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• W1968157065 startingPage "L370" @default.
• W1968157065 abstract "Previously, we have reported that endothelial nitric oxide synthase (eNOS) promoter activity is decreased in pulmonary arterial endothelial cells (PAECs) in response to hydrogen peroxide (H 2 O 2 ). Thus the objective of this study was to identify the cis-element(s) and transcription factor(s) responsible for oxidant-mediated downregulation of the eNOS gene. Initial promoter experiments in PAECs treated with H 2 O 2 revealed a significant decrease in activity of a promoter fragment containing 840 bp of upstream sequence of the human eNOS gene fused to a luciferase reporter. However, a promoter construct containing only 640 bp of upstream sequence had a significantly attenuated response to H 2 O 2 challenge. As the 840-bp promoter construct had a putative binding site for the transcription factor activator protein-1 (AP-1) that was lacking in the 640-bp construct, we evaluated the effect of H 2 O 2 on promoter activity after mutation of the AP-1 binding sequence (TGAGTCA at −661 to TGAGTtg in the 840-bp construct). Similar to the results seen with the 640 bp, the AP-1 mutant promoter had a significantly attenuated response to H 2 O 2 . EMSA revealed decreased binding of AP-1 during H 2 O 2 treatment. Supershift analysis indicated that the AP-1 complex consisted of a c-Jun and FosB heterodimer. Furthermore, in vitro EMSA analysis indicated the c-Jun binding was significantly decreased after H 2 O 2 exposure. Using chromatin immunoprecipitation analysis, we demonstrated decreased binding of AP-1 to the eNOS promoter in vivo in response to H 2 O 2 . These data suggest a role of decreased AP-1 binding likely through c-Jun in the H 2 O 2 -mediated decrease in eNOS promoter activity." @default.
• W1968157065 created "2016-06-24" @default.
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• W1968157065 date "2008-08-01" @default.
• W1968157065 modified "2023-10-12" @default.
• W1968157065 title "Hydrogen peroxide decreases endothelial nitric oxide synthase promoter activity through the inhibition of AP-1 activity" @default.
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• W1968157065 doi "https://doi.org/10.1152/ajplung.90205.2008" @default.
• W1968157065 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/2519847" @default.
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