I-kappa B Kinase; NF-kappa B; NF-KappaB Inhibitor alpha; Humans; Phosphorylation; Protein Binding; Signal Transduction; NF-KappaB Inhibitor alpha/metabolism; NF-KappaB Inhibitor alpha/genetics; Molecular Docking Simulation; HEK293 Cells; Substrate Specificity; I-kappa B Kinase/metabolism; I-kappa B Kinase/chemistry; I-kappa B Kinase/genetics; NF-kappa B/metabolism; Protein Multimerization; Amino Acid Motifs
Abstract :
[en] The inhibitor of κB (IκB) kinase (IKK) is a central regulator of NF-κB signaling. All IKK complexes contain hetero- or homodimers of the catalytic IKKβ and/or IKKα subunits. Here, we identify a YDDΦxΦ motif, which is conserved in substrates of canonical (IκBα, IκBβ) and alternative (p100) NF-κB pathways, and which mediates docking to catalytic IKK dimers. We demonstrate a quantitative correlation between docking affinity and IKK activity related to IκBα phosphorylation/degradation. Furthermore, we show that phosphorylation of the motif's conserved tyrosine, an event previously reported to promote IκBα accumulation and inhibition of NF-κB gene expression, suppresses the docking interaction. Results from integrated structural analyzes indicate that the motif binds to a groove at the IKK dimer interface. Consistently, suppression of IKK dimerization also abolishes IκBα substrate binding. Finally, we show that an optimized bivalent motif peptide inhibits NF-κB signaling. This work unveils a function for IKKα/β dimerization in substrate motif recognition.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Li, Changqing; Biotechnology and Cell Signaling (CNRS/Université de Strasbourg, UMR7242), Ecole Superieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, 67400, Illkirch, France
Moro, Stefano; Biotechnology and Cell Signaling (CNRS/Université de Strasbourg, UMR7242), Ecole Superieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, 67400, Illkirch, France
Shostak, Kateryna ; Université de Liège - ULiège > Département de pharmacie > Chimie médicale
O'Reilly, Francis J ; Institute of Biotechnology, Technische Universität Berlin, Gustav-Meyer-Allee 25, Berlin, Germany
Donzeau, Mariel; Biotechnology and Cell Signaling (CNRS/Université de Strasbourg, UMR7242), Ecole Superieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, 67400, Illkirch, France
Graziadei, Andrea; Institute of Biotechnology, Technische Universität Berlin, Gustav-Meyer-Allee 25, Berlin, Germany
McEwen, Alastair G ; Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC) / INSERM UMR-S 1258 / CNRS UMR7104/ Université de Strasbourg, 1 rue Laurent Fries, 67400, Illkirch, France
Desplancq, Dominique; Biotechnology and Cell Signaling (CNRS/Université de Strasbourg, UMR7242), Ecole Superieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, 67400, Illkirch, France
Poussin-Courmontagne, Pierre ; Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC) / INSERM UMR-S 1258 / CNRS UMR7104/ Université de Strasbourg, 1 rue Laurent Fries, 67400, Illkirch, France
Bachelart, Thomas; Biotechnology and Cell Signaling (CNRS/Université de Strasbourg, UMR7242), Ecole Superieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, 67400, Illkirch, France
Fiskin, Mert; Biotechnology and Cell Signaling (CNRS/Université de Strasbourg, UMR7242), Ecole Superieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, 67400, Illkirch, France
Berrodier, Nicolas; Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC) / INSERM UMR-S 1258 / CNRS UMR7104/ Université de Strasbourg, 1 rue Laurent Fries, 67400, Illkirch, France
Pichard, Simon ; Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC) / INSERM UMR-S 1258 / CNRS UMR7104/ Université de Strasbourg, 1 rue Laurent Fries, 67400, Illkirch, France
Brillet, Karl ; Institut Biologie Moléculaire et Cellulaire (IBMC), CNRS UPR9002, 2 allée Konrad Roentgen, 67000, Strasbourg, France
Orfanoudakis, Georges; Biotechnology and Cell Signaling (CNRS/Université de Strasbourg, UMR7242), Ecole Superieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, 67400, Illkirch, France
Poterszman, Arnaud ; Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC) / INSERM UMR-S 1258 / CNRS UMR7104/ Université de Strasbourg, 1 rue Laurent Fries, 67400, Illkirch, France
Torbeev, Vladimir; Biotechnology and Cell Signaling (CNRS/Université de Strasbourg, UMR7242), Ecole Superieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, 67400, Illkirch, France
Rappsilber, Juri ; Institute of Biotechnology, Technische Universität Berlin, Gustav-Meyer-Allee 25, Berlin, Germany
Davey, Norman E; Division of Cancer Biology, The Institute of Cancer Research, 237 Fulham Road, London, SW3 6JB, UK
Chariot, Alain ; Université de Liège - ULiège > Département de pharmacie > Chimie médicale ; WELBIO department, WEL Research Institute, avenue Pasteur, 6, 1300, Wavre, Belgium
Zanier, Katia ; Biotechnology and Cell Signaling (CNRS/Université de Strasbourg, UMR7242), Ecole Superieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, 67400, Illkirch, France. zanier@unistra.fr
ANR - Agence Nationale de la Recherche [FR] FRM - Fondation pour la Recherche Médicale [FR]
Funding text :
The work
was supported by grants from Agence Nationale de la Recherche (ANR
JCJC, ref. ANR-13-JSV8-0004-01), ‘Ligue contre le Cancer’ (CCIRGE),
‘Fondation pour La Recherche Medicale’ (Equipes FRM, ref.
DEQ20180339231), ITI Innovec (IdEx ANR-10-IDEX-0002), STRAT’US(
ANR-20-SFRI-0012)) and ‘Alsace contre le Cancer’ and Instruct-ERIC
(PID 11382) to KZ. Additional funding sources include ITI IMCBio+
(IdEx ANR-10-IDEX-0002, SFRI-STRAT’US ANR-20-SFRI-0012 and EUR
IMCBio, ANR-17-EURE-0023) and FRISBI (ANR-10-INBS-0005) to
IGBMC-CBI.
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