclusion formation by SIAH: implications for proteasomal function and Lewy body formation. J Biol Chem 280: 42877–42886.
Bain J, Plater L, Elliott M, Shpiro N, Hastie CJ, McLauchlan H et al. (2007). The selectivity of protein kinase inhibitors: a further update. Biochem J 408: 297–315.
Barbie DA, Tamayo P, Boehm JS, Kim SY, Moody SE, Dunn IF et al. (2009). Systematic RNA interference reveals that oncogenic KRAS-driven cancers require TBK1. Nature 462: 108–112.
Beg AA, Sha WC, Bronson RT, Ghosh S, Baltimore D . (1995). Embryonic lethality and liver degeneration in mice lacking the RelA component of NF-κB. Nature 376: 167–170.
Billadeau D . (2007). Primers on molecular pathways: the glycogen synthase kinase-3β. Pancreatology 7: 398–402.
Buss H, Dörrie A, Schmitz ML, Frank R, Livingstone M, Resch K et al. (2004). Phosphorylation of serine 468 by GSK-3β negatively regulates basal p65 NF-κB activity. J Biol Chem 279: 49571–49574.
Compagno M, Lim WK, Grunn A, Nandula SV, Brahmachary M, Shen Q et al. (2009). Mutations of multiple genes cause deregulation of NF-κB in diffuse large B-cell lymphoma. Nature 459: 717–721.
Dar AA, Belkhiri A, El-Rifai W . (2009). The aurora kinase A regulates GSK-3β in gastric cancer cells. Oncogene 28: 866–875.
De Groot RP, Auwerx J, Bourouis M, Sassone-Corsi P . (1993). Negative regulation of Jun/AP-1: conserved function of glycogen synthase kinase 3 and the Drosophila kinase shaggy. Oncogene 8: 841–847.
Demarchi F, Bertoli C, Sandy P, Schneider C . (2003). GSK-3β regulates NF-κB1/p105 stability. J Biol Chem 278: 39583–39590.
Doble BW, Woodgett JR . (2003). GSK-3: tricks of the trade for a multi-tasking kinase. J Cell Sci 116: 1175–1186.
Fiol CJ, Mahrenholz AM, Wang Y, Roeske RW, Roach PJ . (1987). Formation of protein kinase recognition sites by covalent modification of the substrate. Molecular mechanism for the synergistic action of casein kinase II and glycogen synthase kinase-3. J Biol Chem 262: 14042–14048.
Forde JE, Dale TC . (2007). Glycogen synthase kinase 3: a key regulator of cellular fate. Cell Mol Life Sci 64: 1930–1944.
Hoeflich KP, Luo J, Rubie EA, Tsao MS, Jin O, Woodgett JR . (2000). Requirement for GSK-3β in cell survival and NF-κB activation. Nature 406: 86–90.
Karin M, Greten FR . (2005). NF-κB: linking inflammation and immunity to cancer development and progression. Nat Rev Immunol 5: 749–759.
Kato M, Sanada M, Kato I, Sato Y, Takita J, Takeuchi K et al. (2009). Frequent inactivation of A20 in B-cell lymphomas. Nature 459: 712–716.
Li Q, Van Antwerp D, Mercurio F, Lee KF, Verma IM . (1999). Severe liver degeneration in mice lacking the IκB kinase 2 gene. Science 284: 321–325.
Marienfeld R, Berberich-Siebelt F, Berberich I, Denk A, Serfling E, Neumann M . (2001). Signal-specific and phosphorylation-dependent RelB degradation: a potential mechanism of NF-κB control. Oncogene 20: 8142–8147.
Marienfeld R, May MJ, Berberich I, Serfling E, Ghosh S, Neumann M . (2003). RelB forms transcriptionally inactive complexes with RelA/p65. J Biol Chem 278: 19852–19860.
Meylan E, Dooley AL, Feldser DM, Shen L, Turk E, Ouyang C et al. (2009). Requirement for NF-κB signalling in a mouse model of lung adenocarcinoma. Nature 462: 104–107.
Neumann M, Naumann M . (2007). Beyond IκBs: alternative regulation of NF-κB activity. FASEB J 21: 2642–2654.
Ougolkov AV, Bone ND, Fernandez-Zapico ME, Kay NE, Billadeau DD . (2007). Inhibition of GSK-3 activity leads to epigenetic silencing of NF-κB target genes and induction of apoptosis in chronic lymphocytic leukemia B cells. Blood 110: 735–742.
Schwabe RF, Sakurai H . (2005). IKKβ phosphorylates p65 at S468 in transactivation domain 2. FASEB J 19: 1758–1760.
Sears R, Nuckolls F, Haura E, Taya Y, Tamai K, Nevins JR . (2000). Multiple Ras-dependent phosphorylation pathways regulate Myc protein stability. Genes Dev 14: 2501–2514.
Steinbrecher KA, Wilson III W, Cogswell PC, Baldwin AS . (2005). GSK-3β functions to specify gene-specific, NF-κB-dependent transcription. Mol Cell Biol 25: 8444–8455.
Viatour P, Dejardin E, Warnier M, Lair F, Claudio E, Bureau F et al. (2004). GSK3-mediated BCL-3 phosphorylation modulates its degradation and its oncogenicity. Mol Cell 16: 35–45.
Vincent T, Kukalev A, Andäng M, Pettersson R, Percipalle P . (2008). The glycogen synthase kinase (GSK) 3β represses RNA polymerase I transcription. Oncogene 27: 5254–5259.
Yuan J, Zhang J, Wong BW, Si X, Wong J, Yang D et al. (2005). Inhibition of glycogen synthase kinase 3β suppresses coxsackievirus-induced cytopathic effect and apoptosis via stabilization of β-catenin. Cell Death Differ 12: 1097–1106.
Acknowledgements
This work was supported by grants from the Deutsche Forschungsgemeinschaft (NE608/3-2 to MN; LE953/5-1 to ML) and by DFG grant SFB-TR19 (to KK and RK). We thank Sandra Bundschuh, Beatrix Kramer and Konstantin Klein for their excellent technical assistance.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Supplementary Information accompanies the paper on the Oncogene website
Rights and permissions
About this article
Cite this article
Neumann, M., Klar, S., Wilisch-Neumann, A. et al. Glycogen synthase kinase-3β is a crucial mediator of signal-induced RelB degradation. Oncogene 30, 2485–2492 (2011). https://doi.org/10.1038/onc.2010.580
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2010.580
Keywords
This article is cited by
-
GSK3β modulates NF-κB activation and RelB degradation through site-specific phosphorylation of BCL10
Scientific Reports (2018)
-
Decreased expression of the NF-κB family member RelB in lung fibroblasts from Smokers with and without COPD potentiates cigarette smoke-induced COX-2 expression
Respiratory Research (2015)
-
Transcriptional repression by the HDAC4–RelB–p52 complex regulates multiple myeloma survival and growth
Nature Communications (2015)