Casein kinase II-mediated phosphorylation of NF-kappaB p65 subunit enhances inducible nitric-oxide synthase gene transcription in vivo

Aurélie Chantôme, Alena Pance, Nolwenn Gauthier, David Vandroux, Julie Chenu, Eric Solary, Jean-François Jeannin, Sylvie Reveneau

Research output: Contribution to journalArticlepeer-review


Nitric oxide (NO) produced by inducible nitric-oxide synthase (NOSII) is mainly regulated at the transcriptional level by the nuclear factor-kappaB (NF-kappaB). In the present study, we further analyzed the role of NF-kappaB in the in vivo transcriptional regulation of NOSII gene by comparing two clones isolated from the EMT-6 mouse mammary cancer cell line. In response to interleukin (IL)-1beta or lipopolysaccharide (LPS), EMT-6 clone J (EMT-6J) cells produce 3-fold more NO than EMT-6 clone H (EMT-6H) cells, an effect correlated with enhanced activation of NF-kappaB in EMT-6J cells. In response to IL-1beta, the kinetics of degradation of NF-kappaB inhibitors IkappaB-alpha and IkappaB-beta, the nucleo-cytoplasmic shuttling of the transcription factor and its binding to a specific DNA sequence were similar in both clones. In contrast, an IL-1beta-induced phosphorylation of serine residues in NF-kappaB p65 subunit was observed in EMT-6J, but not in EMT-6H, cells. This IL-1beta-induced phosphorylation of p65 was specifically prevented by pretreatment of EMT-6J cells with the casein kinase II inhibitor DRB. Small interfering RNA-mediated depletion of casein kinase II-alpha subunit also decreased NF-kappaB transcriptional activity and NOSII gene transcription in IL-1beta and LPS-stimulated EMT-6J cells to the levels observed in EMT-6H cells treated in the same conditions. Altogether, these data indicate that casein kinase II-mediated phosphorylation of p65 subunit can enhance the transcriptional activity of NF-kappaB in vivo. This post-translational modification of the transcription factor can be responsible for increased NOSII gene transcription and NO production in tumor cells exposed to either IL-1beta or LPS.

Original languageEnglish
Pages (from-to)23953-60
Number of pages8
JournalThe Journal of biological chemistry
Issue number23
Publication statusPublished - 4 Jun 2004


  • Animals
  • Blotting, Northern
  • Casein Kinase II
  • Cell Line
  • Cell Line, Tumor
  • Cell Nucleus/metabolism
  • Cytoplasm/metabolism
  • DNA/metabolism
  • Enzyme Activation
  • Genes, Reporter
  • Immunoblotting
  • Interleukin-1/metabolism
  • Kinetics
  • Lipopolysaccharides/metabolism
  • Mice
  • Microscopy, Fluorescence
  • Mutagenesis, Site-Directed
  • NF-kappa B/metabolism
  • Nitric Oxide/metabolism
  • Nitric Oxide Synthase/metabolism
  • Nitric Oxide Synthase Type II
  • Phosphorylation
  • Plasmids/metabolism
  • Protein Binding
  • Protein Processing, Post-Translational
  • Protein Serine-Threonine Kinases/metabolism
  • Protein Structure, Tertiary
  • RNA, Small Interfering/metabolism
  • Serine/chemistry
  • Time Factors
  • Transcription Factor RelA
  • Transcription, Genetic
  • Transcriptional Activation
  • Transfection


Dive into the research topics of 'Casein kinase II-mediated phosphorylation of NF-kappaB p65 subunit enhances inducible nitric-oxide synthase gene transcription in vivo'. Together they form a unique fingerprint.

Cite this