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Regulation of Nrf2 Transactivation Domain Activity
Open accessAbstract
Transcription factor NF-E2-related factor 2 (Nrf2) regulates the induction of Phase II detoxifying enzymes as well as anti-oxidative enzymes. In this study, we investigated the transactivation potential of different Nrf2 transactivation domain regions by using the Gal4-Nrf2 chimeras and Gal4-Luc reporter co-transfection assay system in HepG2 cells. The results indicated that chimera Gal4-Nrf2-(1-370), which contains the full transactivation domain showed very potent transactivation activity. The high transactivation activity of Gal4-Nrf2-(113-251) and the diminished transactivation activities of chimera Gal4-Nrf2-(1-126) and Gal4-Nrf2-(230-370) suggested that the Nrf2 N-terminal 113-251 amino acids region is critical in maintaining its transactivation activity. Overexpression of upstream MAPKs such as Raf, MEKK1, TAK1-DeltaN, and ASK1 up-regulated the transactivation activities of Gal4-Nrf2-(1-370) and Gal4-Nrf2-(113-251) in a dose-dependent manner. Further investigation on the effects of the three MAPK pathways on Nrf2 transactivation domain activity demonstrated that both ERK and JNK signaling pathways stimulated the Gal4-Nrf2-(1-370) transactivation activity while the p38 pathway played a negative role. Site-directed mutagenesis studies on potential MAPK phosphorylation sites of Gal4-Nrf2-(113-251) showed no significant effect on its basal transactivation activity or the fold of induction by Raf. Interestingly, the nuclear transcription coactivator CREB-binding protein (CBP), which can bind to Nrf2 transactivation domain and can be activated by ERK cascade, showed synergistic stimulation with Raf on the transactivation activities of both the chimera Gal4-Nrf2-(1-370) and the full-length Nrf2. Taken together, this study clearly demonstrated that different segments of Nrf2 transactivation domain have different transactivation potential and different MAPKs have differential effects on Nrf2 transcriptional activity. It also suggested that the up-regulation of Nrf2 transactivation domain activity by upstream MAPKs such as Raf may not be mediated by direct phosphorylation of the Nrf2 transactivation domain, but rather by regulation of the transcriptional activity of coactivator CBP.
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MAPK Signaling Up-regulates the Activity of Hypoxia-inducible Factors by Its Effects on p300
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The Keap1 BTB/POZ dimerization function is required to sequester Nrf2 in cytoplasm
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Cited by
The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress
Nguyen, T., Nioi, P., & Pickett, C. B. (2009) · Journal of Biological Chemistry · doi:10.1074/jbc.r900010200 · Cited by 2,655
Oxidative stress response and Nrf2 signaling in aging
Zhang, H., Davies, K. J. A., & Forman, H. J. (2015) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2015.05.036 · Cited by 804
Stress-activated cap’n’collar transcription factors in aging and human disease
Sykiotis, G. P., & Bohmann, D. (2010) · Science Signaling · doi:10.1126/scisignal.3112re3 · Cited by 776
Molecular mechanisms of Nrf2‐mediated antioxidant response
Li, W., & Kong, A. (2009) · Molecular Carcinogenesis · doi:10.1002/mc.20465 · Cited by 785
Self-citationJeong, C.-B., Kang, H.-M., Lee, M.-C., Kim, D.-H., Han, J., Hwang, D.-S., Souissi, S., Lee, S.-J., Shin, K.-H., Park, H. G., et al. (2017) · Scientific Reports · doi:10.1038/srep41323 · Cited by 585
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Metadata
- Authors
- Shen, G., Hebbar, V., Nair, S., Xu, C., Li, W., Lin, W., Keum, Y.-S., Han, J., Gallo, M. A., & Kong, A.-N. T.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2004-05
- Volume
- 279
- Issue
- 22
- Pages
- 23052-23060
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2135655204
- Cited by
- 194
- References
- 9
- FWCI
- 5.36
- Type
- journal article
- Language
- en
- Open access
- hybrid
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