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Updated: Aug 10, 2026

Cell Aggregation Assays to Evaluate the Binding of the Drosophila Notch with Trans-Ligands and its Inhibition by Cis-Ligands
Published on: January 2, 2018
Notch(ic)-ER chimeras display hormone-dependent transformation, nuclear accumulation, phosphorylation and CBF1
1Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati, College of Medicine, Ohio 45267-0524, USA.
Notch proteins can act as oncogenes, driving cancer. This study shows inducible Notch intracellular domain (NICD)-estrogen receptor (ER) chimeras cause reversible cell transformation, requiring phosphorylation and nuclear localization for activity.
Area of Science:
- Molecular Biology
- Cell Biology
- Cancer Research
Background:
- Notch genes encode transmembrane receptors crucial for cellular processes like differentiation and proliferation.
- Notch signaling is vital in development and adult life.
- Three mammalian Notch genes function as oncogenes in tumors, but the transformation mechanism is unclear.
Purpose of the Study:
- To investigate early signaling events of Notch during cellular transformation.
- To construct and characterize inducible Notch intracellular domain (NICD)-estrogen receptor (ER) fusion proteins.
Main Methods:
- Created inducible NICD-ER alleles by fusing NICD to the ER hormone-binding domain.
- Utilized 4-Hydroxytamoxifen (OHT) for conditional activation of NICD-ER chimeras.
- Assessed transformation in clonal RKE cell lines expressing NICD-ER.
Main Results:
- NICD-ER chimeras were conditionally activated by OHT in a dose-dependent manner.
- NICD-ER expression induced hormone-dependent, reversible cell transformation in vitro.
- Activated NICD-ER chimeras were hyperphosphorylated and localized to the nucleus.
Conclusions:
- Notch proteins' oncogenic activity can be studied using inducible NICD-ER systems.
- Cellular transformation by Notch requires protein hyperphosphorylation and nuclear accumulation.
- Chronic stimulation is necessary for maintaining the transformed phenotype induced by Notch.
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