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Updated: Dec 9, 2025

Identification of Transcription Factor Regulators using Medium-Throughput Screening of Arrayed Libraries and a Dual-Luciferase-Based Reporter
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A functionally defined high-density NRF2 interactome reveals new conditional regulators of ARE transactivation

Jonathan Poh1, Amy H Ponsford1, James Boyd1

  • 1Institute of Translational Medicine, University of Liverpool, UK.

Redox Biology
|September 10, 2020
PubMed

Insights

This study maps the human NRF2 interactome, identifying new partners that regulate its activity. Over 77% of these new partners modify NRF2 responses, offering insights into disease and drug resistance.

Area of Science:

  • Molecular Biology
  • Cellular Biology
  • Genetics

Background:

  • Nuclear factor erythroid 2-related factor 2 (NRF2) is a key transcription factor involved in cellular protection.
  • NRF2 dysregulation is implicated in over 60 human diseases, drug resistance, and adverse drug reactions.

Purpose of the Study:

  • To systematically map the human NRF2 interactome and identify novel NRF2-interacting partners.
  • To functionally characterize the impact of newly identified NRF2 partners on NRF2 activity in health and disease contexts.

Main Methods:

  • Systematic testing of 1962 predicted NRF2-partner interactions.
  • Verification using co-immunoprecipitation, dual luminescence-based co-immunoprecipitation (DULIP), and live-cell fluorescence cross-correlation spectroscopy (FCCS).
  • Functional assessment in NRF2 knockout (NRF2-/-) and disease-model (NRF2T80K, KEAP1-/-) cell lines.

Main Results:

  • An experimentally defined, high-density human NRF2 interactome was generated.
  • 46 new NRF2 partners were identified and verified.
  • Over 77% (17/22) of the newly investigated partners significantly modified NRF2 responses, with some effects specific to disease conditions.

Conclusions:

  • The study provides a comprehensive NRF2 interactome, revealing complex regulatory networks.
  • Newly identified NRF2 partners play crucial roles in modulating NRF2 activity, particularly under disease-related conditions.
  • This resource offers new perspectives on NRF2 function in health and disease, impacting therapeutic strategies.
Keywords:
Binary interactomeDual luminescence-based co-immunoprecipitation (DULIP)Fluorescence cross-correlation spectroscopy (FCCS)Human disease networkKEAP1NRF2/NFE2L2Protein interaction network (PIN)

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