WDR23 regulates the expression of Nrf2-driven drug-metabolizing enzymes

Ferbian Milas Siswanto; Ami Oguro; Saki Arase; Susumu Imaoka

doi:10.1016/j.dmpk.2020.06.007

WDR23 regulates the expression of Nrf2-driven drug-metabolizing enzymes

Drug Metab Pharmacokinet. 2020 Oct;35(5):441-455. doi: 10.1016/j.dmpk.2020.06.007. Epub 2020 Jun 21.

Authors

Ferbian Milas Siswanto¹, Ami Oguro², Saki Arase¹, Susumu Imaoka³

Affiliations

¹ Department of Biomedical Chemistry, School of Science and Technology, Kwansei Gakuin University, Gakuen 2-1, Sanda, 669-1337, Japan.
² Department of Biomedical Chemistry, School of Science and Technology, Kwansei Gakuin University, Gakuen 2-1, Sanda, 669-1337, Japan; Program of Biomedical Science, Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, 739-8521, Japan.
³ Department of Biomedical Chemistry, School of Science and Technology, Kwansei Gakuin University, Gakuen 2-1, Sanda, 669-1337, Japan. Electronic address: imaoka@kwansei.ac.jp.

PMID: 32839090
DOI: 10.1016/j.dmpk.2020.06.007

Abstract

Nrf2 plays a central role in the response to xenobiotics and oxidative stress. The activation of Nrf2 induces the expression of drug-metabolizing enzymes (DMEs) and is important for cytoprotection. Keap1 is a widely accepted proteasome-dependent regulator of Nrf2. Keap1 was reported to be absent in Caenorhabditis elegans, and the level of the Nrf2 ortholog SKN-1 was mainly regulated by WDR23. The WDR23 locus is highly conserved from C. elegans to humans. We investigated whether WDR23 regulates Nrf2 activity in mammalian cells, hepatocellular carcinoma cells (Hep3B) and human cervical carcinoma cells (HeLa). We found that WDR23 has two isoforms (1 and 2) and that knockdown of WDR23 was sufficient to stabilize Nrf2 and alter the expression of several DMEs. Keap1 knockdown resulted in higher Nrf2 levels than WDR23 knockdown, and their effects on DMEs differed. These results were consistent with Keap1 being a canonical regulator of Nrf2, and that WDR23 may assist in Nrf2 regulation. We confirmed that WDR23 physically interacted with Nrf2, suggesting that WDR23 directly regulates Nrf2-dependent DMEs. In immunostaining experiments, human WDR23 isoform 1 was localized to the cytoplasm, whereas isoform 2 mainly resided in the nucleus. Taken together, our results suggested WDR23 is a novel regulator of DME expression.

Keywords: Drug-metabolizing enzymes; Keap1; Nrf2; WDR23.

MeSH terms

HeLa Cells
Humans
Kelch-Like ECH-Associated Protein 1 / metabolism*
NF-E2-Related Factor 2 / genetics
NF-E2-Related Factor 2 / metabolism*
Tumor Cells, Cultured
Ubiquitin-Protein Ligase Complexes / metabolism*

Substances

KEAP1 protein, human
Kelch-Like ECH-Associated Protein 1
NF-E2-Related Factor 2
NFE2L2 protein, human
DDB1 and CUL4 associated factor 11, human
Ubiquitin-Protein Ligase Complexes