Related Experiment Video

Updated: Aug 10, 2026

Cell Aggregation Assays to Evaluate the Binding of the Drosophila Notch with Trans-Ligands and its Inhibition by Cis-Ligands
05:48

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

C Ronchini1, A J Capobianco

  • 1Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati, College of Medicine, Ohio 45267-0524, USA.

Oncogene
|August 22, 2000
PubMed

Insights

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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