Insulin Activates Nuclear Factor κB in Mammalian Cells through a Raf-1-mediated Pathway

Autor: Gisèle Cherqui, P J Antoine, Carole Philippe, Laurent Baud, Jacqueline Capeau, André Groyer, F. Bertrand
Rok vydání: 1995
Předmět:
Pyrrolidines
Sodium Salicylate
Molecular Sequence Data
8-Bromo Cyclic Adenosine Monophosphate
CHO Cells
Protein Serine-Threonine Kinases
Transfection
Biochemistry
Antioxidants
Thiocarbamates
Cricetinae
Proto-Oncogene Proteins
Insulin receptor substrate
Animals
Humans
Insulin
Point Mutation
Cycloheximide
Enzyme Inhibitors
Insulin-Like Growth Factor I
Phosphorylation
Kinase activity
Molecular Biology
Protein kinase B
Binding Sites
Aspirin
Base Sequence
biology
GRB10
Transcription Factor RelB
Autophosphorylation
NF-kappa B
Cell Biology
Molecular biology
Receptor
Insulin
Recombinant Proteins
IRS2
Proto-Oncogene Proteins c-raf
Kinetics
Insulin receptor
Mutagenesis
Site-Directed
biology.protein
Tetradecanoylphorbol Acetate
Tyrosine
Oligonucleotide Probes
Tyrosine kinase
Protein Binding
Signal Transduction
Transcription Factors
Zdroj: Journal of Biological Chemistry. 270:24435-24441
ISSN: 0021-9258
DOI: 10.1074/jbc.270.41.24435
Popis: We examined the effect of insulin on nuclear factor kappa B (NF-kappa B) activity in Chinese ovary (CHO) cells overexpressing wild-type (CHO-R cells) or -defective insulin receptors mutated at Tyr1162 and Tyr1163 autophosphorylation sites (CHO-Y2 cells). In CHO-R cells, insulin caused a specific, time-, and concentration-dependent activation of NF-kappa B. The insulin-induced DNA-binding complex was identified as the p50/p65 heterodimer. Insulin activation of NF-kappa B: 1) was related to insulin receptor number and tyrosine kinase activity since it was markedly reduced in parental CHO cells which proved to respond to insulin growth factor-1 and phorbol 12-myristate 13-acetate (PMA) activation, and was dramatically decreased in CHO-Y2 cells; 2) persisted in the presence of cycloheximide and was blocked by pyrrolidine dithiocarbamate, aspirin and sodium salicylate, three compounds interfering with I kappa B degradation and/or NF-kappa B.I kappa B complex dissociation; 3) was independent of both PMA-sensitive and atypical (zeta) protein kinases C; and 4) was dependent on Raf-1 kinase activity since insulin-stimulated NF-kappa B DNA binding activity was inhibited by 8-bromo-cAMP, a Raf-1 kinase inhibitor. Moreover, insulin activation of NF-kappa B-driven luciferase reporter gene expression was blocked in CHO-R cells expressing a Raf-1 dominant negative mutant. This is the first evidence that insulin activates NF-kappa B in mammalian cells through a post-translational mechanism requiring both insulin receptor tyrosine kinase and Raf-1 kinase activities.
Databáze: OpenAIRE
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