RRC ID 27204
Author Kurooka H, Kuroda K, Honjo T.
Title Roles of the ankyrin repeats and C-terminal region of the mouse notch1 intracellular region.
Journal Nucleic Acids Res
Abstract The Notch intracellular region (RAMIC) interacts with a DNA binding protein RBP-J to activate transcription of genes that inhibit cell differentiation. The RAM domain and ankyrin (ANK) repeats of mouse Notch1 RAMIC were shown to be responsible for RBP-J binding and necessary for transactivation. The C-terminal portion of Notch1 RAMIC has also been suggested to be important for transactivation. Using GAL4 fusion constructs, we identified a novel transactivation domain (TAD) between the ANK repeats and the PEST sequence of mouse Notch1. The C-terminal half of mouse Notch2 RAMIC also exhibited TAD activity. Unexpectedly, the RBP-J chimeric protein with the Notch1 TAD failed to activate transcription but the activity was recovered by addition of either the RAM domain or ANK repeats. The results suggest that the activity of Notch1 TAD is repressed by fusion with RBP-J because of the presence of a RBP-J-associated co-repressor(s), which could be displaced by either the RAM domain or ANK repeats. Taken together, mouse Notch1 RAMIC can experimentally be separated into three functional domains: the RAM domain and ANK repeats for RBP-J binding and co-repressor displacement and the C-terminal TAD.
Volume 26(23)
Pages 5448-55
Published 1998-12-1
DOI 10.1093/nar/26.23.5448
PII gkb849
Description Original paper.
PMID 9826771
PMC PMC147984
MeSH 3T3 Cells Animals Ankyrins / physiology* COS Cells Cells, Cultured DNA-Binding Proteins / genetics DNA-Binding Proteins / physiology Immunoglobulin J Recombination Signal Sequence-Binding Protein Intracellular Fluid / chemistry Intracellular Fluid / physiology* Membrane Proteins / antagonists & inhibitors Membrane Proteins / genetics Membrane Proteins / physiology* Mice Nuclear Proteins* Receptor, Notch1 Receptors, Cell Surface* Recombinant Fusion Proteins / pharmacology Repetitive Sequences, Amino Acid / physiology* Repressor Proteins / pharmacology Sequence Deletion Transcription Factors* Transcriptional Activation
IF 11.147
Times Cited 156
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
DNA material pGa981-6 (RDB06776)