Interaction of PKN with a neuron-specific basic helix-loop-helix transcription factor, NDRF/NeuroD2.

By the yeast two-hybrid screening of a human brain cDNA library with the amino-terminal regulatory region of PKN as a bait, a clone encoding a neuron-specific basic Helix-Loop-Helix (bHLH) transcription factor, NDRF/NeuroD2 was isolated. NDRF/NeuroD2 was co-precipitated with PKN from the lysate of COS-7 cells transfected with both expression constructs ...
for NDRF/NeuroD2 and PKN. In vitro binding studies using the deletion mutants of NDRF/NeuroD2 synthesized in a rabbit reticulocyte lysate indicated that the internal region containing the bHLH domain of NDRF/NeuroD2 was necessary and sufficient for the interaction with PKN. In addition, recombinant NDRF/NeuroD2 purified from Escherichia coli could bind PKN, suggesting the direct interaction between NDRF/NeuroD2 and PKN. Transient transfection assays using P19 cells revealed that expression of NDRF/NeuroD2 increased the transactivation of the rat insulin promoter element 3 (RIPE3) enhancer up to approximately 12-fold and that co-expression of catalytically active form of PKN, but not kinase-deficient derivative, resulted in a further threefold increase of NDRF/NeuroD2-mediated transcription. These findings suggest that PKN may contribute to transcriptional responses through the post-translational modification of the NDRF/NeuroD2-dependent transcriptional machinery.
Mesh Terms:
Animals, Basic Helix-Loop-Helix Transcription Factors, COS Cells, Catalysis, Cell Line, Gene Expression Regulation, Helix-Loop-Helix Motifs, Humans, Mutation, Neuropeptides, Protein Binding, Protein Kinase C, Protein-Serine-Threonine Kinases, Protein-Tyrosine Kinases, Rabbits, Rats, Recombinant Fusion Proteins, Transcription Factors, Transcription, Genetic, Tumor Cells, Cultured, Two-Hybrid System Techniques
Brain Res. Mol. Brain Res.
Date: Dec. 10, 1999
Download Curated Data For This Publication
10055
Switch View:
  • Interactions 3