Original ContributionOxidant stress stimulates expression of the human peroxiredoxin 6 gene by a transcriptional mechanism involving an antioxidant response element
Section snippets
Chemicals and reagents
Minimal essential medium (MEM) was purchased from Life Technologies (Grand Island, NY, USA), OptiMEM was from Invitrogen (Carlsbad, CA, USA), and H2O2 and tert-butylhydroquinone (t-BHQ) were from (Sigma). pSEAP2 alkaline phosphatase reporter vectors and an alkaline phosphatase activity assay kit were from BD Bioscience (San Jose, CA, USA); competent cells, restriction enzymes, and the Beta-Glo assay system were from Promega (Madison, WI, USA). All chemicals used were at least analytical grade.
Cell culture
Expression of Prdx6 and Nrf2 in lung epithelial A549 cells
A549 cells were examined to determine whether Prdx6 and the transcription factor Nrf2 are expressed in this cell line. RNA was detected using RT PCR and clearly showed expression of both Prdx6 and Nrf2 (Fig. 1A). Prdx6 protein expression was demonstrated by Western blot, using recombinant Prdx6 as a control (Fig. 1B).
Prdx6 mRNA levels and promoter activity of Prdx6 are increased by H2O2
A549 cells were exposed to H2O2 at concentrations ranging from 50 to 1000 μM for 12 h. There was a concentration-dependent increase in Prdx6 mRNA expression with H2O2 treatment as
Discussion
In contrast to the other peroxiredoxins, Prdx6 contains a single conserved cysteine and utilizes GSH as the redox cofactor to reduce lipid hydroperoxides [9]. Unlike the GSH peroxidases, Prdx6 is not a selenoprotein but uses cysteine as its catalytic center. In view of its antioxidant properties, it is not surprising that oxidative stress induced Prdx6 expression [22]. However, the mechanism of such up-regulation has not been addressed previously.
In this study, we examined the human Prdx6
Acknowledgments
We are grateful to Dr. Anil K. Jaiswal (University of Maryland School of Medicine) for kindly providing the cDNAs encoding Nrf2 and Nrf3, Dr. Shyam Biswal (Johns Hopkins University) for the gifts of siRNA, and Drs. Tatyana Milovanova and Jonni Moore for assistance with cell viability studies. This work was supported by HL P01-75587. I.C. was supported as an NRSA Postdoctoral Fellow by HL T32-07748. This work has been presented in part at the Experimental Biology Meetings of 2005, 2006, and 2007.
References (42)
- et al.
Biological significance of phospholipid hydroperoxide glutathione peroxidase (PHGPx, GPx4) in mammalian cells
Free Radic. Biol. Med.
(2003) - et al.
Cloning, sequencing, and mutation of thiol-specific antioxidant gene of Saccharomyces cerevisiae
J. Biol. Chem.
(1993) - et al.
Mammalian peroxiredoxin isoforms can reduce hydrogen peroxide generated in response to growth factors and tumor necrosis factor-alpha
J. Biol. Chem.
(1998) - et al.
Characterization of a mammalian peroxiredoxin that contains one conserved cysteine
J. Biol. Chem.
(1998) - et al.
Phospholipid hydroperoxides are substrates for non-selenium glutathione peroxidase
J. Biol. Chem.
(1999) - et al.
1-Cys peroxiredoxin, a bifunctional enzyme with glutathione peroxidase and phospholipase A2 activities
J. Biol. Chem.
(2000) - et al.
Structure and phospholipase function of peroxiredoxin 6: identification of the catalytic triad and its role in phospholipid substrate binding
J. Lipid Res.
(2007) - et al.
1-Cys peroxiredoxin knock-out mice express mRNA but not protein for a highly related intronless gene
FEBS Lett.
(2003) - et al.
An antisense oligonucleotide to 1-cys peroxiredoxin causes lipid peroxidation and apoptosis in lung epithelial cells
J. Biol. Chem.
(2002) - et al.
Mice with targeted mutation of peroxiredoxin 6 develop normally but are susceptible to oxidative stress
J. Biol. Chem.
(2003)
Lung injury and mortality with hyperoxia are increased in peroxiredoxin 6 gene-targeted mice
Free Radic. Biol. Med.
