TAK1-TAB1 fusion protein: a novel constitutively active mitogen-activated protein kinase kinase kinase that stimulates AP-1 and NF-kappaB signaling pathways

Biochem Biophys Res Commun. 2002 Oct 11;297(5):1277-81. doi: 10.1016/s0006-291x(02)02379-3.

Abstract

TAK1 mitogen-activated protein kinase kinase kinase (MAP3K) is activated by its specific activator, TAK1-binding protein 1 (TAB1). A constitutively active TAK1 mutant has not yet been generated due to the indispensable requirement of TAB1 for TAK1 kinase activity. In this study, we generated a novel constitutively active TAK1 by fusing its kinase domain to the minimal TAK1-activation domain of TAB1. Co-immunoprecipitation assay demonstrated that these domains interacted intra-molecularly. The TAK1-TAB1 fusion protein showed a significant MAP3K activity in vitro and activated c-Jun N-terminal kinase/p38 MAPKs and IkappaB kinase in vivo, which was followed by increased production of interleukin-6. These results indicate that the fusion protein is useful for characterizing the physiological roles of the TAK1-TAB1 complex.

MeSH terms

  • Adaptor Proteins, Signal Transducing*
  • Binding Sites
  • Blotting, Western
  • Carrier Proteins / chemistry*
  • Carrier Proteins / metabolism
  • DNA, Complementary / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • HeLa Cells
  • Humans
  • Interleukin-6 / metabolism
  • Interleukin-8 / metabolism
  • Intracellular Signaling Peptides and Proteins*
  • JNK Mitogen-Activated Protein Kinases
  • MAP Kinase Kinase Kinases / chemistry*
  • MAP Kinase Kinase Kinases / metabolism
  • MAP Kinase Signaling System
  • Mitogen-Activated Protein Kinases / metabolism
  • Models, Genetic
  • Mutation
  • NF-kappa B / metabolism*
  • Precipitin Tests
  • Protein Binding
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / metabolism
  • Transcription Factor AP-1 / metabolism*
  • Transfection
  • p38 Mitogen-Activated Protein Kinases

Substances

  • Adaptor Proteins, Signal Transducing
  • Carrier Proteins
  • DNA, Complementary
  • Interleukin-6
  • Interleukin-8
  • Intracellular Signaling Peptides and Proteins
  • NF-kappa B
  • Recombinant Fusion Proteins
  • TAB1 protein, MAPKKK activator, vertebrate
  • TAB1 protein, human
  • Transcription Factor AP-1
  • JNK Mitogen-Activated Protein Kinases
  • Mitogen-Activated Protein Kinases
  • p38 Mitogen-Activated Protein Kinases
  • MAP Kinase Kinase Kinases
  • MAP kinase kinase kinase 7