A functionally defined high-density NRF2 interactome reveals new conditional regulators of ARE transactivation



Poh, Jonathan, Ponsford, Amy H ORCID: 0000-0002-7178-7862, Boyd, James ORCID: 0000-0003-0858-7179, Woodsmith, Jonathan, Stelzl, Ulrich, Wanker, Erich, Harper, Nicholas, MacEwan, David ORCID: 0000-0002-2879-0935 and Sanderson, Christopher M ORCID: 0000-0002-7301-3537
(2020) A functionally defined high-density NRF2 interactome reveals new conditional regulators of ARE transactivation. Redox Biology, 37.

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Abstract

NRF2 (NFE2L2) is a cytoprotective transcription factor associated with >60 human diseases, adverse drug reactions and therapeutic resistance. To provide insight into the complex regulation of NRF2 responses, 1962 predicted NRF2-partner interactions were systematically tested to generate an experimentally defined high-density human NRF2 interactome. Verification and conditional stratification of 46 new NRF2 partners was achieved by co-immunoprecipitation and the novel integration of quantitative data from dual luminescence-based co-immunoprecipitation (DULIP) assays and live-cell fluorescence cross-correlation spectroscopy (FCCS). The functional impact of new partners was then assessed in genetically edited loss-of-function (NRF2−/−) and disease-related gain-of-function (NRF2T80K and KEAP1−/−) cell-lines. Of the new partners investigated >77% (17/22) modified NRF2 responses, including partners that only exhibited effects under disease-related conditions. This experimentally defined binary NRF2 interactome provides a new vision of the complex molecular networks that govern the modulation and consequence of NRF2 activity in health and disease.

Item Type: Article
Uncontrolled Keywords: NRF2/NFE2L2, KEAP1, Protein interaction network (PIN), Fluorescence cross-correlation spectroscopy (FCCS), Dual luminescence-based co-immunoprecipitation (DULIP), Human disease network, Binary interactome
Depositing User: Symplectic Admin
Date Deposited: 22 Sep 2020 08:44
Last Modified: 21 Jan 2021 12:14
DOI: 10.1016/j.redox.2020.101686
Open Access URL: https://doi.org/10.1016/j.redox.2020.101686
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3102150