NADPH oxidase and ERK signaling regulates hyperoxia-induced Nrf2–ARE transcriptional response in pulmonary epithelial cells
J. Biol. Chem.
γ-Glutamyl transpeptidase is induced by 4-hydroxynonenal via EpRE/Nrf2 signaling in rat epithelial type II cells
Free Radic. Biol. Med.
Nrf2 signaling in coordinated activation of antioxidant gene expression
Free Radic. Biol. Med.
Nuclear import and export signals in control of Nrf2
J. Biol. Chem.
Nrf3 negatively regulates antioxidant-response element-mediated expression and antioxidant induction of NAD(P)H:quinone oxidoreductase1 gene
J. Biol. Chem.
Nitric oxide donors inhibit luciferase expression in a promoter-independent fashion
J. Biol. Chem.
Investigating transcriptional regulation of Prdx6 in mouse liver cells
Free Radic. Biol. Med.
Molecular cloning and functional characterization of a new Cap'n' collar family transcription factor Nrf3
J. Biol. Chem.
Hydrogen peroxide and tumour necrosis factor-alpha induce NF-kappaB-DNA binding in primary human T lymphocytes in addition to T cell lines
Free Radic. Res.
Tolerance of rats to hyperoxia: lung antioxidant enzyme gene expression
J. Clin. Invest.
Cited by (122)
Transcription factor NFE2L3 promotes the proliferation of esophageal squamous cell carcinoma cells and causes radiotherapy resistance by regulating IL-6: NFE2L3 causes radiotherapy resistance in ESCC
2022, Computer Methods and Programs in BiomedicineCitation Excerpt :NFE2L3 belongs to the family of cap'n’collar (CNC), and it is a bZIP (basic-region leucine zipper) transcription factor, along with NFE2L1 and NFE2L2 [10]. Previous studies have indicated that NFE2L3 is involved in oxidative stress and plays a part in the Wnt signaling pathway [11–13]. Recent reports show that NFE2L3 is expressed in a large number of tumors [14], with cell proliferation and invasion promoted by its overexpression, such as in thyroid cancer cells [15], liver cancer [16] and colorectal cancer [17].
Mechanisms underlying Nrf2 nuclear translocation by non-lethal levels of hydrogen peroxide: p38 MAPK-dependent neutral sphingomyelinase2 membrane trafficking and ceramide/PKCζ/CK2 signaling: Mechanism of Nrf2 nuclear translocation by H<inf>2</inf>O<inf>2</inf>
2022, Free Radical Biology and MedicineCitation Excerpt :Previous studies have revealed that oxidative stress induced by supplementing H2O2 to culture media triggers activation of redox sensitive transcription factors such as nuclear factor-E2-related factor 2 (Nrf2), activator protein-1 (AP-1) and nuclear factor κB (NF-κB) dependent on H2O2 concentrations and cell type, to counteract H2O2 and H2O2-induced oxidative damage (reviewed in [24,25]). Table 1 summarizes the activation of these transcription factors by H2O2 in different cell types [26–58]. Nrf2 upregulates expression of genes important for detoxification and defense against oxidative damage [35,59].
NPM1 promotes cell proliferation by targeting PRDX6 in colorectal cancer
2022, International Journal of Biochemistry and Cell BiologyCitation Excerpt :Previous research has shown that SP1-mediated upregulation of PRDX6 can reduce oxidative stress and ferroptosis, thereby preventing podocyte injury in diabetic nephropathy (Zhang et al., 2021). Chowdhury et al. also demonstrated that the binding level of Nrf2 to the PRDX6 promoter was significantly increased after H2O2 treatment (Chowdhury et al., 2009). Interestingly, our findings indicate that NPM1 promotes CRC tumor growth by increasing GPX and iPLA2 activity of PRDX6 (Fig. 4C-D).
Age-related alteration in HNE elimination enzymes
2021, Archives of Biochemistry and BiophysicsDifferential and overlapping targets of the transcriptional regulators NRF1, NRF2, and NRF3 in human cells
2019, Journal of Biological Chemistry