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

Autor: Amy H. Ponsford, James Boyd, Ulrich Stelzl, David J. MacEwan, Nicholas Harper, Christopher M. Sanderson, Erich E. Wanker, Jonathan Woodsmith, Jonathan Poh
Jazyk: angličtina
Rok vydání: 2020
Předmět:
0301 basic medicine
Transcriptional Activation
NF-E2-Related Factor 2
Clinical Biochemistry
High density
Functional impact
Computational biology
Biology
Protein interaction network (PIN)
Biochemistry
Interactome
digestive system
environment and public health
Fluorescence cross-correlation spectroscopy (FCCS)
Cell Line
03 medical and health sciences
Transactivation
0302 clinical medicine
Humans
Drug reaction
Transcription factor
lcsh:QH301-705.5
lcsh:R5-920
Kelch-Like ECH-Associated Protein 1
Organic Chemistry
Dual luminescence-based co-immunoprecipitation (DULIP)
respiratory system
NFE2L2
KEAP1
Molecular network
030104 developmental biology
Gene Expression Regulation
lcsh:Biology (General)
NRF2/NFE2L2
Human disease network
Function and Dysfunction of the Nervous System
lcsh:Medicine (General)
030217 neurology & neurosurgery
Binary interactome
Research Paper
Zdroj: Redox Biology, Vol 37, Iss, Pp 101686-(2020)
Redox Biology
ISSN: 2213-2317
Popis: 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.
Graphical abstract Image 1
Highlights • 1,962 predicted NRF2 partners were tested and 46 new binary partners were identified. • 77% of tested new NRF2 partners modulate ARE-mediated gene expression. • Some novel partners only exhibit functional effects under disease-like conditions. • Quantitative evidence that KEAP1 can modulate ARE-transactivation independently of NRF2. • New NRF2 binding partners reveal 130 novel disease gene associations.
Databáze: OpenAIRE
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