Phosphorylation of Nrf2 at Ser-40 by Protein Kinase C Regulates Antioxidant Response Element-mediated Transcription*

By H.‐C. Huang, Truyen Nguyen, C. B. Pickett. Journal of Biological Chemistry, 2002

1,047 citations recorded by the source corpus.

Nrf2, a basic leucine zipper transcription factor, is an essential activator of the coordinated transcription of genes encoding antioxidant enzymes and phase II detoxifying enzymes through the regulatory sequence termed antioxidant response element (ARE). Recently we reported evidence for the involvement of protein kinase C (PKC) in phosphorylating Nrf2 and triggering its nuclear translocation in response to oxidative stress. We show here that phosphorylation of purified rat Nrf2 by the catalytic subunit of PKC was blocked by a synthetic peptide mimicking one of the potential PKC sites. Accordingly, Nrf2 bearing a Ser to Ala mutation at amino acid 40 (S40A) could not be phosphorylated by PKC. The S40A mutation did not affect in vitro binding of Nrf2/MafK to the ARE. However, it partially impaired Nrf2 activation of ARE-driven transcription in a reporter gene assay when Keap1 was overexpressed. In vitro transcribed/translated Keap1 could be coimmunoprecipitated with Nrf2. Phosphorylation of wild-type Nrf2 by PKC promoted its dissociation from Keap1, whereas the Nrf2-S40A mutant remained associated. These findings together with our prior studies suggest that the PKC-catalyzed phosphorylation of Nrf2 at Ser-40 is a critical signaling event leading to ARE-mediated cellular antioxidant response.

Subfields: Molecular Biology, Plant Science

View Paper: https://www.semanticscholar.org/paper/8966a9a65047c9e6ea380ed005ccb1ba6e2868b7

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Citations in the corpus

Redox Regulation of Cell Survival (2008, neutral): “Phosphorylation of Nrf2 promotes its dissociation from Keap1, allowing the free Nrf2 to translocate to nucleus (56, 126).”
Oxidative Stress and Oxidative Damage in Carcinogenesis (2010, background): “Secondly, activation of protein kinases, such as PKC, results in phosphorylation of Nrf2, which enhances the stability and/or release of Nrf2 from Keap1 (H. C. Huang, Nguyen, and Pickett 2002).”
Transcriptional Regulation by Nrf2 (2017, background): “Protein kinase C (PKC) phosphorylates Ser40 in the Neh2 domain of NRF2, disrupting KEAP1-NRF2 association and thus promoting NRF2 activation (55).”
Induction of phase I, II and III drug metabolism/transport by xenobiotics (2005, background): “Furthermore, Huang et aL have reported that protein kinase C (PKC) can directly phosphorylated Nrf2 (Huang et aL, 2000) and Ser-40 appears to be a site of potential phosphorylation (Huang et al., 2002).”
Nrf2 and Oxidative Stress: A General Overview of Mechanisms and Implications in Human Disease (2022, background): “Estrogen receptor-positive MCF7 breast cancer cells demonstrate overexpression of DPP3 which is associated with increased Nrf2 gene expression and poor prognosis [100].”
Novel mechanisms of natural antioxidant compounds in biological systems: involvement of glutathione and glutathione-related enzymes. (2005, background): “[143] Huang HC, Nguyen T, Pickett CB.”
Molecular mechanisms of the Keap1–Nrf2 pathway in stress response and cancer evolution (2011, background): “Ser40 and Tyr568 have been identified as phosphorylation sites of Nrf2 by PKC and tyrosine kinase Fyn, respectively (Huang et al. 2002; Bloom & Jaiswal 2003; Numazawa et al. 2003; Jain & Jaiswal 2006).”
The Molecular Mechanisms Regulating the KEAP1-NRF2 Pathway (2020, background): “of Ser40 phosphorylation by protein kinase C (PKC) (Table 4) (184, 185).”
NRF2, a Transcription Factor for Stress Response and Beyond (2020, background): “Extracellular signal-regulated protein kinases (ERK) [24], c-jun N-terminal kinase (JNK) [24], PI3K-AKT [19], protein kinase C (PKC) [25], casein kinase 2 (CK2) [26], and protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK) [27] were reported to mediate phosphorylation of NRF2 and increas”