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Discovery of an NRF 1‐specific inducer from a large‐scale chemical library using a direct NRF 1‐protein monitoring system
Author(s) -
Tsujita Tadayuki,
Baird Liam,
Furusawa Yuki,
Katsuoka Fumiki,
Hou Yoshika,
Gotoh Satomi,
Kawaguchi Shinichi,
Yamamoto Masayuki
Publication year - 2015
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1111/gtc.12248
Subject(s) - nrf1 , biology , reporter gene , proteasome , biochemistry , transfection , microbiology and biotechnology , chemical library , luciferase , transcription factor , gene expression , gene , small molecule
NRF 1 ( NF ‐E2‐p45‐related factor 1) plays an important role in the regulation of genes encoding proteasome subunits, a cystine transporter, and lipid‐metabolizing enzymes. Global and tissue‐specific disruptions of the Nrf1 gene in mice result in embryonic lethality and spontaneous development of severe tissue defects, respectively, suggesting NRF 1 plays a critical role in vivo . Mechanistically, the continuous degradation of the NRF 1 protein by the proteasome is regarded as a major regulatory nexus of NRF 1 activity. To develop NRF 1‐specific inducers that act to overcome the phenotypes related to the lack of NRF 1 activity, we constructed a novel NRF 1ΔC‐Luc fusion protein reporter and developed cell lines that stably express the reporter in Hepa1c1c7 cells for use in high‐throughput screening. In screening of a chemical library with this reporter system, we identified two hit compounds that significantly induced luciferase activity. Through an examination of a series of derivatives of one of the hit compounds, we identified T1‐20, which induced a 70‐fold increase in luciferase activity. T1‐20 significantly increased the level of NRF 1 protein in the mouse liver, indicating that the compound is also functional in vivo . Thus, these results show the successful identification of the first small chemical compounds which specifically and significantly induce NRF 1.