Biochemical and Biophysical Research Communications
The AP-1 site is essential for the promoter activity of NOX1/NADPH oxidase, a vascular superoxide-producing enzyme: Possible involvement of the ERK1/2-JunB pathway
Section snippets
Materials and methods
Materials. PGF2α was purchased from Nacalai Tesque (Kyoto, Japan). Antibodies against c-Fos, c-Jun, JunB, and JunD were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). PDGF-BB was obtained from PEPROTECH (London, UK). [γ-32P]-ATP and [α-32P]-dCTP were from ICN Biomedicals (Costa Mesa, CA, USA).
Cell culture and luciferase assay. The A7r5 cell line, obtained from American Type Culture Collection (Rockville, MD, USA), was cultured in Dulbecco’s modified Eagle’s medium (DMEM)
The AP-1 site is essential for transcriptional activation of the NOX1 promoter
It was reported previously that the consensus MEF2-binding sequence in the promoter region of the rat NOX1 gene was crucial for the inducible expression of NOX1 [14]. In addition to the MEF2-binding site, a consensus activator protein-1 (AP-1) site, 5′-TGAC/GTCA-3′, was found at −98/−92 in the 5′-flanking region (Fig. 1A). To address whether this AP-1 site is responsible for the transcriptional activation, a series of mutants of the NOX1 promoter-luciferase chimera plasmids were constructed. As
Discussion
The major lines of evidence provided by this study are that: (1) the promoter region of the rat NOX1 gene contained a consensus AP-1 site that confers responsiveness to PGF2α; (2) stimulation with PGF2α or PDGF enhanced the binding of JunB to its consensus binding site in the NOX1 promoter; (3) a MEK inhibitor, PD98059, which inhibits the expression of NOX1, suppressed PGF2α- as well as PDGF-induced expression of JunB. Based on these findings and those of our earlier studies [11], [12], [13],
Acknowledgments
We are grateful to Dr. T. Nishinaka, Osaka Ohtani University and Dr. N. Arakawa, Yokohama City University, for valuable discussions and advice.
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2018, Journal of Steroid Biochemistry and Molecular BiologyCitation Excerpt :Indeed, G36 profoundly and selectively reduced expression of Nox1, but not that of Nox2 or Nox4, findings that could be recapitulated in VSMC, intact arteries and myocardium by genetic deletion of GPER (Fig. 1) [10]. Interestingly, it has been proposed that upregulation of Nox1 gene transcription in VSMC is induced by epidermal growth factor receptor (EGFR)-dependent, PI3K- or ERK1/2-mediated activation of the MEF2-binding site and the AP-1 site in the Nox1 promoter region [49–51]. Of note, EGFR transactivation, PI3K and ERK1/2 are signaling pathways typically activated by GPER [22], suggesting potential mechanisms through which GPER regulates Nox1 transcription.
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2016, Pharmacological ResearchCitation Excerpt :However, the role of NOX-1 in atherogenesis remains controversial with NOX-1 being undetected or very low in atherosclerotic rabbit [89] or human lesions [90,91], although NOX-1 overexpression has been described in plaques from patients with cardiovascular events or established diabetes mellitus [92]. NOX-1 promoter has different binding sites for transcription factors [75] including a member of CREB/ATF family [79], AP-1 [93], NF-κB [94] or Janus kinase/Signal transducers and activators of transcription (JAK/STAT) [80]. Most of these studies have evaluated transcriptional regulation of NOX-1.
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2015, Redox BiologyCitation Excerpt :Direct DNA-AP-1 interaction was detected in the promoter of human Nox5 gene in IFNγ-exposed vascular SMCs [51]. A similar AP-1-dependent transcriptional regulatory mechanism has been indicated for the rat Nox1 gene [52]. These data demonstrate that AP-1 is an important regulator of Nox expression and function in various pathological states.
PKCδ mediates paraquat-induced Nox1 expression in dopaminergic neurons
2013, Biochemical and Biophysical Research CommunicationsCitation Excerpt :Miller and collaborators [20] showed that PQ toxicity on microglia cells involves Nox-mediated ROS increase which is regulated by PKCδ. In human neurons, it was reported that PKCδ can increase DNA binding activity of redox-sensitive transcription factor AP-1 [31] which is known to be involved in the regulation of Nox enzymes [32,33]. This could be suggestive of a mechanism by which PKCδ regulates Nox1 transcriptional regulation.
The effect of oxidative stress upon the intestinal epithelial uptake of butyrate
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These authors contributed equally to this work.